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Muscle

Physiology I
Formal Report
Herman
Bimikwe
Biology 282 L

Abstract:
In this experiment, our main objective was to analyze what happens during the process of muscle
contraction. To be more precise, we were to observe how the muscle contraction happens in the
presence of ATP. Using solutions of ATP and water, ATP with KCL + MgCl2, and KCl + MgCl2 without ATP.
We first preceded in making predictions of what would happen to the muscle fibers and then made
observations of what did happen and the recorded the fragments of change. The muscle tissue were
found to widen in the solution of ATP by an average of .1cm; shrink in the solution of ATP mixed with KCl
and MgCl2 by an average of .33cm, and no reaction in the last solution which was just KCl and MgCl2
mixed. This led us to conclude that ATP is the main drive of muscle contraction and that KCl and MgCl
are simply there as a complimentary enhancer but their solutions alone cannot trigger the process of
contraction.

Introduction:
The human body consists of 3 major types of muscles which are the smooth muscle, the
cardiac muscle and the skeletal muscles. Each of these muscles has specific functions and is
located in different parts of the human body. The cardiac muscles are located in the walls of the
heart and are described as involuntary muscles since we do not control them. The smooth
muscles are located in the walls of hollow visceral organs with the exception of the heart and
also happens to be part of the involuntary muscle category. The skeletal muscle is the ones
attached to the skeleton and they are described as voluntary since we possess full control of its
movement.

These muscles all have similarities and differences physiological wise. The cardiac and skeletal
muscles for example both appear striated whereas the smooth muscle as its name implies is
simply smooth and non-striated. What the smooth muscle have in common with the cardiac
muscle is that they both contain a single nucleus within each muscle sarcomere unit. The skeletal
muscle is unique in that I contains multiple nuclei due to cell fusion during development.
The smooth muscle is described as is due to the fact that it lacks striations that the other
muscles have. It produces longer lasting contractions due to no t-tubules compared to striated
muscles and they are mainly found in visceral organs such as the stomach, uterus, bladder,
intestine, and the walls of arteries. They possess a single nucleus and contain thick and thin
filaments but not patterned in a way that it would develop striations. The contraction of the
smooth muscle is regulated by sarcoplasmic (cytosolic) free Ca2+ concentration which rises
from resting levels of 120-270 nanometers to approximately 500-700 nanometers. This occurs in
response to stimuli like membrane depolarization, or adrenergic and muscarinic agonist. The
binding of Ca2+ to calmodulin activates myosin- light chain kinase, which as a response
phosphorylates myosin. This in return triggers the cycling of myosin cross bridges along actin
filaments and the development force. Protein Kinase C and Calponin may also play role in the
fine tuning of the contractile state. [Oxford Dictionary of Biochemistry and Molecular Biology]
All smooth muscles share many distinct characteristics. When relaxed the cells take the shape of
long, narrow spindles or worms. They also have the ability of contracting to about half or less of
their relaxed length. The main conductor of contraction is a rise in the cellular concentration of
calcium. This is usually triggered by a series of chemical signals that differ depending on the
type of smooth muscle involved which consist of neurotransmitters that are released at
autonomic nerve endings. The mechanism of calcium stimulation in smooth muscles differs from

striated muscles because it uses a different signaling protein called Calmodulin instead of
troponin. Another significant difference between smooth muscle and striated muscle is that it
uses metabolic energy more efficiently causing it to not become fatigued.
The skeletal muscle is the muscle in the body that is responsible for all voluntary
movements as well as automatic movement like moving our heads and standing up. On top of
being motors of the body, they are also used as breaks, shock absorbers, heaters (when
shivering), and protein storage when we face malnutrition. It contains a large amount of
myofibrils which consist of sarcomeres units also known as contractile units. Each of these
muscle fibers contains myofibrils which has the contractile apparatus and a system for
controlling the myofibrils through changes in calcium concentration. Each of the myofibrils runs
the whole length of the muscle fiber with a variable amount of sarcomere (muscle unit). The
myofibril also consist of myofilaments which are myosin, the thick filament which consist of a
single protein and thin filaments which contains actin.
Muscle contraction needs energy in order to conduct the cross-bridges through cyclic interactions
because myosin works in moving the thin filament. Energy is also used for the process of
calcium pumping by the sarcoplasmic reticulum. The energy required for muscle contraction
comes from ATP (Adenosine Triphosphate) to ADP (Adenosine diphosphate) and phosphate.
The skeletal muscle has enough ATP to power it at maximum output for just a few seconds.
Adenosine Triphosphate can also be regenerated in muscles quickly from phosphocreatine which
gives the muscle 10 to 20 more seconds of maximum activity.
The cardiac muscle is an involuntary muscle. It is the muscle abundant in the heart. Its
fibers are cylindrical and branched shaped allowing it to interlock for the contraction impulses to

take place. It also consist of a significant amount of mitochondria due to the large amount of O2
supply required by the heart. Another unique feature the cardiac muscle possesses is the
irregularly-spaced darks between myocytes known as intercalated discs. They are very prominent
where membranes of adjacent myocytes meet the functions of intercalated discs is to glue the
myocytes together so they dont pull apart during contraction and to allow an electrical
connection between the cells.
In this particular experiment, the muscle of choice was the psoas muscle of a rabbit. The
psoas muscle is a muscle in the groin that acts jointly with the iliacus muscle to flex the hip joint
[Oxford Reference] This muscle was ideal for this experiment because its fibers were long and
straight without the presence of connective tissues interfering it. It was stored in glycerin because
it is a substance that allows the muscle fibers to stay fresh retaining myosin, actin, troponin and
tropomyosin which are essential components for contraction.
So the purpose of this experiment was to make a careful analysis of the muscle contraction
occurring when placed in different solutions. Solution A consisted of .25% ATP in distilled
water, solution B consisted of a mixture of .25% ATP in water mixed with .05 M KCl + .001 M
MgCl2 and solution C was simply composed of .05 M KCl + .001 M MgCl2. Based on our basic
understanding of muscle contraction, we know that ATP is an indispensable factor in the process
of muscle contraction. So out of all tree solutions we hypothesized that that solution B will have
the most effective contraction due to the presence of ATP and the two complimentary enhancer
of contraction which are KCl and MgCl2.
Methods:

We began this experiment by obtaining a fresh piece of psoas muscle that had been stored in
glycerol to maintain its freshness. We got a small piece of the muscles measured at
approximately 2 cm and placed it in another solution of glycerol so that while preparing the vial
solutions it would remain intact. That muscle fiber was then divided (cut) into nine separate
strands to be tested in each solution vials. Solution A consisted of .25% ATP in distilled water,
solution B consisted of a mixture of .25% ATP in water mixed with .05 M KCl + .001 M MgCl2
and solution C was simply composed of .05 M KCl + .001 M MgCl2. Before proceeding further
we placed to muscle fibers under the microscope and measured them. After we added solution A
drop wise to the muscles and waited about forty-five seconds and placed it back under the
microscope and took measurement again to determine if the solution had any effect on the size
and other changes such as the color of the fiber. This same procedure was performed for solution
B and solution C respectively.
Results

Table
Treatment
(reagent applied
to muscle)

Fiber Length
(Before adding
reagent)

Fiber Length ( After


adding reagent)

Difference
in size

Color
changes

A: ATP only

1.5 cm

1.4 cm

0.1 cm

A: ATP only

2 cm

1.9 cm

0.1 cm

A: ATP only

1.3 cm

1.3 cm

No change

B: ATP + KCl
and MgCl2

1.4 cm

With two extra drops


1.0 cm

0.4 cm

B: ATP + KCl
and MgCl2

1.1 cm

With two extra drops


0.7 cm

0.4 cm

B: ATP + KCl
and MgCl2

0.7 cm

0.5 cm

0.2 cm

C: KCl and
MgCl2
C: KCl and
MgCl2
C: KCl and
MgCl2

1.5 cm

1.6 cm

1.7 cm

2.1 cm

1.6 cm

1.7 cm

Increased
by 0.1 cm
Increased
by 0.4 cm
Increased
by 0.1 cm

Dark
yellow
brown
color
Dark
yellow
brown
color
Dark
yellow
brown
color
Very
dark
brown
Very
dark
brown
Very
dark
brown
No color
change
No color
change
No color
change

This table of results simply highlights the measurements obtained which are the margins by
which the muscle fiber contracted. Other factor such as color change were also noticed as part of
our observations. As indicated on the table, in solution A the margin of difference was an
average of about 0.1 cm depicting a dark shade of yellow brown. In solution B which contained
ATP + KCl and MgCl2, the margin by which the muscle contracted was very significant. To be
exact it shrinked by an average of about 0.33 cm and the change in color was a more intense
shade of brown. In solution C, no contraction occurred as the results indicate. On the contrary the
length of the muscle fiber increased instead by a margin average of about 0.2 cm and no color
change observed. Based on our data results we can determine that the most effective of all the 3
solutions was solution B.

Discussion:
This muscle contraction experiment was performed in order to observe the effects of ATP
(Adenosine Triphosphate) on a muscle. The muscle in question was the psoas muscle of a rabbit.
In order to be properly analyzed, this muscle fiber was sliced up and placed under a microscope
and its measurement were taken to accurately determine the differential margins after adding
each solution to it. We hypothesized that the fiber would either be affected in the presence of
those solutions by becoming elongated or contracted. The contrast magnification of the
microscope was determined by the length of each strand because they were not all exactly the
same lengths. Some were longer than others. So we used the 10X or 40X accordingly based on
the lengths of each muscle fiber. As previously mentioned solution A contained ATP by itself
mixed with distilled water and we observed a very mild margin difference from its initial

measurement. In solution B where ATP was again present but this time around contained MgCl2
and KCl we observed a significant change in margin averaging exactly about 0.33 cm. in
solution C, only MgCl2 and KCl were added and we observed no change whatsoever. This lead
us to confirm what we had hypothesized from the beginning which was the fact that ATP was the
source of energy in the process of contraction and without it, no contraction can occur.
While performing this experiment, there are multiples sources of errors that could have taken
placed such as the glycerin becoming weakened due to contamination by another solution
causing it not be as effective as it should be for example by not removing the calcium from the
muscle fiber or the muscle contracting due to another chemical being present in the glyceryl
instead of ATP which would give us a false analysis. But relying on the result obtained I am
certain that even if there were errors throughout this experiment it was very minimal because the
desired results were obtained.

Sources Utilized:
http://www.nlm.nih.gov/medlineplus/ency/imagepages/19841.htm

http://www.oxfordreference.com.libproxy.csun.edu/view/10.1093/acref/9780198529170.001.000
1/acref-9780198529170-e-18365?rskey=ybnwnQ&result=7

http://meat.tamu.edu/ansc-307-honors/muscle-contraction/

http://www.oxfordreference.com.libproxy.csun.edu/view/10.1093/acref/9780198524038.001.000
1/acref-9780198524038-e-862?rskey=y351nN&result=5

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