Biology 209 Lab Manual

Biology for Allied Health
BI 209L
Anatomy and Physiology Systems
Laboratory Manual
Portland State University

Spring
Dr. Catherine Palmer
Table of Contents
Laboratory Syllabus ............................................................................................................................. 5
Participation/Attendance .................................................................................................................. 5
Grading ............................................................................................................................................. 5
Total Points ...................................................................................................................................... 5
Pre-Lab (9 1 lowest = 8) labs X 10 pt each ............................................................................... 5
Lab/Post-lab Homework (10-1 lowest=9) labs X 20 pts each ..................................................... 5
Quizzes (9-1 lowest=8) quizzes X 10 pts each ............................................................................ 5
Pre-lab Assignments ......................................................................................................................... 5
Post-lab/Homework .......................................................................................................................... 5
Quizzes ............................................................................................................................................. 6
Grading Rubric for Pre- and Post-lab Assignments ......................................................................... 6
Laboratory Rules .............................................................................................................................. 6
Laboratory Safety ............................................................................................................................. 7
Disabilities ........................................................................................................................................ 7
Make-ups .......................................................................................................................................... 7
Laboratory Schedule ........................................................................................................................ 8
Important information: Please fill out on Day 1 .............................................................................. 9
Lab 1 ................................................................................................................................................... 10
Homeostasis ....................................................................................................................................... 10
Background .................................................................................................................................... 10
Exercise 1 ....................................................................................................................................... 10
Answer Sheet .................................................................................................................................. 13
Table 1.1 Effect of HCL and NaOH on Water and Other Substances ........................................... 14
Graphs 1 and 2 ................................................................................................................................ 14
Post-lab Homework 1 ..................................................................................................................... 15
Pre-lab Questions Lab 2 ................................................................................................................. 16
Lab 2 ................................................................................................................................................... 17
Circulation .......................................................................................................................................... 17
Background .................................................................................................................................... 17
Exercise 1 Heartbeat and Pulse ..................................................................................................... 18
Exercise 2 Blood Pressure ............................................................................................................. 19
Exercise 3 Blood ........................................................................................................................... 22
Answer Sheet .................................................................................................................................. 24
Table 2.1 Measuring Pulse Rate .................................................................................................... 25
Table 2.2 Height and Blood Pressure ............................................................................................. 25
Graph 1 ........................................................................................................................................... 25
Lab 3 ................................................................................................................................................... 28
Tissues and the Integumentary System .............................................................................................. 28
Background .................................................................................................................................... 28
Exercise 1 Epithelial Tissues.......................................................................................................... 30
Exercise 2 Connective Tissues ...................................................................................................... 32
Answer Sheet .................................................................................................................................. 35

Lab 4 ................................................................................................................................................... 40
Nutrition and Digestion ...................................................................................................................... 40
Background .................................................................................................................................... 40
Exercise 1 Digestion of Proteins ................................................................................................... 41
Exercise 2 Digestion of Carbohydrates ......................................................................................... 42
Exercise 3 Digestion of Fats .......................................................................................................... 43
Answer Sheet .................................................................................................................................. 45
Table 4.1 Digestion of Macromolecules ....................................................................................... 47
Lab 5 ................................................................................................................................................... 50
The Urinary System ........................................................................................................................... 50
Background .................................................................................................................................... 50
Exercise 1 Kidney Filtration ......................................................................................................... 50
Exercise 2 Urinalysis ..................................................................................................................... 54
Answer Sheet .................................................................................................................................. 57
Table 5.1 Presence/Absence of Glucose, Starch and Chloride Testing ........................................ 58
Table 5.1 Presence/Absence of Glucose, Starch and Chloride Testing ........................................ 59
Table 5.2 Three sample urinalysis ................................................................................................. 59
Lab 6 ................................................................................................................................................... 62
The Immune System ........................................................................................................................... 62
Background .................................................................................................................................... 62
Exercise 1 Immunity ...................................................................................................................... 62
Exercise 2: Disease Transmission .................................................................................................. 66
Exercise 3: Lymphatic System ....................................................................................................... 67
Answer Sheet .................................................................................................................................. 71
Table 6.1 Microorganisms and Disease Lab Data Sheet ................................................................ 72
Table 7.1 Comparison of Hormonal Effects on Different Organs ................................................. 75
Lab 7 ................................................................................................................................................... 76
The Endocrine and Nervous Systems ................................................................................................. 76
Background .................................................................................................................................... 76
Exercise 1 The Nervous System.................................................................................................... 76
Exercise 2 The Endocrine System ................................................................................................. 79
Answer Sheet .................................................................................................................................. 89
Figure 7.1 Unlabeled Neuron ........................................................................................................ 91
Lab 8 ................................................................................................................................................... 94
The Senses .......................................................................................................................................... 94
Background .................................................................................................................................... 94
Exercise 1 The Ears ...................................................................................................................... 94
Exercise 2 The Eyes ...................................................................................................................... 96

Exercise 3 Touch ......................................................................................................................... 102
Exercise 4 Smell .......................................................................................................................... 103
Exercise 5 Taste .......................................................................................................................... 104
Exercise 6 Reflexes ..................................................................................................................... 105
Answer Sheet ................................................................................................................................ 106
Table 8.1 Balance ......................................................................................................................... 108
Table 8.2 Visual acuity................................................................................................................ 109
Table 8.3 Touch Receptors........................................................................................................... 109
Table 8.4 Olfactory Adaptation Data ........................................................................................... 109
Table 8.5 Taste Modalities Data ................................................................................................. 109
Post-lab Homework 8 ................................................................................................................... 110
Pre-lab Questions Lab 9 ............................................................................................................... 111
Lab 9 ................................................................................................................................................. 112
Muscles and Skeleton ....................................................................................................................... 112
Background .................................................................................................................................. 112
Exercise 1 Muscle Tissue ............................................................................................................ 113
Exercise 2 Bone ........................................................................................................................... 116
Answer Sheet ................................................................................................................................ 118
Table 9.1 Length of Glycerinated Skeletal Muscle Following Exposure to Different Solutions 119
Table 9.2 Identification of Unknown Bones ............................................................................... 119
Post lab 9 ...................................................................................................................................... 120
Pre-lab Week 10 ........................................................................................................................... 121
Lab 10 ............................................................................................................................................... 122
Reproduction and Development ....................................................................................................... 122
Background .................................................................................................................................. 122
Exercise 1 Hyperactivation of Bull Sperm Using Caffeine ........................................................ 122
Exercise 2 Development .............................................................................................................. 124
Answer Sheet ................................................................................................................................ 127
Table 10.1 Effects of Caffeine on Sperm .................................................................................... 128
Laboratory Syllabus
You should print and bring a copy of the lab instructions to each weeks lab, but you also will be
able to reference it during class from the classroom computers. Please note that you will not be able
to print the lab from the classroom printer.
Participation/Attendance
Attendance and participation are mandatory. You are required to attend each lab, and to arrive for
all of your labs on time. There are no opportunities to make-up a missed lab, so please be here and
be prepared each week. You are responsible for cleaning your tables with a cleaning solution at the
end of each class and for keeping your work area in tidy condition. In order to earn participation
points you must actively participate in lab and be present for the duration of the lab period. Points
will be deducted if your table/supplies are left dirty or disorganized.
Grading
Lab work makes up 35% of the total grade in Bi209. The points in lab add up to 332, which will be
scaled at the end of the term to be 35% of your points in the class.
Total Points
Pre-Lab (9 1 lowest = 8) labs X 10 pt each
80
In-lab Homework (10-1 lowest=9) labs X 12 pts each
108
Post-lab Homework (9-1 lowest=8) labs X 8 pts each
64
Quizzes (9-1 lowest=8) quizzes X 10 pts each
80
332
Pre-lab Assignments
You will be required to answer pre-lab questions prior to class. The pre-lab assignment must be
handed in as you walk into the lab room each day, beginning the second week of class. Each prelab is worth 10 points. The lowest pre-lab assignment is automatically dropped. 80 points total.
In-lab and Post-lab Homework
If you do not show up to a lab, you may not turn in post-lab homework for that lab.
You must complete each lab, and answer all questions to receive full credit. The lab homework will
consist of two parts: one component that should be shown to your TA at the end of each weeks lab
period (12 points), and post-lab homework that you will finish outside of lab and submit at the
beginning of class prior to the quiz (8 points). Sign your lab with your name, TA's name and lab
time, in pen, in the upper right corner of the first page. Only homework handed in at the beginning
of class is capable of earning full credit. If you turn it in anytime after the start of lab, it is
considered late. Your assignment will still be accepted (up to a week), but for half credit. (This
means that if it is only 20 minutes late, it will lose half the points, so please get your assignments in
on time). If you are 30+ minutes late to a lab, you cannot stay to complete the lab - YOU WILL BE
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ASKED TO LEAVE (no exceptions). Assignments more than one week late will not be accepted.
No late assignments will be accepted the last week of lab. There are 10 labs, worth 20 points each.
The lowest lab homework score is automatically dropped, and there is no post lab assignment on the
last week of class. 172 points total.
Quizzes
At the beginning of each lab, you will be given ~10 minutes to complete a quiz. The quiz will
include 8 questions from the previous weeks lab, as well as 2 basic questions about the current
days lab activities. This means you must read the lab manual before you arrive to class. The
questions will be multiple choice, short answer and/or fill in the blank. Each question will be worth
one point. If you are more than 10 minutes late, you will not be permitted to take the quiz. There
will be a quiz every week except the first week, each of which will be worth 10 points. Your lowest
quiz score will automatically be dropped. 80 points total.
Grading Rubric for Post-lab Assignments
Content
70 Percent
All key elements of the assignment are covered in a substantive way.
Questions are answered correctly and fully, demonstrating an appropriate and welldeveloped understanding of the material
Organization/Development
15 Percent
All responses are written in complete sentences.
The assignment is typed
Mechanics
15 Percent
Rules of grammar, usage, and punctuation are followed; spelling is correct.
Laboratory Rules
ALWAYS READ THE LAB EXERCISE BEFORE COMING TO CLASS
Turn your phone off for the duration of the lab.
Arrive on time and prepared for 2 hours of work.
The computers are for Bi 209-related work only, e.g. data collection, investigation of topics
related to the lab exercise or lectures.
Students are expected to complete all assignments independently. Cheating and/or
plagiarism will not be tolerated. Please consult PSUs Student Conduct Code for more
information: http://www.pdx.edu/dos/conduct.html. Cheating and any other academic
dishonesty will result in a zero for the assignment, potentially a zero for the entire lab
(TA's decision), and will be reported to the University administration. If you have a
phone in your hand at any point during a quiz you will receive a zero as your lab grade
for the term.
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Leave the lab benches cleaner than you find them. Messes that someone else has to clean up
will result in point deductions.
Laboratory Safety
Please use caution and common sense at all times.
Report any injuries or broken equipment to the TA immediately.
No food (including gum or candy) or drink is allowed in the lab.
No sandals or open shoes of any kind. You will be asked to leave the lab if you are not in
compliance.
Waste containers must be used for all designated substances. Your TA will tell you what can
be disposed of in the sink.
If you choose not to use these safety precautions you will be liable for any injury you may
incur.
Disabilities
If you are a student with a documented disability and registered with the Disability Resource
Center, please notify Dr. Palmer immediately to facilitate arranging academic accommodations.
Make-ups
There will be no opportunities to make up a missed lab or quiz. You are able to drop your lowest
scores, so this should account for any need to make up a lab.
Laboratory Schedule
Lab #
Topic
Quiz or Exam
Due today
Homeostasis
Circulation
Quiz #1
Week 2 pre-lab Week

1 homework
Tissues and the

Integumentary System
Quiz #2
Week 3 pre-lab Week

2 homework
Nutrition and Digestion
Quiz #3
Week 4 pre-lab Week

3 homework
Urinary System
Quiz #4
Week 5 pre-lab Week

4 homework
Immune System
Quiz #5
Week 6 pre-lab Week

5 homework
Endocrine and Nervous

Systems
Quiz #6
Week 7 pre-lab Week

6 homework
The Senses
Quiz #7
Week 8 pre-lab Week

7 homework
Muscles and Skeletal

System
Quiz #8
Week 9 pre-lab
Week 8 homework
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Reproduction and
Development
Quiz #9
Week 10 pre-lab
Week 9 homework
Important information: Please fill out on Day 1

Your TAs name: ________________________
TA contact information: ________________________
Your labmates name(s) and email address(es):
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
Lab 1
Homeostasis
Background
The cell is the basic unit of your body, forming its tissues, organs, and organ systems. Each cell in
your body is specifically adapted to perform specialized functions. However, the cell can only
function under very specific environmental conditions. Your body is made up of trillions of cells,
each of which has its own preferred environmental requirements, yet your body as a whole can
function optimally under a single specific set of conditions. The body, and essentially all of its cells,
therefore defend and maintain this relatively constant environment. Homeostasis, this ability to
regulate and maintain a steady state, is a feature that defines all living organisms. Some examples
of homeostasis in your own body include the maintenance of constant body temperature (98F),
keeping your blood pH within a certain range, maintaining levels of glucose in your blood, and
keeping and balancing food/water intake with waste removal.
Since the body strives to maintain a constant state, any deviations can result in non-optimum
performance of tissues, organs, organ systems and eventually the organism. This non-optimum
performance of body structures is part of what we call disease. Therefore, the inability of the body
to maintain any of the many internal variables results in disease. In a healthy individual,
homeostasis can be temporarily overcome through appropriate autonomic adjustments in the
physiological systems. For example, as you exercise, your cardiac and respiratory rates are
increased in response to increasing levels of blood CO2. After exercise, your body must then
readjust the cardiac and respiratory rates to reflect the decrease in metabolic demands. If this
readjustment is not accomplished, your elevated breathing rate can lead to respiratory alkalosis
(increased pH), loss of consciousness, and possible permanent nerve damage. The failure of the
body to maintain homeostasis, resulting in disease, can occur for a variety of reasons. Some of these
include, infection (bacterial, viral), genetic disposition, immune disorders, trauma, aging, nutritional
factors, etc.
Exercise 1
For the lab exercises today, neatly answer the questions on the answer sheet provided using
complete sentences. It is best to use pencil.
One factor that affects the internal stability of plants and animals is the relative concentration of
hydrogen ions (H+) and hydroxide ions (OH-). High levels of H+ ions results in an acidic
environment, and high levels of OH- ions produces a basic environment. Metabolic activities of
living tissues produce an abundance these ions, yet life depends on maintaining a pH range that is
within certain limits for each tissue or system. Using pH paper, you will be looking at the responses
of several substances to the addition of an acid and a base. During this exercise, consider how
organisms survive and function despite metabolic activities that tend to shift pH toward either acidic
or basic ends of the pH scale.
Question 1. Develop a hypothesis that addresses how you think water will respond to the
addition of HCl (an acid) and NaOH (a base).
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Question 2. Develop a hypothesis that addresses how you think the biological material you
were assigned will respond to the addition of HCl (an acid) and NaOH (a base).
MATERIALS
tap water
2 small beakers
Pipette/dropper
pH paper
0.1M HCl
0.1M NaOH
Potato homogenate
Egg white (diluted 1:5 with H20)
Whole milk
Graph paper
1) Work as a table. Pour 25 mL of tap water into a beaker.
2) Record the initial pH of the water using a small strip of pH paper.
3) Add 0.1M HCl one drop at a time. Gently swirl the mixture after each drop is added. After
adding 5 drops, use a strip of the pH paper to record the pH in Table 1.1.
4) Repeat this procedure five more times so that you have a total of 30 drops in the water. Measure
the pH after every five drops and record the pH in Table 1.1.
5) Properly dispose of your liquid and rinse the beaker really well.
6) Pour another 25 mL of water into the beaker.
7) Measure the pH using pH paper and record the initial pH in Table 1.1.
8) Add 0.1M NaOH one drop at a time just like above with the HCl.
9) Record your data for each 5 drops added until 30 drops have been added in total.
10) Properly dispose of your liquid and rinse the beaker really well.
Your table will be assigned one of the following: potato homogenate, egg whites, or milk.
11) Repeat the above procedure, replacing water with one of the material youve been assigned:
potato homogenate, egg white, or milk.
12) Record your data for both the acid and the base in Table 1.1.
13) Obtain class data and average pH values after each set of 5 drops was added to the water and
record these average values in Table 1.1.
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14) Record all class data (averages for all materials) in Table 1.1.
Graph 1 and 2: Create two graphs of your class average data: one showing the addition of acid to
each of the liquids (water, potato, egg white, milk) and one showing the addition of base to each of
the liquids.
Use the graphs that you made to answer the following questions.
Question 3. Summarize the effect of the HCl and NaOH on tap water.
Question 4. Compare the change in pH values of water. Which test gave the largest pH
change? Which gave the smallest?
Question 5. How do biological materials react to changes in pH?
Question 6. Compare the change in pH values of biological material. Which test gave the
largest pH change? Which gave the smallest?
Question 7. For both the water and the biological material, does the pH level off after a
while?
Question 8. Briefly compare the pH changes you observed in the tap water with the pH
changes you saw overall in the biological materials (the potato, egg white, and milk), describe
any differences or similarities.
Question 9. Based on what you can see from your data, does your data support your
hypothesis? Why or why not?
Question 10: What does your data suggest about a mechanism living organisms might use for
regulating pH?
Be sure to show your answer sheet, tables and graphs to your TA before you leave today.
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Answer Sheet
Please print this before class and be sure to show it to your TA before you leave the
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
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Table 1.1 Effect of HCL and NaOH on Water and Other Substances
pH
Water
Potato
Milk
Your
Class
Your
Class
Your
Class
data
Ave
data
Ave
data
Ave
HCl
0 drops
5 drops
10 drops
15 drops
20 drops
25 drops
30 drops
NaOH
0 drops
5 drops
10 drops
15 drops
20 drops
25 drops
30 drops
Graphs 1 and 2
(graph on separate sheets of paper)
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Egg White
Your
Class
data
Ave
Post-lab Homework 1
Post lab questions (submit answers to these to your TA at the start of lab 2):
You may be required to use outside sources (such as your textbook) to answer post-lab
questions.
1. Describe homeostasis (do not plagiarize).
2. Discuss two sources of error in your lab experiments and how they could have affected your
results. Then give recommendations for fixing these sources of error.
3. What mechanisms does your body use to maintain its homeostatic temperature?
4. Briefly describe the process of a fever, including benefits and possible damaging effects. (Refer
to your textbook for assistance).
5. Define and give an example of a positive feedback mechanism in your body.
6. Define and give an example of a negative feedback mechanism in your body.
7. How do positive and negative feedback mechanisms allow you to maintain homeostasis?
8. Describe a disease that results from the bodys inability to maintain homeostasis.
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Pre-lab Questions Lab 2

Answer the questions below. Your answers will be handed in at the beginning of lab 2.
1. The cardiovascular system includes three things. Name them.
a.
b.
c.
2. Blood leaves the heart via vessels called __________________ and returns to the heart via
vessels called _________________.
3. Valves between the atria and ventricles are called what?
4. Name and define four components of blood.
5. How many molecules of oxygen can a single red blood cell carry?
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Lab 2
Circulation
Background
Every one of your trillions of cells has the same basic needs each must take in oxygen and
nutrients, and get rid of carbon dioxide and other metabolic wastes. If these simple needs are not
met, the cell will die. Fortunately, you have an intricate network of vessels that transport fluids
carrying gases, nutrients and wastes to and from the cells in your body. This network of vessels
makes up your circulatory system. More specifically, the circulatory system of a vertebrate such
as yourself is called the cardiovascular system. The cardiovascular system includes the heart, the
blood, and the blood vessels (i.e., arteries, capillaries, and veins), and it connects all of the systems
of your body. The blood vessels are all connected to form one continuous closed system. The heart
provides the force needed to move approximately five liters of blood out to your blood vessels
nearly every minute. The blood carries the nutrients and oxygen to your cells and whisks away the
carbon dioxide and other wastes for disposal.
Do you see how vitally important this system is? When something goes wrong with this system, the
underlying disease is called cardiovascular disease. Cardiovascular disease is the leading cause of
death in the U.S., claiming one life every 35 seconds (2500 Americans a day). What is perhaps
surprising is that most cases of cardiovascular disease are preventable. To keep your circulatory
system healthy and vibrant, dont smoke, cut back on saturated fats (totally eliminate trans fats!),
eat a healthy diet full of fresh foods, exercise, manage stress, and maintain a healthy weight. Most
forms of cardiovascular disease start developing when you are young, but virtually all of the
damage can be reversed with simple lifestyle changes early on.
Blood vessels
The three types of blood vessels include the arteries, veins,
and capillaries. The arteries carry blood away from the heart.
They are the widest blood vessels and have thick, elastic
walls. Arteries become thinner arterioles that lead to
capillary beds. Capillaries are the tiniest of blood vessels,
usually only one cell thick, that feed the tissues. Capillaries
are unlike veins and arteries in that they are very leaky
water, nutrients, oxygen, carbon dioxide, and other metabolic
wastes can diffuse into and/or out of capillaries via diffusion.
From the capillary bed, blood moves into venules, which
lead to larger veins that empty into the heart. Veins always
carry blood toward the heart, so most veins are carrying
blood that is laden with carbon dioxide and other metabolic
wastes. Veins are usually closer to the skin and contain
valves that prevent backflow and maintain flow toward the
heart.
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The Heart
Your heart has four chambers; two
ventricles and two atria. The heart serves
as a pump to drive the flow of blood through
the body. It does so by undergoing a cycle of
contraction and relaxation called the cardiac
cycle. During the cardiac cycle, blood is
pumped into the atria and out of the
ventricles. First, an electrical signal is
generated in the pacemaker cells and is
distributed throughout the heart via an
electrical conduction system. In response to
electrical stimulation, specialized muscles of
the atria contract first, and then the ventricles
undergo contraction. Contraction of the heart
is called systole. Contraction of the atria
sends blood to the ventricles. Once the ventricles fill with blood, the atrioventricular valves
between the atria and ventricles close to prevent blood from flowing back into the atria. When the
AV valves snap shut, a lub sound is heard. When the ventricles contract, the right ventricle sends
blood out to the pulmonary circuit (to the lungs) and the left ventricle sends blood out through the
aorta to the systemic circuit. The semilunar valves of the ventricles close once the blood is
pumped out, and the closing of these valves makes a characteristic dub sound. Together, this
series of closing valves produces the sound of your heart beat lub-dub, lub-dub, lub-dub. The
injection of blood into the arteries by the ventricles undergoing systole generates blood pressure, the
primary driving force for the flow of blood through the body. Following systole, the heart muscles
relax. This relaxation of the heart muscle is called diastole. During this relaxed state, the valves
between the atria and the ventricles open and all four chambers of the heart begin to fill with blood.
Exercise 1 Heartbeat and Pulse
Your heartbeat is a measure of the cardiac cycle of blood flowing into your relaxed heart, followed
by the contraction of the atria moving the blood into the ventricles, and the subsequent contraction
of the ventricles moving the blood into the pulmonary and systemic circuits. As the blood surges
into the arteries during a heart beat, each artery stretches and bulges. This brief bulge of the artery
is called a pulse. The number of pulses per minute, your heart rate, determines how fast your heart
is beating.
Materials
- Stethoscope
-Timer or stopwatch
1) Obtain a stethoscope and wipe the earpieces with an alcohol wipe to make sure they are clean.
Sit comfortably in a chair, do not cross your legs.
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2) Put the earpieces in your ears. Notice that the earpieces are angled. The earpieces should be
angled in a forward direction when placed into the ears. Can you hear anything? Place either side
of the stethoscope to your chest. Do you hear anything? If not, try the other side. Listen for the
lub-dub of your heart.
Question 1: What produces the sound of the first part of your heartbeat? What produces the
sound of the second part of your heartbeat? Which part of the heartbeat is longer? Which
part is louder?
3) Set the stethoscope aside and grab a timer or stopwatch. Remain seated.
4) To feel the pulse, find the artery in your partner's wrist. Place the tips of the first two fingers of
one hand on the palm side of your partner's wrist, over toward the thumb side of his or her wrist.
You may need to press quite firmly in order to feel the pulse of blood which each heart beat sends
through the artery. Don't use your thumb to feel the pulse in the wrist, because your thumb has a
pulse of its own.
5) To measure heart rate, count the number of pulses in 30 seconds. It helps to compress the artery
firmly as you begin, and then immediately ease up on the pressure slightly. Multiply that number
by 2, and you will have the number of heart beats per minute. The average is 70-76 beats per
minute in resting state.
Question 2. What do you notice about the regularity (or perhaps irregularity) of the pulse?
6) Measure the pulse using the conditions shown in Table 2.1. Record your results in Table 2.1.
Question 3. How does your pulse rate vary from activity to activity? Why does the pulse rate
change?
Question 4. Describe what is happening in your heart and in your blood vessels when your
pulse rate increases.
Question 5. Why do you think it is necessary for your pulse rate to increase when you
increase activity?
Exercise 2 Blood Pressure

Materials
-measuring tape
-sphygmomanometer
-graph paper
Blood pressure is the pressure the blood exerts against the walls of the blood vessels, and is usually
measured in the arteries. Blood pressure in the arteries oscillates during the cardiac cycle. Normal
systolic blood pressure -the pressure in the arteries during ventricular contraction - is ~120
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mmHg. Normal diastolic blood pressure - pressure in the arteries during ventricular relaxation - is
somewhat lower at ~80 mmHg. Therefore, normal blood pressure is said to be 120/80
(systolic/diastolic). The difference in pressure between systole and diastole is called the pulse
pressure, which is a useful diagnostic measure for cardiovascular health. Blood pressure can
change based on activity levels and on body position. For example, when a person is standing,
blood will tend to be drawn into the extremities (particularly the legs) with the force of gravity.
Thus, the heart will need to pump harder in order to recover blood and to deliver blood to the brain
against the force of gravity, and blood pressure will become elevated. In contrast, if a person is
reclining, blood tends to pool in the abdomen and thorax, and the effects of gravity become less.
When reclining, the heart does not need to pump blood as rigorously to ensure adequate circulation,
and the blood pressure will lower.
The chart below shows blood pressure classifications. This measurement varies from individual to
individual and can be dependent on many factors (see chart). Also, blood pressure measurements
change throughout the day. Hypertension is high blood pressure and is known as the silent killer
because it can cause heart attacks, strokes, and even heart failure, with no prior symptoms at all.
Blood Pressure Ranges
Some factors that affect
blood pressure
-age
-weight
-heredity
-time of day
-physical activity
-emotional stress
-certain foods
-medications
Rating
Systolic
Diastolic
optimal
<120
<80
normal
<130
<85
high normal
130-139
85-89
hypertension
stage 1
140-159
90-99
hypertension
stage 2
160-179
100-109
hypertension
stage 3
>179
>109
Question 6: How do you think blood pressure would differ (if at all) in the following
situations and why: Standing versus lying down? Walking versus running?
If gravity affects blood pressure, then would you expect people of different heights to have different
blood pressure? This is the question we are going to investigate in this lab exercise.
Question 7: Write your hypothesis and prediction based on todays lab question (on height).
You will use a sphygmomanometer, a blood pressure cuff, to measure blood pressure. The
instrument consists of an inflatable cuff with an attached pressure gauge. The cuff is placed on the
arm above the elbow and then inflated to a pressure high enough to prevent circulation to the
forearm. As pressure in the cuff is gradually released, the examiner listens with a stethoscope for
characteristic sounds of blood flow into the forearm. These sounds are called the sounds of
Korotkoff. The pressure at which first sound of turbulence is detected is recorded as the systolic
pressure. As more pressure is released from the cuff, the turbulence increases, as does the loudness
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of the sounds. More and more pressure is released until no turbulence is detected. The pressure at
which sounds disappear is recorded as the diastolic pressure.
1) Choose a lab member who will have their blood pressure measured.
2) Measure the height of the student and record the results (in inches) in Table 2.2.
3) Clean stethoscope ear pieces with alcohol wipes.
4) Have student sit in a chair, feet planted on the floor (no leg crossing!).
5) Snugly place sphygmomanometer two to three finger-widths above crease in the arm.
6) Have the student adjust the height of his or
her arm crease so that it is level with his/her
heart.
7) Place stethoscope directly under bottom of
cuff (but not in the arm crease) and put ear
pieces in.
8) With the valve closed, begin to increase
pressure in cuff with bulb until there is no blood
flow (around 180mm Hg).
9) Allow pressure to slowly drop by releasing the valve.
10) When you begin to hear blood flow, read the measurement (systolic pressure) while still slowly
releasing pressure. Say the number out loud to your lab partner(s) so that it can be recorded in
Table 2.2.
11) The blood flow will keep getting louder and right at the moment you dont hear a sound read
the measurement (diastolic pressure). Say the number out loud to your lab partner(s) so that it can
be recorded in Table 2.2. Share your results with the class.
Test your Accuracy Using the Blood Pressure Sensor with a Computer
1) Connect the Blood Pressure Sensor to Channel 1 on the LabPro LabQuest, LabQuest 2, LabQuest
Mini, or to a Go! Link connected to a computer. Attach the rubber hose from the cuff to the
connector on the sensor.
2) Wrap the cuff firmly around your partner's arm, just less than one inch above the elbow. The two
rubber hoses from the cuff should be positioned over the bicep muscle (brachial artery) and not
under the arm. Important: the person having his or her blood pressure measured must remain still
during data collection - no movement of the arm or hand during measurements.
3) Start Logger Pro or Logger Lite on the computer.
21
4) Logger Pro or Logger Lite will identify the Blood Pressure Sensor and load the proper
calibration. You are now ready to collect data.
5) Click 'Collect' to begin collecting data.
6) Quickly and repeatedly squeeze the bulb to inflate the cuff on your partner's arm. Continue
inflating the cuff to a pressure between 150 and 170 mm Hg. A meter in the data collection
software will display the live pressure readings from the sensor. When the maximum pressure is
reached, set the bulb pump down onto the table. The built-in pressure release valve will slowly
deflate the cuff.
7) After the pressure drops to 50 mm Hg, you may press down on the pressure release valve to
release any air left in the cuff. If the pressure does not reach 50 mm Hg by the time data collection
ends, let your TA know so that they can adjust the pressure release valve according to directions in
the software manual.
Question 8. Were your results the same using the sphygmomanometer manually vs. using the
computer program. What might account for differences?
Graph 1. Using the class data, create a graph of the blood pressure/height data for the class.
Question 9. Does there appear to be a trend in your data? If so, describe it.
Question 10. Was your hypothesis supported or rejected? Provide an explanation of your
results.
Question 11. What are some variables that may have had an impact on your results? What
could you do differently next time to avoid these problems?
Exercise 3 Blood
Blood is a fluid connective tissue composed of
plasma, red blood cells, white blood cells, and
platelets. Plasma is the liquid portion of the
blood and is made up mostly of water, but it also
contains several salts, hormones, proteins, amino
acids, glucose, nutrients, and metabolic wastes.
Serum is blood plasma from which a protein
called fibrinogen is removed. Fibrinogen plays
an important role in blood clotting. Red blood
cells, erythrocytes, are the most numerous type
of cell in your blood.
They are highly
specialized to perform one specific taskthe
transport of oxygen. Red blood cells have lost
most of their organelles, including the nucleus
(and the DNA it normally carries). The loss of
22
organelles increases the volume inside the cell for transporting oxygen which makes red blood cells
super efficient at their task. Oxygen is carried by hemoglobin, an iron-containing pigment (a
protein) that turns red when oxygen binds to it. A single molecule of hemoglobin can carry four
oxygen molecules, and each red blood cell is packed with approximately 250 million molecules of
hemoglobin. Erythrocytes live for only a few months before they wear out and are recycled by your
body. White blood cells, leukocytes, come in many forms and function in your immune system.
These cells patrol your blood and tissues, looking for bacteria, viruses, cancer cells, or any other
invaders. There are many different types of white blood cells, which we will examine in greater
detail when we talk about the immune system. Lastly, platelets (thrombocytes) are fragments of
bone marrow cells that play a vital role in clotting the blood. Platelets last for approximately seven
days in the bloodstream. Bone marrow contains stem cells that can develop into red blood cells,
white blood cells, or platelets.
Materials
-Human blood slides
-Compound microscope
1) Obtain a human blood slide and a compound microscope.
2) Examine the slide of human blood starting with the 4X objective and moving up to the 40X
objective.
Question 12. What color are the majority of cells on your slides?
Question 13. Can you identify any cellular structures within the red blood cells?
Sketch 1. Sketch the red blood cells as they appear in your field of view under 40X
magnification. Label any organelles that may be present.
Question 14. Move around the slide until you locate a cell that is not a red blood cell. What
difference do you notice immediately?
The two most common white blood cells are lymphocytes and neutrophils. The lymphocytes that
youll see in this slide fight infections. Lymphocytes have a large, round nucleus that nearly fills
the cell. Neutrophils ingest infectious bacteria and cellular debris. The nucleus of a neutrophil is
shaped almost like an hourglass.
Sketch 2. Sketch the white blood cell that you see on your slide. Label any organelles that
may be present.
Question 15. Did you sketch a neutrophil or a lymphocyte?
Be sure to show your answers, tables, and sketches to your TA before you leave today.
23
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
Question 11
24
Question 12
Question 13
Question 14
Question 15
Table 2.1 Measuring Pulse Rate
Condition
After sitting still for 5 minutes
Immediately after standing up
After running in place for 30 seconds
Table 2.2 Height and Blood Pressure
Height
Systolic blood
(inches)
pressure
Student 1
Student 2
Student 3
Student 4
Student 5
Student 6
Student 7
Student 8
Student 9
Student 10
Pulse Rate (beats/min)
Diastolic
Graph 1
(use separate sheet of paper)
Sketch 1
Sketch 2
25
Post-lab Homework 2
questions
1) What are the cellular components of blood and their general functions?
2) How does the circulatory system help the body maintain homeostasis?
3) How do capillary beds assist in the function of the cardiovascular system?
4) Why are the ventricles larger than the atria?
5) Explain the flow of blood through the heart and body (include both circuits, the vessels, and
names of the chambers and valves).
6) Explain the difference between the pulmonary and systemic circuits.
7) Describe the difference between systole and diastole (be very specific- check your lecture slides
and book for details).
8) What factors can affect pulse and blood pressure? Why does the pulse and blood pressure
change in response to these factors?
26

1. Define tissue.
2. Describe the four types of animal tissues.
3. If an epithelium consists of only one layer of cells on a basement membrane, it is called
________________________. More than one layer of cells on a basement membrane represents
________________. A tissue with a single layer of thin flattened cells is called _______________,
whereas a tissue with multiple layers of cells having height and width of equal dimension is called
________________.
4. Describe the different types of connective tissue.
5. The integumentary system includes what? Describe the three layers that make up skin. In which
layer are hair follicles, sebaceous glands, arrector pili muscles and sweat glands found?
27
Lab 3
Tissues and the Integumentary System
Background
Your cells are highly dependent upon one another and must cooperate in order to perform complex
tasks and maintain homeostasis within the body. During development, cells become specialized some become skin cells, some become bone cells, some make up the eye, the liver, the kidney, etc.
As cells become specialized, they take on specific functions within the body. Groups of cells that
are similar in structure and function are called tissues. There are four types of tissues within your
body epithelial, connective (or supportive), nervous, and muscular tissue. Each type has
distinct structures and functions. Tissues that work together to perform specific functions are
known as organs. In turn, these organs may exhibit a structural and functional interdependency,
resulting in organ systems. One such system that you will study today is the integumentary system.
Epithelial tissues are made up of cells that are tightly-packed together leaving very little space for
intercellular material or matrix. Epithelial tissues line all the external surfaces and internal cavities
of an animal's body as continuous sheets of cells. Some epithelial tissues found in glands are
specialized for secretion, such as the glandular epithelia that secrete hormones or digestive
enzymes. An important "landmark" in studying epithelial tissues is the basement membrane, a
non-cellular, fibrous layer of material that separates the epithelium from the underlying tissues. The
cells are joined together by different types of cell anchoring or communication structures. All
materials entering or leaving the body must ultimately cross at least one layer of epithelial tissue. In
general, four functions may be attributed to epithelial tissues: (1) protection of the underlying
tissues, (2) absorption, (3) secretion, and (4) reception of sensory stimuli.
Different types of epithelium may be distinguished by their morphological appearance, which in
turn is dependent on the location of the tissue and the particular tasks it must accomplish. If an
epithelium consists of only one layer of cells on a basement membrane, it is called simple
epithelium. More than one layer of cells on a basement membrane represents stratified
epithelium. In addition, the cross-sectional shape of epithelial cells can be used to further classify
these tissues. Thus, thin flattened cells are squamous, cells with height and width of equal
dimension are cuboidal, and rectangular cells with the height greater than the width are columnar.
28
Connective tissue is found throughout the body and is the most abundant and widely distributed
tissue type. Connective tissues are made up of cells that are widely separated so the space in
between them is filled with extracellular matrix. The matrix is secreted by the cells and has two
components, ground substance (mostly glycoproteins and polysaccharides) and fibers (collagen and
elastin). The job of connective tissue is to protect, support, and bind together tissues. Classification
of connective tissues is based upon the arrangement of the extracellular matrix and the substance
that make up the extracellular matrix. Connective tissues include bone, cartilage, ligaments and
tendons (dense connective tissue), blood, adipose (fat) tissue, and loose connective tissue.
Muscular tissues are designed for contraction and relaxation. The two major contractile proteins
are actin, and myosin. Muscular tissues are classified based upon the organ where it occurs, the
histological appearance of the cells, and the mode of enervation. We will examine muscular tissue
in week 9.
The nervous tissue is found primarily in the brain, spinal cord, as well as in ganglia and other
organs of the body. The major type of cell is called neuron. It is designed to receive and transmit
nerve impulses. Some nervous tissues are made up of modified cells such as the receptors in the
skin and sensory organs. We will examine nervous tissue in week 7.
The integumentary system includes the skin, hair,
and nails. The two main layers of the skin include
the epidermis (outermost layer), and the dermis
(main middle layer). Beneath the dermis is the
hypodermis (subcutaneous tissue; innermost
layer). The epidermis is stratified squamous
epithelial tissue consisting of 4-5 distinct layers.
The dermis is connective tissue and it consists of
several structures, including hair follicles,
sebaceous glands, arrector pili muscles and
sweat glands. The hair follicle produces the hair.
The arrector pili muscle is attached to the hair
follicle. When it contracts, it pulls the hair follicle
up and forms a goose bump. The sweat glands
are located throughout the dermis and have a duct
that leads to the outside of the skin. The
hypodermis consists of adipose tissue and is not
considered part of the skin. It is below the skin.
29
The part of the hair that is enclosed in a hair

follicle is called the root; the portion projecting
from the skin is the shaft. The hair is formed by
mitosis of the germinal epithelial cells at the
base of the follicle (the hair bulb). As the
daughter cells are pushed away from the
growing region, they become keratinized and
die, thus the bulk of the hair shaft is dead. A
hair consists of a central region, the medulla,
surrounded by the cortex and then the protective
cuticle. The cuticle is usually composed of
overlapping cuticular scales.
Exercise 1 Epithelial Tissues
Materials
-prepared slide of skin
-Magnifying glass
-microscope slides
-cross section of small intestine (slide)
- prepared slide of a kidney
-prepared slide of a trachea
1. Examine a prepared slide of the skin at low, medium and high magnification. Look for the cells
in the epidermis that are tightly packed together. Focus on an individual cell and describe its shape.
Squamous epithelia are made up of cells that are flat and polygonal. This epithelial tissue is also
called stratified squamous epithelium because there is more than one layer of cells in the tissue.
Stratified epithelia are normally found in places where there is a great deal of wear and tear.
Frequently, the outer surface of these epithelia is sloughed off and replaced by cells below it. In
areas where the tissue is subjected to drying it may be keratinized. In keratinized stratified
squamous epithelium the outermost layer is composed of tightly packed dead cells filled with the
protein keratin. Keratin is the same protein which makes up hair and fingernails. The layer of
keratin-filled cells acts as water-proofing.
Sketch 1. Sketch your observation and label the cells, basement membrane, and nucleus.
Label the area that is outside the body. Label the dermis and epidermis. Label any other
structures that you recognize.
Question 1. What happens to the nuclei as cells migrate from the basement membrane
outward to replace the sloughed surface?
30
Question 2. Using the high magnification image, compare the sizes of the cells nearest to and
farthest from the basement membrane. Is there any difference in size?
Find a hair follicle (hair shafts are visible in the low and mid magnification) and notice that the cells
that make it up are continuous with the epidermis. The follicle cells secrete the protein material
(keratin) that forms hair.
Question 3. What function(s) does hair perform? Compare the relative roles of hair in
humans and other mammals, such as rodents.
Question 4. In what other organs would you expect to find stratified squamous epithelium?
2) Pull out a strand of your hair and examine it with a magnifying glass. You may need to put it on
a piece of white or black paper to make it easier to see.
Question 5: What does the root look like?
Choose one: teardrop/narrow/rounded/pointed/other (describe)
Question 6: What does the tip look like?
Choose one: frayed/smooth/bent/split/other (describe)
Question 7: What color is the shaft? Is the color the same everywhere along the shaft?
3) Place your hair on a slide and view the shaft and root at low, medium, and high power.
Question 8. How would you describe your cuticle scales?
protruding or spiky, or neither?
Are they smooth and flat,
4) Examine a prepared slide of the small intestine. Look for the villi, the finger-like projections into
the lumen. Adjust your magnification to focus on the individual cells.
Question 9. Describe the shape of the cells.
This tissue is also called simple because there is only one layer of cells forming the tissue. The cells
are tall and are called columnar, hence this tissue is called simple columnar epithelium. These large
epithelial cells are specialized for absorption, and are commonly found in areas where there is
considerable wear and tear, such as the digestive tract. Find the goblet cells, modified columnar
cells that secrete mucus among the columnar cells that form the mucosa layer of the small intestine,
and show them to your TA.
Since one of the functions of the intestine is absorption, an adaptation of the free surface of these
epithelial cells is the presence of microvilli (see the drawing on the next page).
Question 10. What do these microvilli accomplish?
31
5) Examine the prepared slide of a kidney. Focus on the cross section of the tubules, then focus on
one cell bordering one tubule.
Question 11. Describe the shape of the cells.
Question 12. How many layers of cells form the tubules?
Question 13. How would you classify this type of tissue?
Sketch 2. Sketch your observations and label the cells.
6) Examine the prepared slide of the trachea. Note that the inner surface or lumen is lined by some
tall and some short cells that have tuft of hair-like structures on their exposed surface. This tuft-like
structure is called cilia, and the continuous beating of the cilia propel dust and other undesirable
substances out of the respiratory organ. If you look towards the basement membrane, you will note
that each cell is resting on it. Although the section may appear stratified, it is actually
pseudostratified. This tissue is called pseudostratified ciliated columnar epithelium. You may
also find goblet cells, modified columnar cells that secrete mucus.
Sketch 3. Sketch your observations and label the columnar cells, basement membrane, cilia,
goblet cells (if present).
Exercise 2 Connective Tissues
As the name implies, these tissues perform connective and supportive functions: giving form to and
anchoring the organs of the body. Unlike epithelium, connective tissues consist of one or more cell
types embedded in a matrix. The matrix consists of widely dispersed fibers embedded in a ground
substance, which may range in form from solid to liquid.
Materials
-compound microscope
-prepared slide of skin
-prepared slide of white fibrous connective tissue
32
-prepared slide of adipose tissue

-prepared slide of cartilage
1. Examine the prepared slide of the skin (the same slide you examined when you studied the
epithelial tissue). Focus on the thick, loose layer beneath the stratified squamous epithelia layer.
This region of the skin called the dermis consists of connective tissue. The dermis is composed of
two layers: (1) the papillary dermis, the thin top layer closest to the epithelium, is composed of
less dense connective tissue (2) the thick underlying layer of the dermis, the reticular dermis, is
composed of dense irregular connective tissue. Note how loose the fibers are in the papillary dermis
and how far apart the cells appear. When the skin is young, it appears taut and smooth because the
cells that secrete the fibers, the fibroblast cells, are plentiful and are actively producing collagen
fibers, the matrix of the connective tissue. As the person ages, the skin becomes wrinkled because
the ability of the fibroblasts to produce collagen diminishes.
Also examine the adipose tissue slide.
Question 14. In which part of the body will this type of tissue predominate?
Question 15. How many different types of cells can you see?
2. Examine the prepared slide of a regular white fibrous connective tissue. Look at the arrangement
of the fibers. These fibers are predominantly collagen.
Question 16. In which part of the body will this type of tissue predominate?
Question 17. Compared to loose connective tissue, what are the relative amounts of cells,
fibers, and ground substance? How do these relationships adapt each tissue to the particular
job they must perform?
3) Examine the prepared slide of adipose tissue. At first glance, the cells appear empty. That is
because the fat globules that fill the cells have been dissolved during the process of slide
preparation. Can you locate the nucleus of the adipose cells?
Sketch 4. Sketch your observations and label the adipose cells nucleus.
Question 18. What are some of the major functions of the adipose tissue?
Question 19. Why do you think adipose tissue is classified as a connective tissue?
4) Examine the prepared slide of a cartilage. Do you see any fibers? Note the location of the
cartilage cells (called chondrocytes) inside the little cavities called lacunae.
Question 20. How many cells are found in each cavity? If more than one cell is in a cavity,
what do you think this indicates?
Sketch 5. Sketch your observations and label the chondrocytes, lacunae, matrix.
33
Question 21. In which part of the body does cartilage predominate?

The texture of cartilage has been described as "rubbery." You can see that this is true by pressing
your nose from side to side, or you might prefer to bend your ears.
Question 22. What do you think is the advantage, if any, of having some portions of the
skeleton (such as the nose and ears) remain as cartilage throughout the life of an animal, while
other parts are replaced by bone?
Be sure to show your answers and sketches to your TA before you leave today.
34
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
Question 11
Question 12
Question 13
Question 14
Question 15
35
Question 16
Question 17
Question 18
Question 19
Question 20
Question 21
Question 22
Sketch 1
Sketch 2
36
Sketch 3
Sketch 4
Sketch 5
37
Post-lab Homework 3
questions
1. What are the defining characteristics of each of the four major tissue types?
2. What are the differences among the 4 types of epithelial tissues you studied?
3. State where each of the epithelial tissue types are found and what their functions are.
4. What are the structural differences among the types of connective tissues?
5. State where each of the connective tissue types and are found and what their functions are.
6. Describe the tissue types that are affected when you cut your finger with a knife and it bleeds.
Which tissues are involved as the wound heals?
7. Adipose or fat tissues are getting a bad rap these days, but without this tissue, an animal will not
survive. Explain.
8. What areas of the body is cartilage found? What is its functional significance?
38

1. Describe the difference between mechanical and chemical digestion.

2. What molecules are necessary for chemical digestion? Where in your body are sugars broken
down?
3. Name the enzymes that break down proteins.
4. List three enzymes that are found in the small intestines and describe what they do (your answer
should include information about digestion, reagents, color changes, and products).
5. In the exercises that you will perform, how will you be able to test if proteins were digested?
Carbohydrates? Fats?
39
Lab 4
Nutrition and Digestion
Background
The digestive system provides your body with the nutrients, water, and electrolytes required for
life. You consume food, and everything you take in must be digested, or broken down into
nutrients that your body can actually use. Digestion in vertebrates takes place in the alimentary
canal, a digestive tube where food is mechanically and chemically digested. During mechanical
digestion, large pieces of food are ground and crushed into smaller pieces. This action increases the
amount of surface area available for chemical digestion where enzymes break down complex
molecules into smaller molecules. All foods contain at least one of six basic nutrients:
carbohydrates, proteins, lipids, vitamins, minerals, or water. Carbohydrates are broken down to
simple sugars, proteins are broken down to amino acids, and fats are broken down to glycerol and
fatty acids.
When breaking down larger molecules,
digestive enzymes help speed up the reaction.
Chemical digestion takes place in several organs
of your digestive system. Each organ contains
enzymes targeted to breaking down specific
food types. For example, in the mouth, the
salivary glands produce and secrete amylase,
which breaks down starches (carbohydrate) into
disaccharides (units of two sugars). Proteins
start breaking down in our stomach, which has
an acidic pH of around 2-3 due to the
hydrochloric acid. The enzyme in the stomach
that cuts up the bonds between the amino acids
in the protein we eat is called pepsin. Once the
food moves into the small intestine, the
pancreas
releases
sodium
bicarbonate
(NaHCO3), which makes the pH change into a
slightly basic (alkaline) pH of 8. The pancreas
also makes the enzyme called trypsin, which
works well in these conditions and does a similar job to what pepsin did in the stomach. The
pancreas and small intestine produce numerous other enzymes that complete the breakdown of
proteins, lipids, and carbohydrates. The products of chemical digestion are absorbed in the lining of
the small intestine through specialized structures called villi. These nutrients are chemical
substances found in foods that provide energy, serve as building materials, help repair and maintain
cells, and support growth and development of our bodies.
40
Organ
Mouth
Esophagus
Stomach
Small Intestine
Large Intestine
Mechanical Digestion
Crushing
Grinding
Moistening
Moistening
Crushing
Churning
Moistening
Absorption of liquids
Absorption of simple
sugars
Fat emulsification
Food absorption
Chemical Digestion
starch (salivary amylase) maltose
proteins (pepsin) polypeptides
starch (pancreatic amylase) maltose, lactose, sucrose

disaccharide (maltase, lactase, sucrase) monosaccharides
proteins (trypsin, chymotrypsin) peptides
polypeptide (peptidases) amino acids
fats (lipase) fatty acids & glycerol
Water absorption
Exercise 1 Digestion of Proteins

Proteins are long chains of amino acids. Strong bonds called peptide bonds hold these chains
together. Protein digestion begins in the stomach where pepsin breaks proteins into smaller chains
called polypeptide chains. Digestion of proteins continues in the small intestine where the
polypeptide chains are further broken down into smaller peptides by trypsin and chymotrypsin,
and then into amino acids by peptidases. The amino acids are absorbed through the villi that line
your small intestine and are transported to your cells in the blood stream. Your cells put them back
together in different sequences depending on the type of protein needed by the cell.
Materials
-5 clear microtubes
-3 mL pepsin
-5 pH strips
-3 mL biuret solution
-3 mL albumin (water soluble protein)
-3 plastic droppers
-3 mL hydrochloric acid
1) Label your tubes 1-5.
2) Place 4 drops of albumin in tubes 1-4.
2) Add 4 drops of hydrochloric acid to tube 2.
3) Add 4 drops of pepsin to tubes 3 and 4.
41

6) Stir the mixture in each well by inverting the tubes gently several times. Allow the mixture in
each tube to remain undisturbed for 8 minutes.
Question 1. Given this experimental set-up, what do you predict will happen in your tubes?
7) Determine the pH in each of the tubes. Dip a separate pH strip in each tube and compare the
color change to the pH chart provided on the package. Record your results in Table 4.1.
8) Test for the presence of protein by adding 2 drops of biuret reagent to each tube. Carefully
observe any color changes in each of the tubes. A positive test for protein is indicated by a pinkish
color. A negative test will show a blue color that indicates the protein has been digested and broken
down into amino acids. Record your results in Table 4.1.
Question 2. What happened to the protein in tube 2 after HCL was added?
Question 3. What happened to the protein in tube 3 when pepsin was added?
Question 4. What happened to the protein in tube 4 when pepsin and HCL were added?
Question 5. What is the role of pepsin in human digestion? In what parts of the digestive
system does protein digestion take place?
Question 6. What do you call the subunits that make up protein?
Question 7. How could you tell that protein digestion took place?
Question 8. Which of the tubes shows the greatest protein digestion and why?
Exercise 2 Digestion of Carbohydrates

Carbohydrates are large complex molecules that are the body's main source of energy. The digestion
of carbohydrates begins in the mouth. The enzyme amylase, which is produced by your salivary
glands, starts the process of breaking carbohydrates down into smaller molecules. Then your
pancreas releases pancreatic amylase into your small intestine through a small tube and further
breaks the carbohydrates down into simple sugars like glucose and fructose. The sugar molecules
are absorbed through the villi that line your small intestine and are transported to your cells in the
blood stream. Your cells uptake glucose with the help of a hormone called insulin. Your cells then
break glucose down in the mitochondria to produce energy.
42
Materials
-2 clear microtubes
-Amylase solution
-3 plastic droppers
-Starch solution (carbohydrate)
-Potassium iodine solution
1) Label your tubes with the numbers 6 and 7.
2) Place 10 drops of starch solution in tubes 6 and 7.
3) Add 3 drops of amylase solution to tube 7. Allow the mixture to remain undisturbed for 8
minutes.
4. Test for the presence of starch (carbohydrates) in each of the tubes. Add 1 drop of potassium
iodine solution to tubes 6 and 7. In the presence of starch, the mixture will change color to blueblack. If the starch has been digested the solution will remain yellow-brown. Record your results
in Table 4.1.
Question 10. What is the purpose of tube 6?
Question 11. What is the role of amylase in human digestion and where is it produced?
Question 12. Explain what happened to the starch in tube 7 after the addition of amylase.
Question 13. What are the subunits that make up carbohydrates?
Question 14. How could you tell if carbohydrate digestion has taken place?
Question 15. Where does carbohydrate digestion take place?
Exercise 3 Digestion of Fats
Fats are needed by your body to store energy, cushion organs, and help the body absorb vitamins.
The pancreas releases the enzyme lipase into the small intestine where fats are broken down into
fatty acids. The fatty acids are absorbed by the epithelial cells of the small intestine, where they are
then complexed with proteins to form chylomicrons. The chylomicrons diffuse into the lymphatic
vessels.
Materials
-4 clear microtubes
-4 droppers
- DI water
-Corn oil (fat)
-Lipase solution
-4 pH test strips
43
-Liquid soap (fat)

1) Label your tubes with numbers 8-11.
2) Place 7 drops of water and 1 drop of corn oil in tubes 8 - 11. Stir the contents of each tube
thoroughly by inverting the tube several times.
3) Add 1 drop of soap to tube 9
4) Add 4 drops of lipase solution to tube 10.
5) Add 4 drops of lipase solution and 1 drop of soap solution to tube 11.
6) Test the pH of each tube with the pH strip. Compare the color change to the chart. Record the pH
in Table 4.1.
7) Stir the mixture in each tube by inverting the tube several times. Allow the tubes to remain
undisturbed for 8 minutes.
6) Test the pH in each well and record their results and observations in Table 4.1. The lower the
pH the more acidic the solution will be. A high acid content means that the fats are being broken
down into fatty acids.
7) Carefully observe any color changes in each of the tubes. Record your results and observations
in Table 4.1. A clear-yellow solution means that the fat has not been digested. A cloudy solution
means the fats have been broken down.
Question 17. How does the pH of each solution show that fat digestion has occurred?
Question 18. Which tube shows the greatest degree of fat digestion. Explain why.
Question 19. Where does fat digestion take place in the human digestive system?
Question 20. What is the purpose of tube 8?
Question 21. Briefly discuss your results and describe what you have learned today.
Be sure to show your answers to exercises 1-3 and Table 4.1 to your TA before you leave
today.
44
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
Question 11
Question 12
Question 13
45
Question 14
Question 15
Question 16
Question 17
Question 18
Question 19
Question 20
Question 21
46
Table 4.1 Digestion of Macromolecules

Tube #
Contents
pH
Macromolecule test
1
Albumin
Albumin +
HCl
Albumin +
pepsin
Albumin +
pepsin + HCl
HCL
Starch
Starch +
amylase
2
3
7
8
9
10
11
Oil + water
Oil + water +
soap
Oil + water +
lipase
Oil + water +
lipase + soap
47
Observation
Post-lab Homework 4
questions
1. Does the effectiveness of pepsin depend on pH? Explain your answer.

2. Describe the kind of chemical digestion that occurs in the mouth.
3. What are villi and microvilli and why are they important to digestion?
4. Describe the alimentary canal and the type of digestion that occurs in each area. Where does the
majority of digestion occur? Where does the majority of the absorption occur?
5. Describe the relationship between amylase, glucose, insulin, and mitochondria.
6. What is the function of the gall bladder in digestion? The pancreas?
7. What is the primary function of the large intestine?
8. Describe peristalsis.
48

1. Describe the main task of the kidneys. Blood flows into the kidneys via what vessel? Blood
flows out of the kidneys via what vessel?
2. What is the difference between the urethra and ureters?
3. List the three basic mechanisms for separating the various components of the blood.
4. Describe the function of the nephron (use key words: renal corpuscle, loop of Henle, collecting
duct, urea, proximal tubule, distal tubule, Bowmans capsule, filtrate, urine, glomeruls)
5. Why is a urinalysis useful (from a medical perspective). What substances should not be found in
urine (list at least 4)? What would their presence indicate?
49
Lab 5
The Urinary System
Background
Your trillions of cells function, work, and perform tasks that result in the production of a number of
toxic metabolic by-products (carbon dioxide, nitrogen-containing wastes, ammonia, and so on).
These metabolic wastes end up circulating in your blood and must be removed from the body
through a process called excretion. Carbon dioxide is excreted via the lungs, and excess salts and
water are secreted through the skin. It is the function of the urinary system to remove nitrogencontaining wastes from the bloodstream. The urinary system also is responsible for regulating the
chemical make-up, pH, and water balance of the blood. As such, the urinary system is vitally
important to homeostasisand to survival.
In humans, the urinary system is made up of the kidneys, ureters, the urinary bladder, and the
urethra. The paired kidneys are the main organs of the system, filtering and processing blood. The
kidneys are approximately the size of your fist and are located underneath the diaphragm near the
lower back. Nearly a fourth of the total blood flow of the body is delivered to the kidneys each
minute by the large renal arteries. The kidneys allow toxins, metabolic wastes, and excess ions to
leave the body in the urine, while simultaneously retaining necessary substances and returning them
to the blood. After filtering the blood, the blood exits the kidney through the renal vein. The
remaining organs of the system act as transportation channels or storage reservoirs for urine. Urine
leaves the kidneys via the ureters, which carry the urine to the bladder, a muscular bag shaped like
a triangle. The bladder stores urine until the accumulation of urine signals the brain to tell the
bladder that it is ready to release urine. Urine exits the bladder through a tube called the urethra.
Exercise 1 Kidney Filtration
The kidneys have three basic mechanisms for
separating the various components of the blood:
filtration, reabsorption, and secretion. These three
processes occur in the functional units of the
kidney, the nephrons. Each kidney contains
approximately one million nephrons. Each
nephron contains a cluster of blood vessels known
as the glomerulus, surrounded by the hollow
Bowman's capsule. The glomerulus and
Bowman's capsule together are known as the
renal corpuscle. The blood is pushed into the
capillaries of the glomerulus and the force
squeezes liquid (filtrate) out of the blood. Larger
50
units in the blood, such as blood cells and

proteins, remain in the capillaries. From the
glomerulus, the filtrate is passed into a U-shaped
tubule (proximal tubule, loop of Henle, distal
tubule). The filtrate is made-up primarily of urea,
but beneficial molecules also are present and must
be reclaimed by your body. Reabsorption and
secretion occur in this U-shaped tubule. The
tubule is in close proximity to the capillaries (not
shown in the figure) that carry the blood through
the kidneys.
This close proximity allows
diffusion and osmosis to occur. Semipermeable
membranes surrounding the tubule allow selective
passage of beneficial particles (nutrients, NaCl,
H2O, HCO3) back into the blood (reabsorption).
At this point, the blood still contains unwanted molecules. The close proximity between the tubule
and capillaries also allows movement of these unwanted particles (H+ and toxins) from the blood
into the tubule (secretion). The material that has not returned to the blood is called urine and is
shunted to the collecting duct. Urine exits the kidney as collecting ducts from the various nephrons
merge together and ultimately empty into the bladder.
A person can survive with only one kidney, but if both kidneys fail, the person can die from the
buildup of toxic wastes and the lack of regulation of blood pressure, ion concentration, and pH.
Knowing how the nephron functions has allowed us to simulate this function artificially through a
dialysis machine. The persons blood is pumped from an artery and then through tubes that are
made of selectively permeable membranes. The tubes are immersed in a dialyzing solution, similar
to the fluid inside of a nephron. As the blood moves through the tubing, necessary substances
diffuse in and waste substances flow out. The blood is then returned to the patient and the used
dialyzing solution is discarded. In this exercise, starch, glucose, and sodium chloride (salt)
solutions are placed in dialysis tubing, then the dialysis tubing is placed in distilled water
(dialysate). The solution inside the tubing and the dialysate (solution outside of the tubing) will be
tested with Benedict's reagent, iodine reagent, and silver nitrate solution in order to determine which
components have diffused through the differentially permeable membrane and which have not.
Materials required
-250-mL beaker
-12 test tubes
-dialysis tubing (soak at least 30 minutes in advance of use)
-string
-1.0% starch solution
-10% glucose solution
-5% sodium chloride solution
-Benedicts reagent (tests for the presence of glucose)
-Iodine reagent (tests for the presence of starch)
-0.1 M silver nitrate solution (AgNO3) in a dropper bottle (tests for the presence of salt)
-test tube holders
51
-funnel
-50 and 10 mL graduated cylinder
1. Obtain an 8 inch piece of dialysis tubing. Place it in a beaker of distilled water and allow it to
soak for several minutes.
2. Obtain a clean 50 mL graduated cylinder.
3. Remove the dialysis tubing from the beaker of water and tie a tight knot at one end, so the tubing
will hold liquid, and open the other end.
4. Using a 1.0% starch solution, fill the opened tubing a little less than half way. Using a 10%
glucose solution, fill the dialysis tubing to within 1.5 inches of its top. Using a 5% sodium chloride
solution, fill the remainder of the dialysis tubing to within 1 inch of its top.
5. Twist the top of the dialysis tubing closed and tie it with a piece of string.
6. Use distilled water to rinse any spilled solution from the outside of the tubing.
7. Place the tubing, knotted end first, in the 50 mL graduated cylinder. Add distilled water to the
cylinder so that water just covers the top of your tubing.
8. Let the dialysis tubing remain undisturbed for a minimum of twenty minutes.
Question 1: What do you think will happen to the solution inside the dialysis tubing?
Develop a hypothesis and prediction of what will happen and write it down.
While your dialysis tubing is soaking, perform the following tests.
Benedicts Test
1. Using four tubes, add eight drops of distilled water to test tube A, eight drops of glucose solution
to test tube B, eight drops of starch solution to test tube C and eight drops of 5% sodium chloride
solution to test tube D. To each of these test tubes add four drops of Benedict's reagent and mix the
solution.
2. Record the initial color of the solution in each of these test tubes in Table 5.1.
3. Place all four tubes in a hot-water (boiling) bath for three minutes.
4. Record the final color of the solution in each of these test tubes.
5. Keep these solutions in order to compare these results with your experiment results.
Iodine Test
1. Using four test tubes, add eight drops of distilled water to test tube 1, eight drops of glucose
solution to test tube 2, eight drops of starch solution to test tube 3 and eight drops of 5% sodium
52
chloride solution to test tube 4. To each of these test tubes add four drops of iodine reagent and mix
the solution.
2. Record the color of the solution in each of these test tubes in Table 5.1.
Chloride Test
1. Using four test tubes, add eight drops of distilled water to test tube A-1, eight drops of glucose
solution to test tube A-2, eight drops of starch solution to test tube A-3 and eight drops of 5%
sodium chloride solution to test tube A-4. To each of these test tubes add one or two drops of 0.1 M
AgNO3.
2. Record your observations of the solution in each of these test tubes in Table 5.1.
Testing your dialysis experiment for the presence of glucose, starch, and chloride.
After the dialysis tubing has soaked in water for a minimum of twenty minutes.
1. Set up three fresh test tubes and add eight drops of your dialysate to each.
Label one tube DSB to represent dialysate solution with Benedicts. Label the second one DSI
to represent dialysate solution with iodine. Label the third DSA to represent dialysate solution
with AgNO3.
Test your dialysate with Benedicts reagent by adding four drops of Benedict's reagent to the tube
labeled DSB. Mix the solution by gently swirling
2. Set up three additional fresh test tubes. Open you dialysis tubing and add eight drops of liquid
from you dialysis tubing into each test tube.
Label one tube DB to represent dialysis with Benedicts. Label the second one DI to represent
dialysis with iodine. Label the third DA to represent dialysis with AgNO3.
Test the solution inside your dialysis tubing with Benedicts reagent by adding four drops of
Benedict's reagent to the tube labeled DB. Mix the solution by gently swirling.
3. Boil the DSB and DB tubes for three minutes.
4. Compare both of these to your test tubes labeled A, B, C and D.
5. Test your dialysate with iodine reagent, by adding four drops of iodine reagent to the tube labeled
DSI and mixing the solution.
53
6. Test the solution inside your dialysis tubing with iodine reagent, by adding four drops of iodine
reagent to the tube labeled DI and mixing the solution.
7. Compare both of these to your test tubes labeled 1, 2, 3 and 4.
8. Test your dialysate with 0.1 M AgNO3 solution by adding one or two drops of 0.1 M AgNO3 to
the tube labeled DSA.
9. Test the solution inside your dialysis tubing with 0.1 M AgNO3 solution by adding one or two
drops of 0.1 M AgNO3 to the tube labeled DA.
10. Discard the solutions in the proper waste containers and properly dispose of the tubes.
Question 2. What is the purpose of the Benedicts test? Iodine test? 0.1 M AgNO3 test?
Question 3. What molecules diffused through the dialysis tubing?
Question 4. What molecules remained inside the dialysis tubing?
Question 5. Do you think water will move via osmosis out of the dialysis tubing? Why or why
not?
Question 6. Was your hypothesis supported? Why or why not?
Question 7. In what way(s) does this exercise accurately portray the function of the kidneys?
Question 8. In what way(s) does this exercise NOT accurately portray the function of the
kidneys?
Exercise 2 Urinalysis
Approximately 1.0 to 1.8 liters of urine is produced every 24 hours. Urine is the waste product
filtered within the kidney and is made up of many solutes as well as excess water. Urine is a very
helpful tool for doctors when diagnosing different conditions in patients. Urine normally contains
(in order of decreasing concentration) water, urea, sodium, potassium, phosphate ions, sulphate
ions, creatinine, and uric acid. Small quantities of calcium, magnesium, and bicarbonate ions also
are usually present. Abnormally high concentrations of any of these substances may indicate a
problem. To be a valuable diagnostic tool, a urinalysis must be done within 30 minutes of obtaining
a fresh urine sample, or the urine must be refrigerated. The yellow color of urine comes from the
metabolic break-down of hemoglobin, which produces a by-product called urobilin. The pH of
urine ranges from 4.5 to 8.0, but on average it is slightly acidic, with a pH of 6.0. Diet has a
tremendous impact on urine pH, and a diet high in protein increases the acidity, whereas a diet high
in most vegetables and fruits decreases acidity. Specific gravity of urine refers to the relative
weight of a specific volume of urine compared with an equal volume of distilled water. Because 1
ml of distilled water weighs 1 g, water has a specific gravity of 1.000. Urine contains dissolved
solutes and weighs more than water. Urine has a normal specific gravity between 1.001 and 1.030.
At 1.001, the urine is very dilute, which suggests that the person is drinking a lot of water, or
consuming diuretics. This dilute urine could also indicate diabetes or chronic renal failure. High
specific gravity, on the other hand, is normal for people with limited fluid intake. It could also
indicate kidney inflammation. If excessively concentrated, the solutes may precipitate out of
54
solution and form kidney stones. In this experiment, you will perform a urinalysis on three different
samples of urine, testing a variety of different components.
Abnormal Urinary Components

Glucose is not normally present because reabsorption in the tubule is
Glucose
complete. Presence indicates that plasma glucose levels are well above
normal, as in diabetes. Glucose in urine attracts water by osmosis
increasing the volume of urine formed. Diabetics urinate frequently and
are excessively thirsty as a result. The loss of water dehydrates the body
causing a response to conserve water by concentrating urine.
Ketones are by-products of the metabolism of fat. Ketosis and ketonuria
Ketones
are characteristic of starvation, diets low in carbohydrates, various
metabolic disorders, and diabetes.
Protein is not normally present in urine because the glomerulus should
Protein
not allow filtration of plasma protein. In the early stages of glomerular
disease the filtration barrier can become leaky to protein. This generally
increases glomerular filtration and can also lead to increased urine
volume. Renal disease and damage to the glomerular filtration barrier
can result from a bewildering array of conditions. Untreated high blood
pressure is probably the most significant and rapidly increasing cause of
chronic renal failure. Diabetes and a variety of autoimmune syndromes
also result in glomerular damage. Protein in urine might also result from
bacterial infection.
The presence of hemoglobin in the urine is a result of fragmentation of
Hemoglobin
red blood cells. As a result, hemoglobin is liberated into the plasma and
subsequently appears in the kidney filtrate. The presence of hemoglobin
in urine is indicative of hemolytic anemia, transfusion reactions, burns,
poisonous snake bites, or renal disease.
Bilirubin is a breakdown product of hemoglobin. Red blood cells in the
Bilirubin and
body live for about 120 days after which time they are broken down and
Urobilinogen
the hemoglobin they contain is recycled in a complex process that results
in formation of bilirubin. Bilirubin is formed in the liver and excreted
with the bile into the intestine. The presence of bilirubin in the urine
indicates blood levels that are too high, either because excessive
numbers of red blood cells are being broken down (hemolytic anemia infrequently observed) or because the liver is failing to properly
metabolize the hemoglobin (hepatitis, cirrhosis, or bile blockage due to
gall stones). Either situation can result in anemia as valuable blood
protein and iron are lost.
Urobilinogen is produced in the intestine from bilirubin and gives feces
its brown color. Some is reabsorbed into the blood and either excreted
back into the intestine by the liver or excreted by the kidneys of the
urine. Complete absence may indicate renal disease or obstruction of
bile flow in the liver. Increased levels may indicate hepatitis A,
cirrhosis, or biliary disease.
55
Nitrite is a product of the bacterial reduction of nitrate and indicates the

presence of a bacterial infection. Nitrate is produced in the body through
the metabolism of food. Normally there is no nitrite in urine (normal
urine is sterile). Infections in the kidney do not normally produce pain
during urination. Painful urination is typical of lower urinary tract
(urethra and bladder) infections. These infections usually travel up the
urinary tract. The amount of nitrite formed depends on many things
including: the type of bacteria, how long the urine is stored in the
bladder, the presence of nitrate in the diet. Lower urinary tract infections
can travel up into the kidney if left untreated. They may also be sexually
transmitted. Antibiotics are used to treat infections. Bacterial culture
(identification) is important to guide proper antibiotic therapy.
Red and white blood Blood, often referred to as occult blood (meaning mysterious origin)
indicates bleeding somewhere in the urinary system and is an abnormal
cells
finding.
Acidic urine (below 4.5) diabetes, starvation, dehydration, respiratory
pH
acidosis; Alkaline urine (above 8.0) - kidney disease, kidney failure,
urinary tract infection, respiratory alkalosis.
Sediment and crystals are solids found in urine. Common sediments
Sediment and crystals
include dead epithelial cells sloughed from the urinary tract lining, and
crystals of certain minerals that precipitate out of solution. The most
common stone-forming minerals are calcium and oxalate that can lead to
the formation of kidney stones that can obstruct tubules, causing
significant damage and pain.
Materials
-Plastic pipets
-Urine samples (simulated)
-Waste container for pipets
-Waste container for urine
-stopwatches
-Multistix
Nitrite
1) Observe each of the urine samples provided. What do they look like? Record your observations
in Table 5.2.
2) Follow the manufacturers directions for using the Multistix and test each of your urine samples.
Record your results in Table 5.2.
Note: Be prepared to take the readings on several factors at the same time. All readings must be
made during the second minute after immersion, but readings taken after 2 minutes have passed
may be invalid. Pay careful attention to the directions for method and time of immersion.
Question 9. Describe your analysis of urine sample A.
Question 10. Describe your analysis of urine sample B.
Question 11. Describe your analysis of urine sample C.
Be sure to show your tables and answers to exercises 1-2 to your TA before you leave today.
56
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
57
Question 11
Table 5.1 Presence/Absence of Glucose, Starch and Chloride Testing

Test
Initial
Color after
Glucose
Starch
Color
heating
Present (Y/N) Present (Y/N)
Benedicts Test
Distilled water
N/A
(tube A)
Glucose solution
N/A
(tube B)
Starch solution
N/A
(tube C)
Sodium chloride
N/A
solution (tube D)
Chloride
Present (Y/N)
N/A
N/A
N/A
N/A
Dialysate
N/A
N/A
Inside dialysis
tubing
N/A
N/A
Iodine Test
Initial
Color
Color after
iodine
reagent
added
Glucose
Starch
Present (Y/N) Present (Y/N)
Chloride
Present (Y/N)
Distilled water
(tube 1)
Glucose solution
(tube 2)
Starch solution
(tube 3)
Sodium chloride
solution (tube 4)
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
Dialysate
N/A
N/A
N/A
N/A
Inside dialysis
tubing
(continued)
58
Table 5.1 Presence/Absence of Glucose, Starch and Chloride Testing

Test
Initial
Consistency
Glucose
Starch
Chloride
consistency after AgNO3 Present (Y/N) Present (Y/N) Present (Y/N)
added
AgNO3
Distilled water
N/A
N/A
(tube A-1)
Glucose solution
N/A
N/A
(tube A-2)
Starch solution
N/A
N/A
(tube A-3)
Sodium chloride
solution (tube AN/A
N/A
4)
Dialysate
N/A
N/A
Inside dialysis
tubing
N/A
N/A
Table 5.2 Three sample urinalysis

Observations and test Urine Sample A
results
Urine Sample B
Color
Consistency
(transparent or cloudy)
pH
Glucose
Protein
Ketones
Red blood cells
Hemoglobin
Nitrite
Leukocytes
Bilirubin
Urobilinogen
Specific gravity cannot be accurately measured using Multistix
59
Urine Sample C
Post-lab Homework 5
questions
1. What specific part of the kidney does the dialysis tubing represent? What is this parts function?
2. Why is it important that the kidney filters the blood?
3. What type of membrane does the dialysis tubing represent?
4. Give an example of this type of membrane that could be found in your body.
Red
5. If you were a doctor and a patients urinalysis came back with high level of glucose, ketones and
an acidic pH, what diagnosis would you immediately look into?
6. If you were a doctor and a patients urinalysis came back with an alkaline pH and high levels of
albumin, what diagnosis would you immediately look into?
7. Describe how urine is produced and describe the function of nephrons (including the glomerulus,
tubules, and collecting ducts) in the kidneys.
8. How do the kidneys help the body maintain homeostasis?
60

1. A foreign molecule that, when introduced into the body, triggers the production of an antibody
by the immune system is called a(n) _________________.
2. A large Y-shaped protein used by the immune system to identify and neutralize foreign objects is
called a(n) ______________________.
3. Why cant a person with A- blood get a transfusion with AB+ blood?
4. List eight components of the innate immune system.
5. Name and describe the white blood cells that function as part of the acquired immune system.
Be sure to give details of the specific functions for each.
61
Lab 6
The Immune System
Background
All animals have some form of innate
immunity. Innate defenses include external
barriers (the skin, mucous membranes, nostril
hairs, cilia, secretions, etc.) and internal
defenses (macrophages and neutrophils
(phagocytic cells), natural killer cells,
defensive proteins, and the inflammatory
response). In addition, vertebrates have an
acquired immune system, which is highly
specific but requires prior exposure to a
particular pathogen in order to mount an
immune response against it. Once primed, the
acquired immune response provides a strong
defense that is capable of acting against one
specific infectious agent but not another. The
infectious agent that elicits an acquired
immune response is called an antigen. An
antigen is anything that your body recognizes
as foreign and launches an immune response
against. Antigens are recognized by proteins called antibodies, which attach to the antigen to
counteract its negative effects within the body. These antibodies are produced by specialized white
blood cells called B cells, and each antibody is usually specific to only one type of antigen. B cells
are a type of white blood cell known as a lymphocyte. The second type of lymphocyte is the T
cell, for which there are two distinct forms. The cytotoxic T cells attack infected cells, and the
helper T cells activate cytotoxic T cells, B cells and other white blood cells and recruit them to areas
of infection within the body. Following an initial exposure to a particular antigen, memory B cells
and memory T cells are created. These memory cells can survive for years, sometimes even for a
life time. During a second encounter with the same antigen, memory cells can reproduce quickly
and mount a faster and stronger immune response than the first time the immune system responded
to the antigen.
Exercise 1 Immunity
How does your immune system differentiate between self and not self so that it does not attack
the cells of your body? The recognition of cellular markers that are common to our cells make it
possible for the immune system to realize what is foreign. Cellular markers number in the hundreds
and are found on the surface of our cells. For example, red blood cells have markers called
agglutinogens which can act as antigens. Each of us carries antibodies within the plasma of our
blood that attack agglutinogens that are not found in our own blood cells. The antibodies are called
62
agglutinins. When foreign agglutinogens are found in our blood, agglutinins connect to the red
blood cells and mark them for destruction by the immune system. Connection of the agglutinins to
the agglutinogens is called agglutination and it forms clumps that can be visualized. The reaction is
highly specific because it depends on the specific antibody and antigen pair.
Agglutination reactions have lots of applications in medicine, such as blood typing cells for
transfusion, identifying bacterial cultures, and in detecting the presence and relative amount of a
particular antibody in a persons blood. Agglutination has been used to determine if a patient is
suffering from, or has suffered from, a bacterial infection. For example, if a person is suspected of
having typhoid fever, the persons blood serum is mixed with a culture of Salmonella typhi. If
agglutination occurs, visible as clumps of bacteria, the patient either had or has an S. typhi infection.
In this lab exercise, you will use a basic blood typing kit to determine your blood type.
There are two unique agglutinogens (antigens)
on human red blood cells, they are called either
A or B antigens. These antigens are inherited
youve received one from each parent - so you
can have up to two different antigens in your
blood. Some people have a mutation that
prevents them from producing a normal,
functioning agglutinogen. These non-functional
types are called O. Each person can carry
either anti-A and/or anti-B antibodies in his or
her blood dependent on his or her blood type
(some people carry both, some carry
neither). When a person receives unmatched
blood, these antibodies will agglutinate blood
cells, causing blood clotting and possibly death.
The RhD antigen is another important aspect in determining blood type. The term "positive" or
"negative" refers to either the presence or absence of the RhD antigen. Anti-RhD antibody is not
usually a naturally occurring antibody as the Anti-A and Anti-B antibodies are.
Blood Types
Genotype
Antigens present
Antibodies present
Type A
AA or AO
A
Anti-B
Type B
BB or BO
B
Anti-A
Type O
OO
None
Anti-A and Anti-B
Type AB
AB
Both A and B
None
Question 1. Given the table above, consider each of the blood types and determine if
agglutination will occur if they receive A blood type. Discuss why/why not.
agglutination will occur if they receive B blood type. Discuss why/why not.
agglutination will occur if they receive O blood type. Discuss why/why not.
63
agglutination will occur in the presence of AB blood type. Discuss why/why not.
Question 5. Does a person who is RhD(-) produce RhD antibodies? What would happen if
this person received Rh(+) blood?
Question 6. Does a person who is RhD(+) produce RhD antibodies? What would happen if
this person received Rh(-) blood?
SAFETY NOTE: This lab uses your own blood for ABO blood typing. Care must be taken to
prevent self-infection from environmental contaminants and the transmission of blood-borne
infections, such as viruses causing hepatitis and AIDS. Typing of your own blood maybe done
safely if you rigidly follow the safety instructions, typing procedures, and your TAs instructions.
You will be the only person who will be in contact with your blood. After obtaining your drops of
blood, put on gloves.
If you are not a blood donor, your job is to make sure that no one but the blood donor comes into
contact with his/her blood and that all items used are properly disposed of IMMEDIATELY and
NOT set on the lab benches. Dispose all waste in the biohazard container.
SAFETY PRECAUTIONS
-Avoid contact with blood of another student.
-Before piercing your finger, wash your hands with soap and water and disinfect your fingertip with
an alcohol wipe.
-Do not remove the sterile lancet from its container until you are ready to use it. Do not touch its tip
or lay it down.
-Use a lancet once and immediately place it in the biohazard sharps container provided. Never place
a used lancet on the tabletop or in the wastebasket.
-Place all other materials in contact with blood, such as alcohol wipes, spatulas, and tissue in a
biohazard bag immediately after use.
-When finished, wash the tabletop around the typing box with a disinfectant. Wash your hands with
soap.
-Follow your TAs directions at all times.
Testing for antigens in your blood is accomplished by adding antisera containing specific antibodies
that will combine with these antigens. You will be given the following materials:
Materials
- Slide Guide
- One blue and one yellow spatula
- One sterile disposable blood lancet
- One disposable alcohol pad
- Anti-A sera
- Anti-B sera
64
1. Place the slide guide on a flat area in front of yourself.

2. Addition of ANTI-SERA
Place one small drop of anti-A sera (blue) in the proper circle on the slide guide, and one
small drop of anti-B sera (yellow) in the other circle of the slide guide. Place these drops to
the far right side of the circles in such a way that give you plenty of room to deposit a drop
of blood to the slide guide without making contact with the anti-sera. (As shown in the
figure below).
3. Prepare to draw blood

Swing the non-writing hand vigorously several times. Swinging the arm several times just
prior to puncturing increases the amount of blood at the fingertips.
Scrub the middle finger of the non-writing hand with the alcohol pad. Allow to air dry. DO
NOT CONTAMINATE THIS FINGER.
Press on the bottom of the fingertip with the thumb of the same hand (to help hold blood in
the fingertip). Using the sterile lancet, puncture the tip of the finger. DO NOT LANCET
ANYONE OTHER THAN YOURSELF. When finished with your lances, immediately
place them in the biohazard container.
Wipe away the first bit of blood with the alcohol pad (this is primarily serum)
Place a drop of blood in each circle on the slide guide. Blood can be squeezed out with a
repeated rolling, pressing motion of the thumb toward the fingertip. DO NOT MAKE
CONTACT WITH THE ANTI-SERA.
Place the alcohol pad over the punctured finger and hold in place with thumb. At this point,
put on gloves. DO NOT TOUCH ANYONE ELSES BLOOD.
4. Carefully (but quickly) mix the blood and the anti-A sera with the blue spatula, and the blood
and anti-B sera with the yellow spatula. DO NOT CROSS CONTAMINATE by using the same
spatula for both. Properly dispose of the spatulas as soon as you are finished with them.
5. Observations
As you mix, watch for results. Examine for agglutination. The agglutination will appear as
the grainy clumps of red blood cells suspended in a clear solution. Do not observe for more
than three minutes. Observations may be confirmed with a small hand lens.
6. Interpretation
Observe for agglutination on both sides of the slide guide
If agglutination (clumping) occurs in both mixtures, the red cells contain both agglutinogens
(antigens) and the blood is obviously type AB. Conversely, if neither mixtures agglutinate,
both agglutiongens are absent and the blood type is O. Type A and type B will agglutinate
only on the side treated with Anti-A serum and Anti-B serum, respectively.
65
Question 7: What is the blood type of the person in your group who was typed?
Question 8: Assume Student A used the agglutination test to determine blood type and got the
following result:
Anti-serum A
Agglutination -
B
+
Rh
+
a) What is Student A's blood type?

b) If Student A receives type AB blood for blood transfusion, what is going to happen and
why?
Exercise 2: Disease Transmission

We are surrounded by thousands of things that might make us sick, including bacteria, viruses, and
parasites that we may get from other people. When an illness can be passed from person to person,
it is considered infectious. Diseases are transferred from person to person in a variety of ways
through the air in droplets when one sneezes, by direct bodily contact, or by touching a surface such
as a doorknob that was recently handled by a sick person.
Through millions of years, humans have evolved ways to combat pathogens, such as bacteria,
viruses, or parasites that cause diseases. The study of the transmission of disease in populations is
called epidemiology and it forms the basis for public health strategies and interventions. An
epidemiologist determines the origin of an outbreak of infectious disease, for example, so that the
source (such as a contaminated well or food) is identified and the public can be educated about how
to prevent infection.
Epidemiology is only one branch of the study of disease. Specifically, it is the study of the spread of
disease. Diseases such as the flu, the common cold, or intestinal infections are often transferred
from person to person by bodily contact, or by touching an object the person has touched. This
exercise is designed to demonstrate how an epidemiologist might track the spread of a
communicable disease back to the original source. We will use glucose, which does not cause
disease, as a model for examining how microbes may be passed from person to person.
Procedure:
1. Each person will receive a small test tube that is identified with a number and filled half way with
a liquid. This liquid will represent bodily fluids. All but one of the dishes will contain water. The
exception will be one containing a solution of glucose. This solution represents the "disease" we
will be tracking. Write down your I.D. number in Table 6.1.
2. You will exchange liquids with another student in the room. Pour all of your liquid into the other
persons test tube do this a few times to thoroughly mix the liquids. This will simulate physical
66
contact and will represent the pathway of infection for this disease. Pour half back into your own
tube. Write the ID number and name of the person with whom you exchanged liquid.
3. Wait until everybody in the room has exchanged liquids once before going on the next round.
Repeat your exchange of liquid with a new student. Keep track of your contacts' I.D. numbers and
the order in which you came in contact with them.
4. Wait until everybody in the room has exchanged liquids this second time before going on the
third and final round (only do this step if there are greater than 8 students in the lab). Repeat your
exchange of liquid with a new student. Keep track of your contacts' I.D. numbers and the order in
which you came in contact with them.
Once everyone has finished bring your tube to the front of the room for diagnosis. We will use
Benedict's reagent to test for the presence of the "disease" (glucose) by placing 6 drops in your tube
and then placing all the tubes in a boiling bath for three minutes . Contact with the "disease" will be
indicated by a color change - A greenish color indicates about 0.5% concentration; yellow
precipitate indicates 1% concentration; orange indicates 1.5% and red indicates 2% or higher
concentration. What happened to the fluid in your tube?
5. If you were diagnosed with the disease, report the I.D. numbers of those you came into contact
with. We will use this information to determine the initial carrier of the disease. Data from the class
will be provided by your TA.
Question 9: How did you determine who was the initial carrier of the "disease"?
Question 10: Was it possible to know precisely who the initial carrier was? Why or why not?
Question 11. Who were the possible initial carriers?
Exercise 3: Lymphatic System
The lymphatic system consists of a network of lymphatic vessels, lymphatic tissue, lymph nodes,
and a number of lymphoid organs, such as the tonsils, thymus and spleen. Overall, the function of
the lymphatic system is to: (1) remove excess fluid, waste, debris, dead blood cells, pathogens,
cancer cells, and toxins from cells and the tissue spaces between them, (2) aid the immune system in
destroying pathogens and filtering waste so that the lymph can be safely returned to the circulatory
system, and (3) work with the circulatory system to deliver nutrients, oxygen, and hormones from
the blood to the cells that make up the tissues of the body.
Lymph originates as plasma, which is the fluid portion of blood. The arterial blood that flows out of
the heart slows as it moves through a capillary bed. This slowing allows some plasma to leave the
arterioles and flow into the tissues where it becomes tissue fluid. Also known as extracellular
fluid, this fluid delivers nutrients, oxygen, and hormones to the cells. However, this fluid must
eventually return to the blood if the vascular system is to operate properly (if it doesnt, fluid
accumulates in the tissues, producing a condition called edema). As this fluid leaves the cells, it
takes with it cellular waste products and protein cells. It is the tiny lymphatic capillaries that pick
67
up this lymph fluid and carry it through successively larger vessels lymphatic collecting vessels
to lymphatic trunks until the lymph finally returns to the blood vascular system through one of
the two large ducts in the thoracic region. The right lymphatic duct drains lymph from the right
side of the head and neck, the right arm, and the upper right quadrant of the body. This duct empties
the lymph into the right subclavian vein. The thoracic duct drains lymph from the rest of the
body, including both legs. The thoracic duct empties the lymph into the left subclavian vein.
The bloodstream is pumped by the heart. It circulates throughout the body and is cleansed by being
filtered by the kidneys. The lymphatic
system does not have a pump to aid in
its flow, instead this system is designed
so that lymph only flows upward
through the body traveling from the
extremities (feet and hands) and upward
through the body toward the neck. As it
travels through the body, lymph passes
through bean-shaped lymph nodes
where it is filtered.
There are between 600-700 lymph
nodes present in the average human
body. It is the role of these nodes to
filter the lymph before it can be returned
to the circulatory system. Although
these nodes can increase or decrease in
size throughout life, any node that has
been damaged or destroyed, does not
regenerate.
Within
the
lymph
nodes
are
macrophages, phagocytes that destroy
bacteria, cancer cells, and other foreign
matter in the lymphatic fluid, thus
rendering many harmful substances or
cells harmless before the lymph enters
the bloodstream. Although we are not
usually aware of the filtering and the
protective nature of the lymph nodes,
most of us have experienced swollen
glands during an active infection. This
swelling is a manifestation of the
trapping function of the nodes, and due
to the increased numbers of white blood
cells in the body.
68
Lymphoid organs, including the thymus, lymph nodes, spleen, tonsils, appendix, and bone marrow,
also defend the body against invading pathogens. The thymus and bone marrow are considered
primary lymphoid organs because they give rise to lymphocytes (B cells in the bone marrow, T
cells in the thymus). The other lymphoid organs are secondary lymphoid organs and they maintain
and activate nave lymphocytes.
1. Obtain a compound microscope and prepare slides of a lymph node and spleen. As you examine
the lymph node slide, notice the following:
A. The node is enclosed within a fibrous
capsule, from which connective tissue
(trabeculae) extend inward to divide to
node into several distinct parts.
B. In the outer region of the node, the
cortex, some of the cells are arranged in
globular masses, referred to as germinal
centers. The germinal centers contain
rapidly dividing B cells. The rest of the
cortical cells are primarily T cells that
circulate non-stop, moving from the blood
into the node and then exiting from the
node in the lymphatic stream.
C. In the internal portion of the gland, the
medulla, the cells are arranged in a cordlike
fashion. Most of the cells in the medulla are macrophages. Macrophages are important not only for
their phagocytic function, but also because they play an essential role in presenting the antigens to T
cells.
D. Lymph enters the node through a number of afferent vessels, circulates through lymph sinuses
within the node, and leaves the efferent vessels at the hilus. Since each node has fewer efferent
than afferent vessels, the lymph flow slows down dramatically within the node. This allows time
for the generation of an immune response and for the macrophages to engulf debris from the lymph
before it reenters the blood vascular system.
Sketch 1. In the space provided, sketch the structure of the lymph node as you see it in the
slide. Label all parts and draw arrows that indicate the flow of lymph into and out of the
lymph node.
Question 12. In what way(s) does the slide of the lymph node you are looking at differ from
the drawing above?
69
2. As you investigate the slide of the spleen,

look for the areas of lymphocytes suspended in
reticular fibers, the white pulp, clustered around
central arteries. The remaining tissue in the
spleen is the red pulp, which is composed of
venous sinuses and areas of reticular tissue and
macrophages called the splenic cords. The
white pulp, composed primarily of lymphocytes,
is responsible for the immune functions of the
spleen. Macrophages remove worn-out red
blood cells, debris, bacteria, viruses, and toxins
from blood flowing through the sinuses of the
red pulp.
Sketch 2. In the space provided, sketch the structure of the spleen as you see it in the slide.
Label all parts, including the red pulp and the white pulp. Show your TA the white pulp on
your slide for TA initials.
Question 13. Consult the unknown lymphoid tissues and properly identify them.
70
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8a
Question 8b
Question 9
71
Question 10
Question 11
Question 12
TA initials
Question 13
Table 6.1 Microorganisms and Disease Lab Data Sheet

ID Numbers and Names Your Name:
of those you contact:
Your ID#
1
2
3
Sketch 1.
Sketch 2.
72
Post-lab Homework 6
questions
1) Describe the difference between a B cell and a T cell. What are the different types of T cells and
what do they do?
2) What is the difference between innate and acquired immunity?
3) Describe a memory cell.
4) Describe the human ABO blood type and describe how it is important in terms of immunity.
5) Describe the human RhD factor.
6) Describe the results of your blood typing analysis and discuss the meaning of your results.
7) Which ABO blood type may receive blood from the other three in emergencies (the universal
recipient). Why is this the case? Which ABO blood type may donate blood to the other three types
in emergencies (the universal donor). Why is this the case?
8) If a person had antibodies to the West Nile virus in his/her plasma, what would you conclude?
73

1. What is the primary cell type of nervous tissue? Describe the function of each of the following.
a. axon
b. dendrite
c. cell body
2. What is a myelin sheath and what is its function?
3. Define hypothyroidism and hyperthyroidism.
4. Which gland does the hypothalamus directly communicate with?
5. Using the background data on the endocrine system, fill in Table 7.1 before class (but dont hand
it in to your TA yet because you will be using it in class today).
74
Table 7.1 Comparison of Hormonal Effects on Different Organs

Complete before coming to class. Determine what the impact of excess of each of these hormones
would have on these different body parts. Use your lab manual to fill in the rest of the table (where
there are blanks). You will have to show this to your TA when you come into class.
Testosterone
LH
TRH
TSH
ACTH Cortisol
Intact
Castrate
Intact
Castrate
Pituitary
Thyroid
Adrenal
Thymus
Testes
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NA
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
+
+
+
NC
Prostate
Seminal
NC
NC
NC
NC
+
+
+
NC
vesicles
Body
+
NC
weight
Fill in with a (+) to denote an increase in size. Use (-) to denote a decrease in size. Place the letters
NC in the box where no change occurs. TRH, thyroid-releasing hormone; TSH, thyroid-stimulating
hormone; ACTH, adrenocorticotropin hormone; LH, luteinizing hormone.
75
Lab 7
The Endocrine and Nervous Systems
Background
Your nervous system is fast and direct it uses neurotransmitters and neurons to convey
information to and from the brain. In contrast, your endocrine system is much slower it uses
hormones, which are chemical messengers produced by specific tissues in the body, to transmit
information. These hormones travel through the bloodstream to exert their effects on distant target
organs. Hormones are a slower method of communication, but their effects last longer. Today, we
are going to consider the nervous and endocrine systems separately, but please realize that these two
systems are highly interconnected.
Exercise 1 The Nervous System
(adapted from Devlin-Kelly, Bakersfield College)
The nervous system consists of the brain, spinal cord, all nerves, nerve plexuses, nerve roots, and
ganglia. The divisions of the nervous system include the (1) Central Nervous System (CNS) which
consists of the spinal cord and brain, and the (2) Peripheral Nervous System (PNS) which consists
of all nerves, plexuses, roots, and ganglia in the somatic division (skin and skeletal muscle) and the
autonomic division (smooth muscle, cardiac muscle and glands).
The primary cell type of the nervous system is the neuron. Neurons are very unique cells, both in
their appearance and their way of working. A neuron consists of a nerve cell body, dendrites,
which carry impulses toward the nerve cell body, and axons, which carry impulses away from the
cell body. An axon can extend several meters long. Neurons receive signals from either the external
or internal environment, and transmit them in the form of electrical impulses to other neurons,
muscles or glands. There are three types of neurons: sensory neurons, motor neurons, and
interneurons. Sensory neurons lie mostly outside the brain and spinal cord, and receive information
and relay it to the central nervous system. Motor neurons lie mostly outside the brain and spinal
cord, and relay signals from the central nervous system to effector organs (muscles or glands).
Interneurons lie entirely within the central nervous system, and transmit signals within the central
nervous system.
Protective cells (glial cells) surround many nerve processes. Such protective cells, which make up
about 90% of nervous tissue cells, are called neuroglia when they occur in the central nervous
system (brain and spinal cord), and Schwann cells when they occur in the peripheral nervous
system. These cells contain a fatty substance, myelin, which forms many layers around the fibers,
and functions to insulate and to increase the rate of transmission of impulses along the fibers. The
myelin sheath is interrupted at regular intervals by openings called the Nodes of Ranvier. Impulses
traveling along myelinated fibers jump from node to node, resulting in a tremendous increase in
velocity of the impulse.
76
Materials
-Neuron cell models
-Neuron axon kits (1/person) consisting of:
Zip-loc bag
1 round hard candy
pencil
Scotch tape
-Compound microscopes
-Slides:
Motor Neuron
Cross section and longitudinal section of myelinated axon
Cross section of spinal cord
-Spinal Cord Models
1) Label the neuron diagram in Figure 7.1 using your text, lab atlas (if available) or lecture notes for
reference.
Be sure to label the following structures:
Neuron cell body
Nucleus
Axon
Dendrites
Schwann cell
Schwann cell nucleus
Myelinated sheath
Nodes of Ranvier
2) Use a pencil to represent the axon of a neuron. Put a round hard candy into a Ziploc bag and zip
it leaving a small amount of air inside. The Ziploc bag is a Schwann cell.
Question 1. What does the candy represent?
3) Now place the bottom of the bag (Schwann cell) along the length of the pencil (axon) and tape it
there. Begin to wind the bag around the pencil until it is completely wound up. Scotch tape the top
of the bag in place to hold it on the pencil. You now have built a myelinated axon. Nerve impulses
can travel down the length of this axon. Do you see how the Schwann cell is mostly cell membrane?
4) Now put your axon down in the middle of the lab table. Line up all the axons at your table end to
end (like beads on a string). You now have a longer axon, made up of several pencils and Schwann
cells. Find the nodes of Ranvier.
Question 2. What is the advantage of being a myelinated axon versus a non-myelinated one?
Question 3. How does a myelinated axon transmit information more rapidly than a nonmyelinated one?
77
5) Observe the neuron models in the lab (if available). Have your lab partner quiz you on the parts
of the neuron until you can identify all the parts above without looking at your books.
6) Using your compound microscope, observe the following slides and sketch them in the space
provided.
Sketch 1. View the motor neuron and sketch what you see. Be sure to label structures.
Sketch 2. View the cross section of the human nerve and sketch what you see. Be sure to label
structures.
Question 4. View the longitudinal section of the human nerve. Do you observe any differences
between this and the model in the room? Why are there differences?
Anatomy of the Spinal Cord.
When you slice the spinal cord in a transverse or cross section you see a structure which resembles
a butterfly. This pattern is visible because the spinal cord is made of two different kinds of neurons
which make up the grey matter and white matter. The grey matter consists of non-myelinated
neurons. The white matter consists of myelinated neurons.
7) Observe a slide of a spinal cord cross section on low power. First find the central canal. This
will be an opening in approximately the center of the entire spinal cord. Find the butterfly-shaped
structure which surrounds the canal. This is the grey matter. Within this region are numerous nerve
cells or neurons. The nerve cells are large and irregular in shape because of the various processes
projecting out of the cells body. The remainder of the cord is the white matter.
8) The fanning out portions of the grey matter (the wings of the butterfly) are the horns. Then
determine the anterior and posterior of the cord. In the anterior, are the ventral horns. This is the
location of the greatest density of neuron cell bodies. These are multipolar neurons and they
78
contain the cell bodies of the efferent motor neurons which extend out to contact the skeletal
muscle cells. Look for the following and be prepared to show each to your TA:
a. Ventral horn
b. Dorsal horn
c. Gray matter
d. White matter
e. Central canal
f. Motor neuron in gray matter
Question 5. Your TA will ask you to show him/her one of the above for TA initials.
9) Observe the clump of tissue which is located dorsally outside the spinal cord. This is the dorsal
root ganglion. The cell bodies located here bring sensory information from the body to the central
nervous system.
Exercise 2 The Endocrine System
(adapted from Odenweller, Hsu, Sipe, Layshock, Varyani, Rosian and DiCarlo 1997)
The command center for the endocrine system is the hypothalamus, a small, penny-sized portion of
the brain. The hypothalamus acts as an endocrine organ that secretes oxytocin and anti-diuretic
hormone (ADH, also known as vasopressin). These hormones travel down the pituitary stalk to the
posterior pituitary gland where they are released directly into the bloodstream. In addition, the
hypothalamus also regulates anterior pituitary gland function through the secretion of releasing
hormones: thyroid-releasing hormone (TRH), corticotropic-releasing hormone (CRH), and
gonadotropic-releasing hormone (GnRH). These releasing hormones stimulate the synthesis and
secretion of anterior pituitary hormones, which include thyroid-stimulating hormone (TSH),
luteinizing hormone (LH), follicle stimulating hormone (FSH), growth hormone (GH),
adrenocorticotropic hormone (ACTH), and prolactin.
Each of these hormones is released into the bloodstream to affect specific target organs. For
example, the hypothalamus secretes TRH, which travels to the pituitary gland to release TSH; TSH
travels to the thyroid gland (the target organ) and stimulates the release of thyroid hormone. It is
important to note that the hypothalamic releasing hormones are only required for the synthesis and
release of the anterior pituitary hormones. The posterior pituitary hormones are synthesized by the
hypothalamus and travel down neurons to be released from the posterior pituitary gland. Because
the anterior pituitary gland secretes multiple hormones, it is frequently referred to as the master
gland. For this experiment, we will focus on the hypothalamus only as a regulator of the anterior
pituitary gland. Figure 7.2 shows the relationship between the hypothalamus and the pituitary gland.
In the endocrine system, negative feedback is used to inhibit further hormone secretion. When a
sufficient amount of hormone has been released, it communicates or feeds back to suppress the
releasing organ. In other words, the gland has released enough hormone to fulfill its function; this is
sensed by the body, and production of the hormone ceases. Negative feedback not only inhibits the
releasing organ, but can also inhibit the pituitary gland and/or hypothalamus. By using a negative
feedback system, the body produces only the amount of hormone it needs without wasting its
resources. Conversely, in positive feedback, the end product further stimulates the releasing organ.
This form of feedback is less common.
79
Figure 7.2. Secretion of hypothalamic hormones. Hypothalamic releasing hormones travel down the hypothalamic
hypophysial portal system to the anterior pituitary gland, where they stimulate the synthesis and release of anterior
pituitary hormones [adrenocorticotropin hormone (ACTH), thyroid-stimulating hormone (TSH), follicle-stimulating
hormone (FSH), luteinizing hormone (LH), growth hormone (GH), and prolactin]. In contrast, the posterior pituitary
gland does not require releasing hormones, because the hypothalamus synthesizes and secretes both anti-diuretic
hormone (ADH) and oxytocin.
The pathways of three hormones are examined in this experiment: thyroid hormone, cortisol, and
testosterone. The hormonal pathways are similar in all three cases. It is important to realize that the
hypothalamus secretes a releasing hormone to regulate each of the hormones secreted from the
anterior pituitary gland. In this way, the hypothalamus is like a command center. If the
hypothalamus is not stimulated, the hypothalamic releasing hormones (TRH, CRH, and GnRH) will
not stimulate the anterior pituitary gland to secrete its hormones.
The hypothalamus releases TRH, which travels to the anterior pituitary gland via the bloodstream to
stimulate production of TSH. TSH travels to the thyroid gland (located by the trachea) to stimulate
the production and release of thyroid hormone. Thyroid hormone influences the growth rate of
many body tissues and is necessary for proper central nervous system development. Its main
function is to increase a persons basal metabolic rate (BMR) and to increase heat production. An
excess of thyroid hormone can negatively feed back to inhibit further thyroid hormone release from
the thyroid gland, TSH secretion from the anterior pituitary gland, and/or TRH release from the
hypothalamus.
Similarly, ACTH is released from the anterior pituitary gland in response to CRH secreted from the
hypothalamus. ACTH stimulates the adrenal glands (located on top of the kidneys) to secrete
80
cortisol, which promotes the breakdown of proteins and fats and helps the body adapt to stress.
Cortisol functions to provide the body with fuel by breaking down (catabolism) the materials of the
body. Under normal conditions, excess cortisol in the bloodstream will negatively feed back to the
hypothalamus (to inhibit CRH release), anterior pituitary gland (to inhibit ACTH secretion), and/or
to the adrenal gland (to inhibit further cortisol release). The release of CRH is regulated by negative
feedback, circadian rhythms, and stress. Cortisol can also act as an immunosuppressive and antiinflammatory agent. If cortisol is administered in large doses, its immunosuppressive properties will
cause the organs of the immune system to shrink. In this experiment, the thymus gland will
represent the organs of the immune system.
Question 6. Describe the effects of thyroid hormone.
Question 7. Describe the effects of cortisol.
LH is released from the anterior pituitary gland in response to GnRH secreted from the
hypothalamus. LH is seen in both males and females but has different functions. In the male, LH
travels to the Leydig cells that are located in the connective tissue between the seminiferous tubules
of the testes. The Leydig cells release testosterone, which is responsible for the male sex drive and
secondary sex characteristics, such as increased body hair and a deeper voice. An excess of
testosterone can cause an increase in muscle mass. Negative effects of testosterone are male pattern
baldness and increased secretion of the sebaceous glands, which can lead to acne. In the female, LH
causes the developing egg in the ovary to secrete estrogen. Estrogen participates in either a positive
or negative feedback loop, depending on the stage of the menstrual cycle. In the preovulatory and
postovulatory phases, estrogen regulates the release of LH through negative feedback. However,
there is a large rise in levels of LH just before ovulation (release of the egg from the ovary) due to a
positive feedback mechanism. During this interval, the secretion of estrogen from the follicle further
stimulates the release of LH from the anterior pituitary gland. The increased levels of LH are
essential for ovulation to occur. Estrogen causes the development of female secondary sex
characteristics and sustains the female reproductive tract. A woman who lacks ovaries will not
produce estrogen. However, the pituitary gland will secrete excess LH because the feedback
inhibition no longer exists. Excess levels of estrogen cause early sexual development in the female
as do high levels of testosterone in males.
Question 8. Describe the role of LH in both males and females.
To simplify the relationship between the reproductive and endocrine systems, we will concentrate
only on the male system. The female reproductive system is more difficult to study than the male
reproductive system because it is continuously cycling. The pathways of all three hormones can be
understood by looking at a visual representation in Figure 7.3 (Figure 7.3 also demonstrates the
pathways of the hormones that will be used throughout the experiment, thus serving as an aid in the
analysis of laboratory data). The glands and tissues of our body enlarge (increase in size) if they are
continuously activated; this is called hypertrophy. For example, a person who lifts weights will
continually stimulate the activated muscles, resulting in hypertrophy. This can be easily observed
when comparing a bodybuilder to an average person; the bodybuilders muscles appear larger in
comparison. In contrast, if a gland or tissue is continuously inhibited it will shrink in size or
atrophy. For example, if a cast is placed on a persons arm for six weeks and then removed, a
81
drastic reduction in muscle mass can be seen. The cast prevented any movement (stimulation) of the
limb, allowing atrophy to occur.
Figure 7.3. Negative feedback control (hormone pathways). GnRH, gonadotropin-releasing hormone.
Question 9. Explain the positive feedback loop observed in LH regulation.

Question 10. Describe the difference between hypertrophy and atrophy.
There are many diseases that may result from a deficiency or excess of hormones. These hormonal
imbalances may lead to changes in organ or gland size (hypertrophy or atrophy). Hyperthyroidism
is the excessive production of thyroid hormone. The most common cause of hyperthyroidism is
Graves disease; the symptoms include increased BMR, a constant feeling of warmth, nervousness,
and an enlarged thyroid gland (known as goiter). In contrast, hypothyroidism is the result of
decreased levels of thyroid hormone. A patient with hypothyroidism will present symptoms of low
BMR, a decreased appetite, abnormal central nervous system development, and an intolerance to
cold. Cushings syndrome is the result of excess secretion of cortisol (hypercortisolism). The
symptoms of Cushings syndrome include personality changes, hypertension (high blood pressure),
osteoporosis (weakening of bones due to loss of calcium), and weight loss. If an excess level of
cortisol remains in the body, protein degradation will occur leading to a wasting effect.
Hyposecretion (decreased secretion) of cortisol is characterized by symptoms such as defective
metabolism, mental confusion, and a decreased ability to adapt to stress. Decreased amounts of
testosterone in the body primarily affect the sexual organs. If testosterone levels are low, males will
not develop normally and will have sperm counts too low to fertilize an egg. The condition of
excess levels of testosterone is rare but causes premature sexual development.
Question 11. Consider the differences between hyperthyroidism and hypothyroidism. What
are some characteristics of each?
82
Question 12. What are the effects of decreasing testosterone?

The data for this laboratory were compiled from seven sets of male laboratory rats, two rats per set;
one set was the control group and the remaining six were experimental groups. The rats were all
male to simplify the study of the relationship between the reproductive and endocrine systems. In
each set of rats there was an intact rat and a castrate rat. The castration involved removal of
the testes to eliminate testosterone production. The two rats (normal and castrate) of each group
were treated alike in all other ways (food, water, etc.). All rats, except for those in the control group
were injected with a hormone on a daily basis for 2 wk. Autopsies were performed on the animals at
that time.
Unfortunately, the group of students performing this exercise were very disorganized and rushed
through the work, making errors in labeling the bottles of hormone. The students obtained the
following results for organ weights after the autopsies were performed. In this short period of time,
the students noted amazing changes in the size of certain organs when they compared the
experimental group of rats with the control group. Before coming to class, you should have filled
out Table 7.1. Now, using Table 7.1 and the autopsy data in Figure 7.5, match the unknown rat
groups with their respective hormones. The bottles on the refrigerator shelf were ACTH, cortisol,
LH, TSH, TRH, and testosterone.
To help in determining the identity of the unknown hormones, you should look for changes between
the control values and the values of the unknown hormone (both the intact and castrate animal). The
changes between the control rats and the rats that were treated with the unknown hormone should
be >20% if they are to be considered significantly different. If the change is <20%, it is attributed to
experimental or biological error. Experimental errors may include small errors in calibration
procedures, measurements, or instrumentation. Any variability that occurs because of the
differences between animals is considered biological error.
Figure 7.4. Graphic representation of organs studied in the autopsy.
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Figure 7.5 Autopsy results from control rats and six experimental rats
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85
86
87
Determine the identity of hormones 1-6 using the data from the autopsy and Table 7.1.
Question 13. What was hormone 1? Explain your answer.
Be sure to show your Figures, Table, drawings and answers to your TA before you leave
today.
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Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
TA initials
Question 6
Question 7
Question 8
Question 9
Question 10
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Question 11
Question 12
Question 13
Question 14
Question 15
Question 16
Question 17
Question 18
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Figure 7.1 Unlabeled Neuron
Sketch 1
Sketch 2
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Post-lab Homework 7
questions
1) What is the function of neurons?
2) Describe the structure and function of a Schwann cell.
3) Explain why part of the spinal cord looks grey and part looks white.
4) What is a ganglion? What is a neuromuscular junction?
5) Describe the hypothalamus and the role it plays in the autonomic nervous system and the
endocrine system.
6) List the hormones released by the anterior pituitary gland.
7) Describe ACTH, cortisol, LH, TSH, TRH, and testosterone by describing where they are
produced in the body and their physiological functions.
8) Why is the nervous system considered fast and the endocrine system considered slow?
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1. To hear, three of the tiniest bones in your body are stimulated. List them in the order that they
are stimulated by a sound wave. The vibrations are then passed to the fluid-filled coiled tube in the
inner ear called the __________________.
2. The back surface of the eyeball, called the ____________________, is covered in light sensitive
cells. Of these, the ____________identify color, whereas the ____________ work best in dim light.
Both send information to the brain via the ____________________.
3. Define perception. Define sensation. List 3 general senses and 5 special senses.
4. In the skin, ________________ respond to temperature, and _______________ respond to
pressure and vibration.
5. List the four basic categories of taste.
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Lab 8
The Senses
Background
Your sense organs (nose, eyes, ears, tongue, and skin) are continually taking in information and
sending it to the brain for processing. Each sense organ collects information about your external
environment and also detects changes within the body. Your sense organs start to work when
something stimulates special nerve cells called a sensory receptor in a sense organ. Once
stimulated, the receptors send nerve impulses along sensory nerves to the brain. Your brain then
tells you what the stimulus is and elicits a response. Receptors in the eye react to light, those in skin
sense touch, pressure, and temperature. Other receptors respond to pain or visceral stretch. Sensation is
the physical act of responding to stimuli and converting it to an electrical signal to be sent to the central
nervous system. Perception occurs primarily in the brain and is the process of organizing and
interpreting sensory information. One aspect of this is screening information so that you can focus on
whats important. The senses are organized into two groups. General senses include touch, pain, and
temperature, and receptors are widely distributed throughout the body. Special senses are associated
with complex structures in the head such as the eye and ear. Special senses are sight, taste, smell,
hearing, and equilibrium (balance).
Exercise 1 The Ears
How You Hear
When an object makes a noise, it sends vibrations (better known as sound waves) speeding through
the air. These vibrations are then funneled into your ear canal by your outer ear. As the vibrations
move into your middle ear, they hit your eardrum and cause it to vibrate as well. This sets off a
chain reaction of vibrations. Your eardrum, which is smaller and thinner than the nail on your pinky
finger, vibrates the three smallest bones in your body: first, the hammer (malleus), then the anvil
(incus), and finally, the stirrup (stapes). The stirrup passes the vibrations into a coiled tube in the
inner ear called the cochlea. The fluid-filled cochlea contains thousands of hair-like nerve endings
called stereocilia. When the stirrup causes the fluid in the cochlea to vibrate, the stereocilia move.
The stereocilia change the vibrations into messages that are sent to the brain via the auditory nerve.
The auditory nerve carries messages from 25,000 receptors in your ear to your brain. Your brain
then makes sense of the messages and tells you what sounds you are hearing.
How You Keep Your Balance
Near the top of the cochlea are three loops called the semi-circular canals. The canals are full of
liquid also. When you move your head, the liquid moves. It pushes against hair-like nerve endings,
which send messages to your brain. From these messages, your brain can tell whether or how your
body is moving. If you have ever felt dizzy after having spun around on a carnival ride, it was
probably because the liquid inside the semicircular canals swirled around inside your ears. This
makes the hairs of the sensory cells bend in all different directions, so the cells' signals confuse your
brain.
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The Rinne Test for Conduction Deafness

1) Make a tuning fork vibrate by striking it with a rubber mallet.
2) Place the handle of the tuning fork against the bone behind your ear.
3) When the sound is no longer heard, bring the tuning fork around and hold it in front of the
opening to the ear.
4) If there has been no damage to the eardrum or ear bones, the sound should reappear.
Question 1. Record the results of the Rinne test.
Results for right ear_____________
Results for left ear______________
Question 2. What is the difference between conduction deafness and sensory-neural deafness?
(you are going to have to look this one up).
Question 3. Why do you think people with conduction deafness wear hearing aids that sit
against the bone behind their ear and not in the ear canal?
Question 4. How could you use the tuning fork to test for sensory-neural deafness?
Question 5. Why do you think people with sensory-neural deafness cannot be helped with a
hearing aid?
Test your sense of balance
1) Stand upright with your partner nearby in case you fall.
2) Have your partner record the length and quality of your attempts in Table 8.1.
3) Bend your right leg and hold your foot in your right hand.
4) Balance like this as long as you can (minimum 30 secs).
5) Try the same pose, but close both of your eyes.
6) Again, have your partner nearby recording your attempts.
7) Balance for as long as possible.
8) Try the same pose, but close your eyes and tilt your head back slowly.
9) Again, have your partner nearby recording your attempts.
10) Balance for as long as possible.
Question 6. Which stance made it the hardest to balance? Why do you think that is?
Question 7. How much of an impact does your inner ear have on your sense of balance?
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Exercise 2 The Eyes

How Your Eyes Work
Take a moment to locate an object around you.
Do you know how you are able to see it?
Would you believe that what you are actually
seeing are beams of light bouncing off of the
object and into your eyes? It is hard to believe,
but it is true. The light rays enter the eye
through the cornea, which is a thick,
transparent protective layer on the surface of
your eye. Then the light rays pass through the
pupil (the dark circle in the center of your eye)
and into the lens. When light rays pass through
your pupil, the muscle called the iris (colored
ring) makes the size of the pupil change
depending on the amount of light that's
available. You may have noticed this with your
own eye if you have looked at it closely in a
mirror. If there is too much light, your pupil will shrink to limit the number of light rays that enter.
Likewise, if there is very little light available, the pupil will enlarge to let in as many light rays as it
can. Just behind the pupil is the lens and it focuses the image through a jelly-like substance called
the vitreous humor onto the back surface of the eyeball, called the retina. The retina, which is the
size of your thumbnail, is filled with approximately 150 million light-sensitive cells called rods and
cones. Rods identify shapes and work best in dim light. Cones on the other hand, identify color and
work best in bright light. Both of these types of cells then send the information to the brain by way
of the optic nerve. When they send the image to the brain, the image is upside down. It is the
brain's job to turn the image right-side up and then tell you what you are looking at. The brain does
this in a specific place called the visual cortex.
Protection
Because the eye is such an important and complex part of our body, we have many features which
protect the eye. The eyebrows are the strips of hair above your eyes which prevent sweat from
running into them. Eyelashes help keep the eye clean by collecting small dirt and dust particles
floating through the air. The eyelashes also protect the eye from the sun's and other light's glare.
The eyelids sweep dirt from the surface of the eye. The eyelid also protects the eye from injury.
Tears are sterile drops of clean water which constantly bathe the front of the eye, keeping it clean
and moist.
Imperfect Eyesight
Not all people have perfect vision. People who can see things up close, but not far away are
considered to be nearsighted. This happens when the light entering the eye focuses on a point in
front of the retina. It occurs when the eyeball is too long or the cornea is too round, so the lens
cannot flatten enough to compensate. On the other hand, people who can see far away objects but
not those that are up close are farsighted. Farsightedness occurs when the light that enters the eye
focuses on a point behind the retina. This occurs when the eyeball is shorter than normal, causing
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the lens to focus images behind the retina. Whether a person is nearsighted or farsighted, glasses or
contacts help that person to see things much more clearly.
Two eyes are better than one, especially when it comes to depth perception. Depth perception is the
ability to judge objects that are nearer or farther than others. To demonstrate the difference of using
one eye versus two to judge depth complete the following:
Depth Perception
1) Hold the ends of a pencil/pen in each hand, hold them vertically or horizontally facing each
other at arms length from your body.
2) Now, close one eye and try to touch the ends of the pencils together.
3) Now try with two eyes: it should be much easier. It is easier with two eyes because each eye
looks at the image from a different angle.
Why do you need two eyes?
1) With your arms fully extended, hold a plastic drinking straw in one hand and a pipe cleaner in the
other.
2) With both eyes open, try to insert the pipe cleaner into the straw.
3) Now close your right eye. Try to insert the pipe cleaner into the straw. Repeat step c, but this
time close your left eye instead.
Question 8. How does closing one eye affect the ability to judge distances?
1. Test your visual acuity using the eye chart.
Visual acuity is usually measured with a Snellen chart. The Snellen chart displays letters of
progressively smaller size. "Normal" vision is 20/20. This means that the test subject sees the same
line of letters at 20 feet that a typical person sees at 20 feet. 20/40 vision means that the test subject
sees at 20 feet what an average person sees at 40 feet. Another way of saying this is that a person
with 20/40 vision has vision that is only half as good as the average, or, objects must be at half the
normal distance for her/him to see them. A person with 20/20 vision is able to see letters 1/10th as
large as someone with 20/200 vision. However, 20/15 vision is better than 20/20. A person with
20/15 vision can see objects at 20 feet that a person with 20/20 vision can only see at 15 feet.
1) Stand 20 feet away from the chart.
2) Cover one eye with your hand (dont press on your covered eye or it will be blurry when you
switch). If you have glasses, do one reading with glasses and one without glasses.
3) Have your partner point to one line at a time while you call out the letters.
4) Record your results in Table 8.2 and then switch and do the other eye.
Note that the set of numbers to the side of the row of letters always starts with 20. This number
simply corresponds to the number of feet you are standing from the chart. The second number refers
to the distance that a person with normal vision would be standing from the chart if that were the
persons last correct line.
Question 9. How would you describe the eyes of the person for which the above readings were
taken?
Question 10. Is the person nearsighted or farsighted or neither?
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Test the age of your eyes

1) Hold a pencil or ball point pen vertically at arms length.
2) Close your left eye and focus on the tip.
3) Quickly bring the pencil closer to your eye until it is out of focus.
4) Have your partner measure the distance (in centimeters) between your eye and the pencil.
5) Repeat for both eyes.
6) Try it with and without glasses (if you wear glasses).
Age of your Eyes

CM
9
Age
10
10
20
13
30
18
40
50
50
83
60
Question 11. What was the age of the test subjects eyes?
Right w/o glasses ____________ w/glasses____________
Left w/o glasses ______________ w/glasses____________
Question 12. How does this compare to your actual age?
Question 13. Are the right and the left eye the same age? If not, why do you think that could
be?
Find your blind spot

1) Obtain the optic disk card and hold it at arms length.
2) Hold it with the cross on the right and the dot on the left.
3) Close your right eye and stare at the cross with your left eye.
4) Slowly bring the card closer to your face until the dot disappears and then reappears again.
Question 14. Explain why the dot on the optic disk card disappears. (Hint: Look at the
diagram of the eye. How do impulses get to the brain?)
Observe the Shimmering Image

1) The image below appears to shimmer due to small imperceptible motions of your eyes.
Normally your brain will edit out the motion, but in this case, it does not.
2) Move your head slightly from side to side while looking at it to increase the shimmering.
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Test: is the image below moving?
After Images
Afterimages are a visual illusion caused by the relatively slow biochemistry of the
phototransduction process in the retina. Light interacts with rhodopsin in photoreceptor cells.
Rhodopsin is composed of a protein and a chromophore, a chemical that can absorb light of certain
wavelengths. When exposed to light, the bond structure of the rhodopsin molecule changes and the
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protein and chromophore dissociate. Cells in the retina enzymatically regenerate the rhodopsin,
but in strong light the rhodopsin can be used faster than it can be regenerated.
1) Stare at the red dot (between her eyes) in the picture for 20 seconds.
2) Now look at a white wall/ceiling for 10 seconds or just close your eyes for 10 seconds.
Question 15. Describe what you see when you close your eyes.
(movement, flickering, etc).
What do you notice?
Find the hole in your hand

1) Obtain a paper tube.
2) Hold the tube up to one eye.
3) Look at an object across the room through the tube.
4) Keeping both eyes open, bring your hand up next to the tube in front of your free eye.
5) Move your hand towards and away from your face until a hole appears in your hand.
6) Try it again with the other eye looking through the tube.
Question 16. Why does it appear that you have a hole in your hand?
Question 17. Does it work better with one eye or the other? Which one?
Circles or Ovals
1) Obtain a slightly flattened paper tube.
2) Hold one of the round tubes up to one eye and the tube that you flattened up to the other eye.
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3) Look through the tubes at the white screen, wall, or paper. Overlap the spots. Do you see the
circle or the oval?
4) Switch the tubes and repeat. If you saw only the circle before, you may see the oval now.
5) Your eyes and brain have trouble merging the different shapes. Most people have a dominant
eye. The brain will choose to see the image that is coming from the dominant eye. Some people do
not have a dominant eye, and therefore see the two shapes overlapped.
Question 18. After completing the Circle or Oval exercise, do you think you have a
dominant eye? Explain.
Question 19. Is it the same eye as the one you answered for #16 above? Why?
Testing for Color Blindness
People who are colorblind cannot see certain colors. There are many different types of color blindness,
but red-green color blindness is the most common. There are color vision tests that can assess if a person
has normal color vision or not. Ishihara plates are a common test for color blindness.
Question 20: What are the numbers of the Ishihara plates above, from left to right?
I Thought Spelling Was Important
1) Read the sentence below quickly.
I cdnuolt blveiee taht I cluod aulaclty uesdnatnrd waht I was rdgnieg. The
phaonmneal pweor of the hmuan mnid Aoccdrnig to a rscheearch at Cmabrigde
Uinervtisy, it deosn't mttaer inwaht oredr the ltteers in a wrod are, the olny
iprmoatnt tihng is that the frist and lsat ltteer be in the rghit pclae. The rset can
be a taotl mses and you can sitll raed it wouthit a porbelm. Tihs is bcuseae the
huamn mnid deos not raed ervey lteter by istlef, but the wrod as a wlohe.
Amzanig huh? Yaeh and I awlyas thought slpeling was ipmorantt!
Question 21.
Write out a message (at least two sentences) like the paragraph above,
discussing the structure or function of the eye:
Question 22. Was it harder to read or write with bad spelling? Why do you suppose that is?
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Exercise 3 Touch
While your other four senses (sight, hearing, smell, and taste) are located in specific parts of the
body, your sense of touch is found all over. This is because your sense of touch originates in the
bottom layer of your skin called the dermis. The dermis is filled with many tiny nerve endings
which give you information about the things with which your body comes in contact. They do this
by carrying the information to the spinal cord, which sends messages to the brain where the feeling
is registered.
The skin has numerous mechanoreceptors that
respond to pressure and vibration and
thermoreceptors that respond to temperature.
When adequately stimulated, the receptors
convert the mechanical or heat stimuli into
electrical signals that travel along axons to the
brain or spinal cord for interpretation. Some
areas of the skin are more sensitive to touch than
others. The level of sensitivity depends on
receptor density and the size of the receptive
field. The receptive field is the area where a
stimulus will cause a neuron to fire. For
example,
the
receptive
field
of
a
mechanoreceptor in the skin is the patch of skin
that particular neuron responds to. Large
receptive fields allow a neuron (receptor) to
sense changes over a large area but without
much precision. In the most sensitive areas of
the body, there is a high density of
mechanoreceptors with small receptive fields.
The two-point discrimination test reveals the ability of the somatosensory system to determine if the
skin is being touched in one or two points. If the two points are in an area of skin that has a low
touch receptor density and large receptive fields, only one point may be perceived. If the two points
are in an area of skin that has a high touch receptor density and small receptive fields, the subject
may perceive those two points. That is, two stimuli are perceived as separate if they are stimulating
the receptive fields of two different mechanoreceptors.
Two-point discrimination test
1. Your partner, the subject of the experiment, should be seated comfortably, eyes closed.
2. Begin with the two ends of the toothpicks together. Gently (lightly touch the skin, do not poke the
subject), place the tips of the toothpicks to the skin at the same time.
3. Your partner, the subject, should report feeling one or two points.
4. Move the points 1 mm further apart and gently touch the tips of the toothpicks to the skin.
5. Repeat steps 3-4 until the subject reports feeling two points.
6. Record the smallest separation distance (between the two points) that the subject reports feeling
two points.
7. Repeat this procedure on the different skin regions listed and record your data in Table 8.3.
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Question 23. What can you conclude about the density of touch receptors in your skin?
Adaptation is the decline in frequency of nerve impulses (in a sensory neuron) even when a receptor is
stimulated continuously and without a change in stimulus strength. That is, adaptation is occurring when
a sensory signal decreases in the presence of an above-threshold, persistent stimulus. Adaptation is part
of many sensory systems. For example, when we walk into a house before a holiday feast, the smell of
the food is immediate and intense. After a few minutes we no longer sense these odors at the same
intensity. Adaptation allows the nervous system to disregard continuous sensory information and be
prepared for changing or new incoming sensory stimuli.
1) Have your partner (his/her eyes can be open for this experiment) place the right hand in the ice
water bath and the left hand in the hot water bath.
2) Record your partner's immediate perceptions.
3) Wait two minutes. Record your partner's perceptions at this time. Did one hand adapt more
quickly than the other? (If this is too painful, do not keep your hand in the cold water for 2
minutes).
4) Have your partner remove his/her left hand from the warm water and place it into the room
temperature water. Record your partner's immediate perceptions.
5) Have your partner remove his/her right hand from the cold water and place it into the room
temperature water. Record your partner's immediate perceptions.
Question 24. What is the temperature of water according to your partners left hand?
Question 25. What is the temperature of water according to your partners right hand?
Question 26. Explain the above results. Cant you trust your senses?
Exercise 4 Smell
Olfaction refers to the sense of smell, which has similarities in all terrestrial and many aquatic
vertebrates. The mechanisms that control olfaction are divided into distinct regions. The olfactory
epithelium, located in the roof of each nasal cavity in humans, is the organ of smell. Special
olfactory receptor cells, numbering about 25 million, make up the bulk of this epithelium. Smells, in
the form of individual molecules, are bound to receptor molecules in the membranes of the cilia that
extend into the fluid mucus layer that coats the epithelium. The membranes of the nose secrete these
fluids to keep the molecules in solution to facilitate smell. Axons from the receptor cells lead to and
synapse with the olfactory bulb that lies just under the frontal lobe of the brain. Special cells in the
bulb are activated by smell stimuli and carry impulses toward the olfactory tubercle, where the
impulse is sent to the limbic system, thalamus, and cortex. In evolutionary terms, the limbic system
is one of the oldest parts of the brain. It contains such structures as the hippocampus, amygdala,
fornix, and mammillary bodies, and is extremely important to the emotional aspects of our
experiences. This fact explains why smells often evoke emotional memories. Other fibers terminate
in the olfactory cortex at the front of the cerebrum and are interpreted as the tens of thousands of
chemical scents that humans are capable of identifying.
1) Have the subject close his/her eyes, and distinguish the odors of (a) oil of cloves, (b) oil of
peppermint, and (c) tincture of camphor, when vials of these substances are held close to his/her
nose.
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Question 27. Did the test subject correctly determine each of the odors? Did any of the odors
remind him/her of anything?
2) Close one nostril and have the subject smell the tincture of camphor until the odor can no longer
be detected. Immediately have the subject attempt to distinguish, with the adapted nostril, between
oil of cloves and oil of peppermint. Record your results in Table 8.4.
Question 28. Describe your results.
Question 29. Describe why this happens.
Exercise 5 Taste
Have you ever thought about why foods taste different? It's really quite amazing. Your tongue and
the roof of your mouth are covered with thousands of tiny taste buds. When you eat something, the
saliva in your mouth helps break down your food. This causes the receptor cells located in your
tastes buds to send messages through sensory nerves to your brain. Your brain then tells you what
flavors you are tasting.
Taste buds probably play the most important part in helping you enjoy the many flavors of food.
Your taste buds can recognize four basic kinds of tastes: sweet, salty, sour, and bitter. The
salty/sweet taste buds are located near the front of your tongue; the sour taste buds line the sides of
your tongue; and the bitter taste buds are found at the very back of your tongue. Everyone's tastes
are different. In fact, your tastes will change as you get older. When you were a baby, you had taste
buds, not only on your tongue, but on the sides and roof of your mouth. This means you were very
sensitive to different foods. As you grew, the taste buds began to disappear from the sides and roof
of your mouth, leaving taste buds mostly on your tongue. As you get older, your taste buds will
become even less sensitive, so you will be more likely to eat foods that you thought were too strong
as a child. What if you could not taste anything? Things like medications, smoking, not getting
enough of the right vitamins, injury to the head, brain tumors, chemical exposure, and the effects of
radiation can cause taste disorders.
Location and distribution of various types of taste receptors
1) To locate and study the distribution of the taste receptors, place approximately 1 ml or less of the
10% salt solution in a watch glass, dip a cotton tipped applicator into the solution, and apply to the
surface of the tongue. Apply to right side, left side, tip, center and back of tongue. Properly discard
applicator immediately after use. Care must be exercised to be sure that all taste buds are
stimulated in the same manner. Avoid use of excess solution on the surface of the tongue. Note the
areas on the tongue where the sensation of salt can be tasted; carefully record those areas where the
sensation is most acute in Table 8.5.
2) Repeat for each of the following: 5% glucose, 1% acetic acid, and 0.1% quinine sulfate.
Sketch 1. Draw and label where on your tongue, you tasted each of the following:
SALTY
BITTER
SWEET
SOUR
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Question 30. How do your results compare to your partners?

3) Place some crystals of sugar on the tongue after you have wiped it dry with a piece of gauze.
Question 31. Do you taste anything? Why is it necessary for a substance to be in solution
before it can be tasted?
Exercise 6 Reflexes
Reflexes are involuntary, instantaneous movements in response to stimuli. Reflexes are mediated
via a reflex arc, which includes a receptor, sensory neuron, integration center, motor neuron,
and effector. A stretch reflex is a muscle contraction in response to stretching within a muscle.
Patellar Reflex
The patellar (knee-jerk) reflex is an example of a stretch reflex. The patellar reflex tests the
conduction of the femoral nerve.
1) Sit with your legs crossed or in a seat high enough to leave your legs dangling. Relax your legs.
2) Have your lab partner tap the patellar ligament with the blunt side of a patellar reflex hammer.
The tap should be just below the kneecap, and firm, but not hard enough to hurt.
3) Look for an extension of the leg as a response to the patellar reflex.
4) Check the reflex movement of your foot:
Normal reflex foot moves a few inches
Hyperreflexic foot moves extensively
Hyporeflexic foot moves little or not at all
Question 32. Describe the patellar reflex. Was it strong?
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Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
Question 11
Question 12
106
Question 13
Question 14
Question 15
Question 16
Question 17
Question 18
Question 19
Question 20
Question 21
Question 22
Question 23
107
Question 24
Question 25
Question 26
Question 27
Question 28
Question 29
Question 30
Question 31
Question 32
Table 8.1 Balance

Stance
Standing on one foot
One foot, eyes closed
One foot, eyes closed, head
back
Time (min)
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Stability
high---moderate---low
Table 8.2 Visual acuity

Right eye
Left eye
Without glasses
20/
20/
Table 8.3 Touch Receptors

Location
Distance when two points are
felt
Fingertip
Heel of hand
Forearm
Elbow
Back of neck
With glasses
20/
20/
Rank the concentration of receptors

(high:1, low: 5)
Table 8.4 Olfactory Adaptation Data

Substance
Sensed alone?
Adapt to camphor, distinguish

cloves/peppermint?
Oil of clove
Oil of peppermint
Tincture of camphor
Table 8.5 Taste Modalities Data
Substance
Taste Sensation Most Acute
Sucrose, 5%
Sodium Chloride, 10%
Acetic Acid, 1%
Quinine Sulfate, 0.1%
Sketch 1
(sketch below)
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Post-lab Homework 8
questions
1) Describe in detail how you hear a sound.

2) What types of sensory receptors are in the skin? What are their functions?
3) What is the anatomical basis for the blind spot of the retina?
4) Explain the functional, adaptive reason that receptive field and receptor density would differ
from place to place in the body? What is the area of greatest sensitivity and why is it so sensitive?
What is the area of least sensitivity and why isn't it more sensitive?
5) Describe the advantages and disadvantages of temperature adaptation. Do other types of
receptors adapt? Give illustrations.
6) What structures in the eye affect visual acuity? Describe the functions of each.
7. Why is the Eustachian tube important?
8. What does 20/20 vision mean?
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1. How many bones are in the human body? The bones of the girdles and limbs make up the
___________________ whereas the skull, vertebral column, ribs and the sternum make up the
___________________.
2. Which kind of bone is dense, smooth and homongenous? Which kind of bone is made of small,
needlelike bars and has lots of open space? For the following bones, indicate if the bone is long,
short, flat, sesamoid, or irregular.
a. Humerus
b. Rib
c. Pelvic girdle
d. Knee cap
e. Femur
3. Thick filaments are made up of the protein ___________________ and thin filaments are made
of the protein ________________________.
4. What are the components that make up a sarcomere?
5. What is rigor mortis? Why does it sometimes occur?
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Lab 9
Muscles and Skeleton
Background
All movement in higher animals is dependent upon the contractile capabilities of muscle. But
peristaltic contractions of the intestine differ in duration and nature from the muscular contractions
which flex your arm, and different types of muscle are used to accomplish each. The three types of
muscle found in vertebrates are: (1) smooth, (2) skeletal, and (3) cardiac. All three muscle tissues
are composites comprised of (1) muscle cells, (2) connective tissue, (3) nerve fibers, and (4)
blood vessels. The muscle cells are wrapped by connective tissue which maintains the shape of the
muscle, and carries the blood vessels and nerves to the muscle tissue. The muscle cells themselves
are elongate, and are often referred to as muscle fibers. These contain the intracellular contractile
elements, the myofilaments. The intracellular organization of these elements is greatest in skeletal
muscle and least in smooth muscle.
The skeletal system consists of bones and
ligaments. In embryos, the skeleton is composed
mainly of cartilage, but in adults, most of the
cartilage is replaced by rigid bone. Bones
support the body, protect soft organs, and
provide leverage for the skeletal muscles of the
body. In addition, bones store lipids and
minerals. The skeleton is made up of bones that
are connected at joints, and is subdivided into
two divisions: the axial skeleton (skull, the
vertebral column, the ribs and the sternum;
shown in green), and the appendicular skeleton
(bones of the girdles and limbs; shown in
purple). There are 206 bones in the human body
and they are all composed of two basic kinds of
tissue that differ in texture: compact bone is
dense and looks smooth and homogenous,
spongy bone is composed of small, needlelike
bars of bone and a lot of open space.
Human bones come in different sizes and
shapes. Bones can be classified on the basis of
shape. Common bone shapes include long
bones, short bones, flat bones, and irregular
bones. Long bones are longer than they are
wide. Long bones include all bones of the limbs,
except the patella (kneecap), wrist bones, and ankle bones. Short bones are smaller than long
bones and many are roughly cube-shaped. Short bones include the wrist bone and ankle bones. A
sesamoid bone is a type of short bone that is shaped like a sesame seed and form in a tendon. The
patella (kneecap) is an example of a sesamoid bone. Flat bones are flattened bones. Flat bones
include the sternum (breastbone), scapulae (shoulder blades), ribs, and most skull bones. Irregular
112
bones are irregularly shaped and do not fit into any of the preceding categories. Irregular bones
include the vertebrae and hip bones.
Exercise 1 Muscle Tissue
(adapted from Carolina Biological)
Muscle tissue is made of fibers formed by the fusion of cells during development. A single muscle
fiber, barely visible to the unaided eye, has many nuclei that lie close to its outer membrane. Each
fiber contains hundreds of long, threadlike structures called myofibrils, arranged in parallel. About
75% of a muscles total volume is made up of myofibrils. Myofibrils are the structures that carry
out muscle contraction. Under a microscope myofibers look striated (striped), with a repeating
pattern of bands and lines perpendicular to the length of the fiber. The banded pattern is caused by
an organized, parallel arrangement of protein filaments within the myofibrils. There are two types
of filaments in a myofibril: thick filaments composed of the protein myosin (shown in purple in
the bottom portion of the figure below), and thin filaments composed of the protein actin (shown
in pink in the bottom part of the figure below). The filaments overlap in an orderly, repeated
manner, creating units called sarcomeres. When many filaments are bundled in a cylinder, the
repeated overlapping pattern of filaments results in the banded pattern seen under the microscope.
When you observe the glycerinated muscle fibers with a compound microscope, you should be able
to see bands.
Muscle contraction occurs through the interaction of the actin and myosin filaments in the
sarcomeres. When a muscle contracts, the myosin binds to the actin filaments in a manner that
causes the actin filaments to be pulled together. For a muscle fiber to contract, the myosin heads
113
must first be activated by ATP. One molecule of ATP binds to a myosin head and is hydrolyzed to
ADP and inorganic phosphate (Pi). Both ADP and Pi remain bound to the myosin head, and the
energy released from ATP hydrolysis is transferred to the myosin head as well. The myosin head is
now activated. Imagine holding one end of a thin, plastic ruler and pulling back on the other end so
that the ruler bends. When you release the ruler, it will spring back to its original shape. The
activated myosin head is rather like the bent ruler. When the myosin head binds to the actin
filament, its energy is released and the myosin head springs back, carrying the bound actin filament
with it. This movement causes the muscle fiber to contract. After the myosin head has sprung, it can
interact with a new molecule of ATP. When ATP binds to the myosin head, it releases the actin
fiber, ADP, and Pi . The myosin head is now reactivated, and the cycle can begin again.
If no ATP is available to reactivate the myosin, the actin/myosin complex remains locked together,
and the muscle cannot relax. When an animal dies, its cellular ATP stores are depleted and all its
muscles lock. This locked condition is called rigor mortis. In living animals, muscles resume their
normal shapes (relax) after contraction because they are pulled by opposing muscles.
The just-described picture of contraction in living muscle is incomplete because it omits the role of
nerve signals in instigating contractions. Muscle fibers do not normally contract without appropriate
nerve signals because a control mechanism prevents them from doing so. The control mechanism
works through two regulatory proteins, tropomyosin and troponin, which form a complex lying
along the actin filament. The troponin/tropomyosin complex blocks the myosin binding site on the
actin fiber, preventing myosin from binding to actin and causing contraction (Figure 5a). The signal
for muscle contraction is a nerve impulse to the muscle fiber that results in intracellular release of
calcium ions. The calcium ions induce contraction by binding to troponin. When Ca2+ binds to
troponin, the shape of the troponin/tropomyosin complex is altered. This change in configuration
allows activated myosin crossbridges to bind to the actin and release their energy as motion
Therefore, in a normal muscle fiber rich in ATP, the myosin heads are activated and ready to cause
a contraction, but they cannot until a nerve impulse releases Ca2+. The contraction cycles will
continue as long as intracellular Ca2+ concentration is high and as long as ATP is available. When
Ca2+ levels fall, Ca2+ is released from the troponin molecules, and the troponin/tropomyosin
complex again blocks the binding sites on the actin fiber.
The glycerinated muscle system is different from muscle in living tissue. The glycerination process
removes ions and ATP from the tissue and disrupts the troponin/tropomyosin complex so that the
binding sites on the actin fibers are no longer blocked. No Ca2+ is needed to induce contraction.
However, no ATP is present in the glycerinated tissue, so the myosin heads are not activated. Mg
ions have been reported to interfere with muscle contraction in live tissue, but in glycerinated
tissues, the addition of KCl and MgCl2 solutions to ATP increases the strength of muscle
contraction because of myosin's high affinity for these ions (they make myosin more activated). In
this experiment, you will be experimenting with adding ATP and ions to the glycerinated tissue to
initiate contraction. When contraction occurs, you will be able to see the change in length of the
sarcomeres and measure the overall shortening in the length of the dissected muscle tissue. After the
muscle is contracted it will not relax, because there is no opposing muscle to stretch it out.
114
Materials
- glycerinated skeletal muscle strips in a 50% glycerol solution
- 0.25% ATP in distilled water
- 0.25% ATP plus 0.05 M KCl plus 0.001 M MgCl2 in distilled water
- 0.05 M KCl plus 0.001 M MgCl2 in distilled water
-magnifying glasses
-microscope slides
-pipets
-millimeter ruler or micrometers
-sharp scissors
-dissecting needles
-dissecting and compound microscopes
1. On your lab table you will find a petri dish containing a bundle of mammalian skeletal muscle
fibers. Place the petri dish containing a segment of skeletal muscle tissue on the stage of a
dissecting microscope. If a microscope is unavailable, you may use the supplied magnifying glass.
Use dissecting needles to gently tease the segment into very thin strands (about the size of a strand
of silk). You will see optimal results with single muscle fibers, but these are difficult to obtain. The
thinnest strand that you will likely get is a group of two to four fibers. Strands of muscle that are too
thick will not work in this exercise.
2). Place the strand on a microscope slide do not cover with a coverslip. Do not let the fiber dry
out. Pipette glycerol from the petri dish if needed. Observe under low power. Notice the striations
of the myofibers.
Sketch 1 - 2. Sketch your muscle fiber and label recognizable parts.
2. You will probably notice that you have more than one fiber. Continue separating fibers until you
have a single fiber. Transfer your extra fibers to clean microscope slides, so that you have a total of
three slides with single fibers on them. Be sure to arrange the fibers out straight, and keep them
moist with glycerol.
3. Measure the length of each fiber in millimeters and record these lengths in Table 9.1.
4. Using your first slide, and while observing through the microscope, add one drop of 0.25% ATP
solution containing 0.05 M KCl and 0.001 M MgCl2. Watch for 30 seconds.
Question 1. What happened? Have the fibers changed length, width, or both?
Re-measure length and record in Table 9.1.
Question 2. Examine under a compound microscope and describe the striations.
5. Repeat Step 4 by adding a second drop of 0.25% ATP solution containing KCl and MgCl2. Remeasure the length and record in Table 9.1.
115
Question 3. What effect does a second drop of the ATP-salt solution have on muscle
contraction?
6. Get your second slide and repeat Step 4 using a drop of ATP only solution.
Question 4. What happens? Re-measure and record your data in Table 9.1.
7. Get your third slide and repeat Step 4 using a drop of KCl plus MgCl2 only solution.
Question 5. What happens? Re-measure and record your data in Table 9.1.
Question 6. Why was the ATP solution needed?
Question 7. Why do you think the KCl + MgCl2 solution was needed?
Question 8. How does the contracted fiber look different from the uncontracted fibers?
Question 9. Why do the muscle strands remain contracted permanently after adding the ATP
solutions?
Exercise 2 Bone
1) Examine the slide of a cross section through bone. The functional units of compact bones are
called Haversian Systems. A Haversian system consists of the central (Haversian) canal, the
cavity in the center where blood vessels and nerves are found, the lacunae or little cavities where
the mature bone cells called osteocytes are found, and the canaliculi or little canals that allow
communications between osteocytes in neighboring lacunae. The lacunae are arranged in concentric
circles or lamellae, and the substance in between is primarily salts of calcium and phosphorus in
addition to collagen fibers.
116
Sketch 3. From your slide, sketch a Haversian system and label the Haversian canal,
canaliculi, lacuna, and osteocyte.
Question 10. Why is bone considered a type of connective tissue?
Like other connective tissues, bone consists of a matrix containing fibers (primarily collagen) and
cells embedded in a ground substance. However, in bone the ground substance is in large part
calcium salts; in fact, the major portion of the dry weight of bone may be composed of these
inorganic salts. This makes the matrix of bone hard, so that it is an ideal tissue to serve as a rigid
support for the body. Because bone is hard and we will study only dried bone, it is easy to think of
bone as inert, like concrete. However, it is important to realize that bone is a living tissue which
contains living cells.
2) View the unidentified bones on display. Determine what type of bone (long, short, flat, irregular)
each unknown is, and determine if each is part of the axial or appendicular skeleton. Fill out Table
9.2 with your results.
117
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
Question 10
118
Sketch 1
Sketch 2
Table 9.1 Length of Glycerinated Skeletal Muscle Following Exposure to Different Solutions
Length without Length after ATP Length after
Length after
exposure
+ KCl + MgCl2
ATP exposure
KCl + MgCl2
N/A
N/A
Fiber 1
N/A
N/A
Fiber 2
N/A
N/A
Fiber 3
Sketch 3
Table 9.2 Identification of Unknown Bones

Type of Bone
A
B
C
D
E
F
G
Skeletal System
119
Post lab 9
questions
1. Describe the roles of nerves, muscles and bones in producing movement.
2. Explain the sliding filament theory of muscle contraction.
3. How is skeletal muscle like cardiac muscle? How do they differ?
4. List four basic functions of the skeletal system.
5. Describe the role of Ca++ in muscle contraction.
6. Sketch and label a sarcomere.
7. Sketch and label a neuromuscular junction.
8. Name 3 bones in the human body, describe function, and identify each as either axial or
appendicular.
120
Pre-lab Week 10
1. In humans, sperm is deposited in the _____________ where it must quickly move to the
______________ and then to the ________________ where it can survive for a few days. Normal
fertilization takes place in the ___________________.
2. What does hyperactivate mean in reference to sperm? Why is this important?
3. Briefly describe development from the zygote to the gastrula.
4. What will the ectoderm, mesoderm and endoterm tissues become in an adult?
5. Define blastocoels, blastopore, and archenteron.
121
Lab 10
Reproduction and Development
Background
Reproduction (by Jones, Gottlieb, and Suarez 2007)
In different species of mammals, males deposit sperm into the vagina (e.g., primates and cattle) or
directly into the uterus (e.g., pigs). Human sperm are deposited into the vagina right at the entrance
to the cervix, where thousands quickly enter. This saves the sperm from being killed in the vagina.
The pH of human vaginal fluid is highly acidic, which serves to kill bacteria and other potentially
infectious microbes; however, the acid can also immobilize sperm that fail to enter the cervix
quickly.
Sperm are able to swim through the cervix by following grooves in the wall. Although the cervix is
filled with mucus, the mucus is very watery during the fertile period of the month, particularly in
the grooves, and sperm can easily swim through the grooves to reach the uterine cavity. When
sperm reach the entrance to the oviduct, they must squeeze through the opening to get inside. Only
vigorously motile, well-shaped sperm can make it through. Sperm may be stored in the oviduct for a
few days (or a few months in hibernating bats) until ovulation occurs and the egg enters the oviduct.
When a human sperm is first deposited in the vagina, its flagellum
(tail) generates low symmetrical waves that propel it forward in a
straight path (see (A) in the Figure). This helps the sperm to pass
through the cervix and uterus, and to enter into the oviduct. When
the time of ovulation nears, the sperm in the oviduct are triggered
to hyperactivate. The waves generated by the flagellum increase
in height, but the beat becomes asymmetrical. If hyperactivated
sperm are put on a microscope slide, this asymmetrical beating
causes sperm to swim rapidly in zigzags or circles (see (B) in the
Figure)not the sort of behavior one would expect from a sperm that is about to fertilize an egg! A
microscope slide, however, is not the same as the inside of the oviduct. In the oviduct, only sperm
that are hyperactivated can reach the egg and fertilize it.
Some cases of human male infertility have been linked to poor hyperactivation of sperm. Sperm
hyperactivate when exposed to a trigger that raises calcium levels in the flagellum. The natural
trigger that hyperactivates sperm in the oviducts is still unknown, but sperm can be hyperactivated
by drugs that are known to increase intracellular calcium. One such drug is caffeine. Caffeine can
raise calcium levels by opening calcium channels in the plasma membrane of the sperm flagellum.
Exercise 1 Hyperactivation of Bull Sperm Using Caffeine
For this classroom activity, we will use caffeinated drinks to hyperactivate bull sperm. Bull sperm
behave quite similarly to human sperm, thus they serve as an excellent model to use for these
experiments. The main difference between bull sperm and human sperm is that bull sperm have
paddleshaped heads, while human sperm have conical heads.
Question 1. What do you think will happen when caffeine is added to sperm?
122
Materials
-Bull semen/milk solution (in clear tube)
-Eppendorf tubes (1.5 ml size)
-distilled water
-hot water bath
-plastic pipettes
-microscope slides and cover slips
-compound microscope
-caffeinated drink in milk solution (coffee) pH 8.3 (yellow tube)
-decaffeinated drink in milk solution (coffee) pH 8.3 (green tube)
-pure caffeine in milk solution pH 8.3 (purple tube)
Note: Sperm are diluted in milk rather than in water in order to approximate the fluids encountered
by sperm in the female reproductive tract. Adding plain water to sperm will kill them immediately.
Methods
Keep all tubes, pipets, slides, and coverslips warm (37 C). Sperm are delicate. It is important that
they are kept warm for the duration of the experiment and handled gently. Whenever transferring
sperm, pipet smoothly and slowly to avoid killing them. Slides must be very clean to avoid
contamination.
1) Warm milk and all test solutions to 37 C (98.6 F).
2) The sperm will work best if it is kept warm, treated gently, and protected from light as much as
possible. Perform all tests as soon as possible after warming sperm. Check to make sure that you
have all 4 tubes available to you: 1) 0.1 ml semen in 0.9 ml prewarmed milk , 2) caffeinated drink in
milk, 3) decaffeinated drink in milk, and pure caffeine in milk.
3) Stir up the sperm very gently by flicking the bottom of the tube. Using a plastic pipet, place a
small drop of the sperm suspension onto a microscope slide. Gently cover with a coverslip.
4) Quickly place the slide on the microscope stage and focus on the sperm using the 40X objective.
If you have trouble finding sperm, go to the edge of the coverslip and focus on the glass edge of the
coverslip. This will put you at the right focal length for seeing sperm. The sperm are 70 microns
long, so they are barely visible under a 10X objective. If the microscope is equipped with a substage
diaphragm in addition to the field diaphragm, the sperm will be easier to see if the diaphragm is
closed down to increase depth of field. Keep your condenser closed as much as possible to keep the
light down.
Question 2. Describe the swimming patterns of the sperm and estimate by eye the percentage
of sperm that are motile.
Record your data in Table 10.1.
123
4) Place a small drop of pure caffeine dissolved in milk solution on a clean slide. Immediately add a
small drop of sperm suspension into the drop of caffeine solution and top with a coverslip. Quickly
examine the sperm using the 40X objective on your microscope. Note your observations in Table
10.1.
Question 3. Describe the swimming patterns of the sperm and estimate by eye the percentage
of sperm that are motile.
Question 4. Compare the difference in movement between the sperm sample that was not
exposed to caffeine and the sperm sample that was.
Question 5. Compare the percentage of sperm moving the sperm sample that was not exposed
to caffeine and the sperm sample that was. Which sample has more sperm moving?
Hypothesize on why these samples may differ.
5) Repeat Step 4 using decaffeinated coffee in milk solution. Note your observations in Table 10.1.
6) Repeat Step 4 using caffeinated coffee in milk solution. Note your observations in Table 10.1.
Question 6. Compare the effect of decaffeinated coffee on sperm with that of caffeinated
coffee. Which sample has more sperm moving? Are the sperm moving in the same pattern in
both samples? Explain your result.
Question 7. Compare the effects of decaffeinated coffee on sperm with that of sperm that
were not exposed to any solutions. Describe similarities and/or differences.
Question 8. Describe the effects of the caffeine solution on sperm. Compare the results with
the sperm that was not exposed to solution, sperm exposed to coffee, and sperm exposed to
decaffeinated coffee. Is pure caffeine more effective? Does it kill more or fewer sperm?
Record any observations.
Question 9. What ingredients in the drinks might have killed sperm?
Caffeine is a drug that simulates whatever it is that actually hyperactivates sperm during the natural
fertilization process. The natural trigger for hyperactivation is yet to be discovered. Realistically,
very little caffeine from a cup of coffee would make it to the sperm in the reproductive tract. The
other organic chemicals in the drinks that kill sperm are unlikely to kill sperm in the body, because
most will be broken down by digestive enzymes. Caffeine is a small molecule that is not quickly
destroyed by digestive enzymes.
Exercise 2 Development
After an egg is penetrated by a sperm, the egg and sperm nuclei fuse to form a single nucleus. This
process is called fertilization. Fertilization is the initial event in development. The fertilized egg is
called a zygote. After production of the zygote, the embryo develops through a rapid cell division,
forming a hollow sphere of cells called the blastula. The blastula is the same size as the original
egg. This stage exhibits no cell growth but merely cell division, also called cleavage.
124
Materials
-Early embryonic stages slides of starfish
1) Using your starfish development slides, look first at the unfertilized egg using the 10X objective
of the compound microscope. Now focus in on the zygote. Note the distinctive fertilization
membrane. This membrane forms as soon as the sperm penetrates the egg.
Question 10. Why is the development of the fertilization membrane important?
2) Cleavage. The first identifiable period of embryonic development occurs as the fertilized egg
begins to divide. These early divisions are called cleavage divisions and each cell produced is a
blastomere. Focus on the starfish early cleavage stages. Compare the two cell, four cell and eight
cell stages.
Sketch 1: Sketch the two cell, four cell and eight cell stages of early cleavage.
Question 11. Is there a difference in size in any of these cell cleavage stages? Describe why
the cells are the size that you see.
3) Morula: continued division leads to a solid ball of cells called the morula around the 32-cell
stage. Focus on the starfish late cleavage stage.
Question 12. How does the morula compare to the eight cell stage? Are the nuclei still
visible?
Question 13. How does its size of the morula compare with that of the fertilized egg?
4) Blastula or Blastocyst: If the embryo is spherical it is called a blastula (nonmammal); if it is a
flattened disc it is called a blastocyst (mammal), both are equivalent stages. At the blastula stage,
the cells form a hollow sphere. The cavity inside is called the blastocoel.
Sketch 2: Sketch the blastula and identify the blastocoel
5) Gastrula Several hours after blastula formation, a second major morphological event occurs.
Some cells move inward and folding inward. This ingression and involution leads to the gastrula
with two layers of germ tissue. The outer layer of cells is referred to as the ectoderm which will
form the skin and nervous system. The infolding leads to an inner tube consisting of endoderm
which will form the digestive system. The blastocoel is still present, but now an additional space is
present the archenteron or space within the endoderm tube, which opens to the outside through
the blastopore. The archenteron will continue to grow until a second opening forms. This second
opening will become the mouth in the starfish and in vertebrates, while the blastopore will become
the anus. Near the future mouth, pouches will grow from the archenteron into the blastocoel. The
tissue forming these pouches will become the mesoderm, which will develop into muscle and
connective tissues along with some of the urinary and reproductive organs.
125
Focus on the gastrula and note the tube within the elongated embryo.
Sketch 3: Sketch the gastrula and label the ectoderm, endoderm, blastopore and archentron.
In the transition from blastula to gastrula, a lot has happened. The basic shape of an animal has
appeared. Many animals have a digestive tube-within-a-body-tube plan of organization, if you
ignore the appendages (compare an earthworm and a human). In addition, the three germ layers
have formed, each of which will develop into all of the adult tissues. Beyond the gastrula stage,
differentiation continues. Many animals, except for birds and mammals, have a free-living larval
stage. A larva is capable of feeding itself and continues to grow using external energy sources. At a
later stage, a metamorphosis takes place, and the larva changes into an adult. For the starfish, there
are two larval stages that appear as modified gastrula before a miniature adult with five arms
develops.
Be sure to show your answers, tables, and sketches to your TA before you leave today
126
Answer Sheet
laboratory
Question 1
Question 2
Question 3
Question 4
Question 5
Question 6
Question 7
Question 8
Question 9
127
Question 10
Question 11
Question 12
Question 13
Table 10.1 Effects of Caffeine on Sperm

Exposure
% of sperm still moving
Motion of sperm (straight line or

erratic)
Milk solution
Pure caffeine
Caffeinated coffee
Decaffeinated coffee
Sketch 1
Sketch 2
Sketch 3
128

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Biology for Allied Health

Portland State University

Pre-lab Questions Lab 4 ................................................................................................................. 39

Exercise 2 The Eyes ...................................................................................................................... 96

In-lab Homework (10-1 lowest=9) labs X 12 pts each

Post-lab Homework (9-1 lowest=8) labs X 8 pts each

Quizzes (9-1 lowest=8) quizzes X 10 pts each

Week 2 pre-lab Week

Tissues and the

Week 3 pre-lab Week

Nutrition and Digestion

Week 4 pre-lab Week

Week 5 pre-lab Week

Week 6 pre-lab Week

Endocrine and Nervous

Week 7 pre-lab Week

Week 8 pre-lab Week

Muscles and Skeletal

Important information: Please fill out on Day 1

Pre-lab Questions Lab 2

Exercise 2 Blood Pressure

Pulse Rate (beats/min)

Pre-lab Questions Lab 3

The part of the hair that is enclosed in a hair

Are they smooth and flat,

-prepared slide of adipose tissue

Question 21. In which part of the body does cartilage predominate?

Pre-lab Questions Lab 4

1. Describe the difference between mechanical and chemical digestion.

proteins (pepsin) polypeptides

starch (pancreatic amylase) maltose, lactose, sucrose

Exercise 1 Digestion of Proteins

4) Add 4 drops of hydrochloric acid to tube 4.

Exercise 2 Digestion of Carbohydrates

-Liquid soap (fat)

Table 4.1 Digestion of Macromolecules

1. Does the effectiveness of pepsin depend on pH? Explain your answer.

Pre-lab Questions Lab 5

units in the blood, such as blood cells and

Abnormal Urinary Components

Nitrite is a product of the bacterial reduction of nitrate and indicates the

Table 5.1 Presence/Absence of Glucose, Starch and Chloride Testing

Table 5.1 Presence/Absence of Glucose, Starch and Chloride Testing

Table 5.2 Three sample urinalysis

Pre-lab Questions Lab 6

1. Place the slide guide on a flat area in front of yourself.

3. Prepare to draw blood

a) What is Student A's blood type?

Exercise 2: Disease Transmission

2. As you investigate the slide of the spleen,

Table 6.1 Microorganisms and Disease Lab Data Sheet

Pre-lab Questions Lab 7

Table 7.1 Comparison of Hormonal Effects on Different Organs

Question 9. Explain the positive feedback loop observed in LH regulation.

Question 12. What are the effects of decreasing testosterone?

Figure 7.4. Graphic representation of organs studied in the autopsy.

Figure 7.1 Unlabeled Neuron

Pre-lab Questions Lab 8

The Rinne Test for Conduction Deafness

Exercise 2 The Eyes

Test the age of your eyes

Age of your Eyes

Find your blind spot

Observe the Shimmering Image