PROPERTIES OF CARDIAC MUSCLE

Aim
The aim of this chapter is to study the various mechanical properties of
cardiac muscle.

Introduction
In the laboratory, the preparation that is usually used for this experiment is
the frog heart. Although structurally the frog heart is very different from that
of the mammalian heart, in terms of function, the frog heart and the
mammalian heart have very similar properties.

The anatomy of the frog heart is shown in Fig. 7A. Unlike the mammalian
heart, the frog heart is made up of only 3 chambers - two atria and one
ventricle. Deoxygenated blood enters the right atrium from the sinus
venosus, while the left atrium receives oxygenated blood from the pulmonary
vein. Although the ventricle is not divided into two by a septum, folds found
within the ventricle tend to separate the deoxygenated blood from the
oxygenated blood, so that the blood can be channelled into the pulmonary
artery and aorta, respectively.

Similar to the mammalian heart, the frog heart also beats sPontaneously and
an
rhythmically. The heartbeat of the frog d other cold-blooded animals arises
from the sinus venosus, a structure that is equivalent to the sinoatrial node
found in mammalian species. In other words, the cardiac pacemaker of the
frog heart is located in the sinus venosus. Similar to the mammalian heart,
the number of impulses that arises from the pacemaker in the sinus venosus
is influenced by sympathetic and parasympathetic nerves, temperature,
chemical agents and other factors.

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Procedure
This experiment can be performed in groups of 4 to 8 students. The spinal
cord of the frog is first pithed and destroyed. The frog is subsequently laid out
prone on a wax tray and the knee pinned to the wax. The thorax is cut to
expose the heart. The connective tissue is teased away and the heart
isolated. The isolated heart is kept moist with Ringer solution to prevent
drying.

A. Normal Heart Rate

Using a clip and a string as shown in Fig. 7B, the isolated heart is
attached to a force transducer, which is connected to a recording
device. Obtain an adequate strip of recording of the normal contraction
of the heart and label it as "Normal heart beat". Identify each
component of the heart beat on the recording.

B. Effect of Vagus Nerve Stimulation

In the frog, because of the difficulty in isolating the vagus nerve to the
heart, direct stimulation of the vagus nerve is not possible. Instead, the
junction between the sinus venosus and the right atrium is stimulated
for studying the effects of vagus nerve stimulation. This site, known as
the crescent of the heart, is where the vagus nerve enters the heart.
An electrode connected to a power source can be used to stimulate
this area.

The stimulation is set for a frequency of 20 pulses/sec for a duration of

10 ms, with a strength of stimulation of 10V and the number of
impulses at 60. Carefully place the electrode on top of the crescent of
the heart as shown in Fig. 7B and begin recording the normal heart
beat. After a few normal beats have been recorded, stimulate the
vagus nerve and observe the effects on the heart rate and strength of
contraction of the heart. Should there be no response from the

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 stimulation, adjust any of the parameters of strength, duration or
frequency, and repeat the stimulation.

Usually, contraction of the heart becomes chaotic during stimulation.

When the stimulation stops, the heart may also stop beating for a few
seconds. This is because the effect of the stimulation continues for
several more seconds after the stimulation has stopped. Immediately
afterwards, the heart beat will return to normal.

C. Effect of Temperature
Prepare a beaker each of cold (10°C, chilled in ice) and warm (40°C,
heated) Ringer solution. Obtain a few normal heart beats. Then, using
the Pasteur pipette, add cold Ringer solution on to the heart drop by
drop. Label the recording "Effect of cold Ringer" and observe the effect
of cold Ringer solution on the heart rate and amplitude of contraction.

Wash the heart with normal Ringer solution several times and allow the
heart rate to return to normal.

Repeat the same procedure with the warm Ringer solution .

D. Effects of Noradrenaline, Adrenaline and

Acetylcholine
Record several normal heart beats and then add a few drops of
noradrenaline solution (0.1 mg/ml) on to the heart. Label the recording
"Effect of noradrenaline" and observe the effect of noradrenaline on
the heart rate and amplitude of contraction. Wash the heart with
normal Ringer solution several times and allow the heart rate to return
to normal.

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Repeat the same procedure with adrenaline (0.1 mg/ml) and
acetylcholine (1 mg/ml).

E. Heart Block (Stannius Ligature)

In the mammalian heart, under normal conditions, the SA node
determines the frequency of the heart beat as the rate of impulses
generated by this structure is the highest compared to other parts of
the heart. Should the SA node or the pathway leading from it is
damaged, the AV node with the next highest rate of discharge would
take over and become the pacemaker of the heart.

The hierarchy of the conduction system of the heart is such that the
node or tissue that discharges with the highest rate becomes the
pacemaker. This can be demonstrated by Stannius Ligatures (Fig. 7C).

Bathe the heart with Ringer solution and allow the heart rate to return
to normal. Using a pair of fine forceps, pass a piece of thread through
the space between the sinus venosus and right atrium (Fig. 7C), draw
the ends of the thread together and then make a loose knot (First
Stannius Ligature).

Then, pass another piece of thread between the atria and the ventricle
and make another loose knot (Second Stannius Ligature).

Obtain a recording of the normal heart rate. During the I recording,

tighten the First Stannius Ligature so as to isolate the sinus venosus
from the right atrium. This will cause the ventricle to stop contracting.
Observe whether the atrium continues to contract and whether the
ventricle resumes its contraction. Observe the frequency of the heart
beat.

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 Next, tighten the Second Stannius Ligature so that the atria are
isolated from the ventricle and observe the effect on the heart rate.
Compare the sequence of contraction between the sinus venosus, right
atrium and ventricle after the Second Stannius Ligature.

Fig-7 (A) Chambers of the frog heart

(B) Isolated frog heart connected to a transducer

(C) Stannius Ligatures I and II