Physiological Laboratory Practice Week One:

written by : Maria Dini Admirati (07120110076), Silvestri (07120110083), Devia Irine P (07120110084),
Valda Garcia (07120110085), Exi Indriastuti (07120110094), Jessica Soeryawinata (07120110096), Keziah
Tuerah (07120110103)

Report

Faculty of Medicine
Universitas Pelita Harapan
Karawaci - Tangerang

Content

Cover 1
Content

1. Introduction3
2. Materials and Methods
3. Results

4. Discussion 10
5. References 13

1. Introduction
Each of your skeletal muscles is a separate organ composed of hundreds to thousands
of cells, which are called muscle fibers because of their elongated shapes. Thus, muscle
cell and muscle fiber are two terms for the same structure. Skeletal muscle also contains
connective tissues surrounding muscle fibers and whole muscles, and blood vessels and
nerves. The sarcoplasm of a skeletal muscle fiber contains hundreds to thousands of long,
cylindrical structures termed myofibrils. Each myofibril is about 1 to 2 micrometers in
diameter and extends the length of the entire muscle fiber. During contraction, the
myofibrils shorten as their component proteins change position. Because myofibrils are
attached to the ends of the muscle fiber, the shortening of the myofibrils during a
contraction causes the fiber to shorten.
Myofibrils consist of bundles of short myofilaments . Whereas a single myofibril runs the
length of the muscle fiber, it takes many successive groupings of myofilaments to run the
entire length of a myofibril.
Thin and Thick Filaments The bundles of myofilaments are classified as thin
myofilaments (usually simply called thin filaments) and thick myofilaments (usually
called thick filaments). Thin filaments are only about 5 to 6 nanometers in diameter. They
are primarily composed of two strands of the protein actin twisted around each other to
form a helical shape. In each helical strand of actin, many small, spherical molecules are
connected to form a long filament resembling a string of beads. Each
spherical molecule is called G (globular) actin, and each filament composed of a strand
of G-actin molecules is called F (filamentous) actin. Two regulatory proteins,
tropomyosin and troponin, are part of the thin filaments. The tropomyosin molecule is a
short, thin, twisted filament that covers small sections of the actin strands. The troponin
has three functions: Structurally, it (1) attaches to actin to anchor itself in place, and (2)
attaches to tropomyosin to hold it in place over the surface of the actin; (3) functionally,
troponin provides a binding site for calcium ions. In contrast, thick filaments are about
twice as large as thin filaments, with a diameter of about 11 nanometers. Thick filaments
are assembled from bundles of the protein myosin. Each myosin molecule in a thick
filament consists of two strands; each strand has a free, globular head and an attached,
elongated tail. The myosin molecules are oriented on either end of the thick filament so
that the long tails point toward the center of the filament and the heads point toward the
edges of the filament and project outward toward the surrounding thin filaments. During
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a contraction, myosin heads form crossbridges by binding thick filaments to actin in the
thin filaments. As mentioned previously, skeletal muscle has striations. This striated
appearance is due to size and density differences between thick filaments and thin
filaments. Under the light microscope, two differently shaded bands are observed. The
dark bands, called A bands, contain the entire thick filament. At either end of a thick
filament is a light band region occupied by thin filaments that extend into the A band
between the stacked thick filaments. The light bands, called I bands, contain thin
filaments but no thick filaments. In addition to the thin filaments in I bands, there are
protein filaments called titin, which play a role in muscle elasticity, control of thick
filament assembly, and passive stiffness generated in the muscle. Within both the A bands
and the I bands, other important structures can be clearly observed using an electron
microscope:
The H zone is a light, central region in the A band. It is lighter shaded because only
thick filaments are presentthat is, there are no thin filaments overlapping the thick
filaments in the H zone in a relaxed muscle fiber. At maximal contraction, the thin
filaments are pulled into this zone, and the H zone disappears.
The M line is a thin transverse protein meshwork structure in the center of the H zone
of a relaxed fiber. It serves as an attachment site for the thick filaments and keeps the
thick filaments aligned during contraction and relaxation.
The Z disc is a thin transverse protein structure in the center of the I band that serves as
an attachment site for thin filament ends. Although the Z disc is circular, when viewed
head-on, only the edge of the circle is visible, so it sometimes looks like a line.
Connectins are Z disc proteins that anchor and interconnect the thin filament ends at
either end of a sarcomere.
The events of muscle contraction have been called excitationcontraction coupling,
meaning that the stimulation of a muscle fiber by a nerve impulse results in a series of
events that culminates in muscle fiber contraction. First a nerve impulse causes ACh
release at a neuromuscular junction. ACh binds receptors on the motor end plate,
initiating a muscle impulse. Second The muscle impulse spreads quickly along the
sarcolemma and into the muscle fiber along T-tubule membranes, causing calcium ions to
be released into the sarcoplasm. Third Calcium ions bind to troponin, causing
tropomyosin to move and expose active sites on actin. Myosin heads attach to the actin
and form crossbridges. Fourth Myosin heads go through cyclic attach, pivot, detach,
return events as the thin filaments are pulled past the thick filaments. ATP is required to
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detach the myosin heads and complete the sequence of cyclic events. The sarcomere
shortens, and the muscle contracts. The cyclic events continue as long as calcium ions
remain bound to the troponin. Fifth Calcium ions are moved back into the sarcoplasmic
reticulum by ATP-driven ion pumps to reduce calcium concentration in the sarcoplasm,
leading to relaxation. Termination of the muscle impulse results in the passive sliding of
myofilaments back to their original state.
Muscle fibres are multinucleated cells responsible for muscle contractions. Each
muscle has between 10 000 and 450 000 fibres. There are two main types that can be
distinguished by their colour. Red muscle fibres (also called Type 1 or slow-twitch fibres)
acquire their colour from their good blood supply and high myoglobin content. They
fatigue slowly, and use glycogen and fat as their fuels. White muscle fibres (also called
Type 2 or fast-twitch fibres) have a more moderate blood supply, only use glycogen as
fuel, and fatigue more quickly than the red fibres. However, they are bigger than the red
fibres and their contractions are much more powerful. Red muscle fibres are adapted for
aerobic activities of long duration. White fibres are adapted for short bursts of explosive
anaerobic activity.

2. Materials and Methods

2.1 Materials
-

Ergograph
Sphygmomanometer with
Metro
Stopwatch
Stationary dynamometer

2.2 Methods
2.2.1 Effect of rapid (high frequency) muscle work, rest and massage on muscle
fatigue
1. Choose one student as a subject to perform the first experiment and the
other student as an experimenter.
2. Let the subject sit down and place his/her dominant arms elbow on the
table and positioned the arm and the index finger as shown in the photo,
ready to pull the trigger.
3. With the stopwatch in the experimenter hand, the experimenter gives
him/her a signal to perform finger aerobics exercise (by pulling the
trigger) with the frequency once every second.
4. Let the subject continue pulling the trigger until he/she can no longer do
so (become fatigue). Record the time and the number the finger
movement from the beginning until he/she stop pulling the trigger.
5. Let the subject rest for 1 minute.
6. Repeat steps 3 and 4.
7. Record the new time and number of finger movements.
8. Let the student rest for 1 minute and massage his/her lower arm.
9. Repeat steps 3 and 4.
10. Record the new time and number of finger movements.

2.2.2

Effect of low frequency muscle activity

1. Choose another subject
2. Repeat exp. 2.2.1, but with frequency 1 every 4 seconds.

2.2.3

Effect of changes in vascularization on muscle activity

1. Use the same subject.
2. Set the sphygmomanometer nunchette (cuff) in his/her right upper arms.
3. Let the subject pulls the trigger once every 4 seconds for twelve times.
4. In the 13th inflate the cuff until the right radial pulse disappear. Record
the time at which the pulse disappear.
5. During and within inflation, the subject continuous pulling the trigger
until he/she have no more energy so pull the trigger (become fatigue). At
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that time, deflate the cuff completely, so the blood flows return to
normal. Record the time when the cuff being deflated ( from beginning
until the end of the experiment, the subject continues to pull the trigger).
6. The strength of the pulling power will return to normal.
7. Stop the experiment when the pulling power returns to normal.
2.2.4

Effect of body position on muscle initial length

1. Choose one student as a subject and the other student as experimenter.
2. Let the subject sits upright on the dynamometer facing the balance.
3. Set the hook to her right ankle.
4. The experimenter, by facing the balance, gives signal to the subject to
flex his right lower limb backward as strong as possible, and then record
the digit in the balance that indicate the strongest result of the subject.
5. Repeat the step 3 and 4 three times.
6. Let the subject sits and bends her body forward as far as possible.
7. Repeat step 3 to 5.
8. Let the subject still in his place but now lay his body down backward.
9. Repeat again step 3 to 5.
10. Let the subject moves to the other side of the dynamometer and sits with
the balance in her back.
11. Set the hook on her right ankle.
12. Repeat the step 3 to 9.
13. Record the result of the experiment in a table.
14. If you have time, you can repeat for the left limb of the subject.

3. Results
EXPERIMENT MUSCLE FATIGUE
EXPERIMENT 1
No.
1.
2.
3.
Question:

Time
60s
60s
52s

Number of Finger Movement

43 times
42 times
44 times

Speed
0,716667
0,7
0,846154

1) Yes.
EXPERIMENT 2
4 minutes 26 seconds 70 times

EXPERIMENT 3
No.
1.
2.
Note:

Time
2 minutes
30 seconds

Number of Finger Movement

20 times
5 times

1. Data being recorded from the first time the cuff is being inflated and the pulse
disappeared until the subject feels fatigue.
2. Data being recorded from the time subject feels fatigue and cuff is being deflated
until the pulse returns.
EXPERIMENT EFFECTS OF BODY POSITION ON MUSCLE INTIAL LENGTH
EXPERIMENT 1
Sit
26 kg
34 kg
36 kg

Facing the Balance

Sit and bend
Lie Back
14 kg
20 kg
17 kg
23 kg
17 kg
22 kg

Facing Away from the Balance

Sit
Sit and bend
Lie Back
12 kg
13 kg
13 kg
11 kg
13 kg
15 kg
10 kg
11 kg
15 kg

4. Discussion
1. Does muscle fatigue happen? What is the effect of rest and massage?
The inability of a muscle to maintain force of contraction after prolonged
activity is called muscle fatigue. Fatigue results mainly from changes within muscle
fibers. Even before actual muscle fatigue occurs, a person may have feelings of
tiredness and the desire to cease activity; this response, called central fatigue, is
caused by changes in the central nervous system (brain and spinal cord). Although its
exact mechanism is unknown, it may be a protective mechanism to stop a person
from exercising before muscles become damaged. As you will see, certain types of
skeletal muscle fibers fatigue more quickly than others. Although the precise
mechanisms that cause muscle fatigue are still not clear, several factors are thought
to contribute. One is inadequate release of calcium ions from the SR, resulting in a
decline of Ca2_ concentration in the sarcoplasm. Depletion of creatine phosphate
also is associated with fatigue, but surprisingly,the ATP levels in fatigued muscle
often are not much lower than those in resting muscle. Other factors that contribute
to muscle fatigue include insufficient oxygen, depletion of glycogen and other
nutrients, buildup of lactic acid and ADP, and failure of action potentials in the motor
neuron to release enough acetylcholine.
Localized muscle fatigue is defined as the fatigue that is localized to the
muscle or group of synergistic muscles performing contraction. Localized muscle
fatigue can be induced by sustained muscular contractions and is associated with
such external manifestations as inability to maintain a desired force output, muscular
tremor, and localized pain. In a clinical and sports setting, massage is utilized widely
and believed to be efficacious in the recovery from muscular fatigue. Previous
studies that investigated the effect of massage on muscle fatigue evaluated the loss of
force generating capacity by measuring the rate of decline based on maximal
voluntary contraction. However, it has been indicated that the results of muscle
fatigue assessed by this method tend to be influenced by subjective qualities, such as
personal motivation and current level of pain. More objective and reliable measures
are necessary to evaluate the possible efficacy of massage against muscle fatigue.

2. Does muscle fatigue happen?

Muscle blood flow has been characterized as being pulsatile during rhythmic
exercise (6), such that the mean blood flow is a summation of the flows that occur
during the contraction and noncontraction periods (see Fig. 1). When the vasculature
of the muscle is forcefully compressed during the contractile phase of the duty cycle,
arterial inflow is inhibited and blood is rapidly expelled from the muscle into the
extramuscular venous circulation. On muscle relaxation, arterial flow into the muscle
is restored and venous flow returns to steady-state values. At the onset of steadystate contractions, before neural, metabolic, endothelial, and myogenic factors are
capable of initiating substantial vasodilation, the initial contractile expulsion of
blood from the muscle has been suggested to contribute significantly to the
immediate increase in muscle blood flow . However, during steady-state rhythmic
contractions, after the vasodilatory processes have been successfully activated, it is
controversial whether the contractile ejection of blood flow (muscle pump effect)
is a determinant of steady-state muscle flow .
Irrespective of whether the muscle pump effect is a significant determinant
of the overall blood flow response during steady-state rhythmic contractions, it is
clear that the contraction and relaxation phases of a single contractile cycle result in
very different temporal blood flow profiles during steady-state contractions , with
each phase contributing quite differently to the overall steady-state blood flow
response. It would be expected that the contribution of the contractile phase to
overall mean blood flow during rhythmic contractions may depend on the strength of
the contraction, the volume of blood contained in the muscle at the onset of
contraction, and the ratio of the time in which the muscle is in the contractile and
noncontractile phases (duty cycle).
As has been demonstrated,arterial blood inflow into contracting muscle is
significantly diminished during the contractile phase owing to vascular compression,
and muscle perfusion is reestablished during the noncontractile phase. If the duration
of the contractile phase of the duty cycle is large, then this can have detrimental
effects on overall muscle perfusion and result in inadequate delivery of O2 and
diminished muscle contractility (fatigue). In addition, if the frequenci (and not
necessarily duration) of contractions becomes so great that blood flow and filling
between contractions is of a short duration, then the mean muscle blood flow can
again be compromised and contractility may be attenuated . In the present study, the
frequency and duration of contractions was below the threshold for any negative
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effect of this impediment of blood flow to occur, and muscle contractility was not
affected, as demonstrated by the maintenance of steady-state force development at
all contraction frequencies.
An important component of the present study was that muscle work and O2
uptake rose proportionally with contraction frequency. Therefore, the changes
demonstrated in the contribution of the contractile and noncontractile phases to
muscle blood flow were influenced by both the increased metabolic rate and duty
cycle. Mean venous muscle blood flow rose proportionally with increased
contraction frequency, demonstrating the tight coupling of O2 supply to increased
metabolic demand. The factors that are known to couple muscle blood flow to the
metabolic rate during steady-state rhythmic contractions are numerous and complex
in their interaction.

5. References
McKinley,Michael. Human Anatomy 3rd edition. McGraw Hill :
United States.2012
Tortora, Gerard C. Principles of Anatomy and Physiology 12th
edition: United State. 2009
http://jap.physiology.org/content/95/3/1139.full

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http://www.answers.com/topic/muscle-fibretypes#ixzz23CKSERSC

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