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 ********************

CHAPTER m2.12
Recording of a Normal Cardiogram of
Frog's Heart and Effect of
Temperature on it

Atrial systole

To record a normal cardiogram of frog's heart and the effect Atrial diastole

MM
f temperature on it.

HEORY

heart consists of two atria and one ventricle.

The frog's
Sinus venosus is the pacemaker of frog's heart and the
the atria and then
umpulse travels from sinus venosus to
the
the ventricle. Electrical events in the heart precede
mechanical events
mechanical contractions. The different
recorded atrial systole, atrial diastole and ventricular
are
Systole followed by ventricular diastole. The recording Ventricular diastole
entricular systole
obtained by the various mechanical events of the frogs

heart is known cardiogram. of frog's heart.

as
Fig. 2.12.1: Normal cardiogram

BORMAL CARDIOGRAM 7. Time/signal marker

e Cardiogram is a record of the mechanical activity or tne 8. Dropper, thread, cotton wool and pins.

eart, while electrocardiogram is a record of the electricial Speed of the drum: 2.5 mm/sec
y of the heart. In this and the following experiments,
emechanical events of the heart will be recorded wtn & PROCEDURE

Narling's heart lever (Fig. 2.12.1).

Exposure of the Frog's Heart
APPARAT 1. Stun and pith a frog and lay it on its back inmidline
a dissection
from
incise the skin in the
1. Myograr board
tray. Usinga scissors, end of this cut
2. Lucas crAnber
Xiphisternum to the jaw. Extend the lower
anterior
. both pieces of skin. The
Disseci on apparatus laterally and remove

4. A chest wall is now exposed.

pan Ringer's solution (cold and warm) horizontal cut in the muscles at the levei of

Aymogr ph with drum 2. Give a

the abdominal wal,
xiphisternum (do through
Starling's heart lever not cut
.
 otherwise the viscera will
and
spill out ) Using bone forceps the heart and pass the sharp nook ot the
sei**os. cut
through
emove t h e chest wall in
the
pectoral girdles and the
bent pin thr.
apex ol the ventricie, taking care not
be
revealed beating in its
one
piece. The heart will now cavity.
to punesgh
e ts
the pericardial
pericardium and remove it sac. sit througn Lift the heart gently by raising the lever and
the right up to the base
ot position, adius
heart. Removal of pericardium is important SO that ts movements are
satisfactor
help to
support the heart tirmly from the apeN And
as it
will mean position is horizontal. andin
variables which are
used will come in direct
contact with
une NOTE
the heart muscle.
During systole, the lever is pulled down, whie
By inserting a during diastole th
pin
of the heart to the
through bulbous (base). fix the base
the Spring or the lever puiled up to its former position.

myographboard so that the

heart is not
pulled from both the ends and thecontracting Move the stand carrying the
preparation
iS not affected. recording so that the lever at
and the leve
Examine the heart
1s a
tangent to
tne cylinder and the
carefully (Fig. 2.12.2). There is Writing point is lightly touching the cylinder sur.e
one ventricle, separated from
the two atria by the Record the cardiac activity 1or
atrioventricular groove and bulbus with the drum moving at a
about 15 cm on the paper.
arises trom the
arteriosus which
ventricle and divides into two aortae. 4. Take thhe time tracing
speed ot 2.5 mmis.
5. Lift
5
every seconds with the help of
the ventricle
up and find behind it
the sinus venosus
time signal marker below
the graph obtained.
with the two venae cavae 5. Note
emptying into it. the number ot
peaks recorded and
A carerul observation will reveal
calculate the
frog's heart rate.
the white crescentic
line between the sinus venosus and right atrium. This
1S the site ot the
RemakS NOTE
and
Bidder 's ganglia of the Normal heart rate of frog varies from 30/minute to 50/minute.
Vagus nerves.
The color of the
ventricle becomes pale during systote Effect of Temperature
as blood is forced of it. Feel the
out
hardening of the on Frog's Cardiogram (Fig. 2.12.3)
ventricle when it contracts. There valves in the
are no . Pour
amphibian Ringer's solution at room
temperature
trog's heart. on the heart fixed in the Lucas
chamber; note the
Sequence of heartbeats: The contractions of the temperature of the Ringer's solution. With the dum
four units of the heart are
progressive. The sinus running at slow speed (2.5 mm/sec) pour Ringer's
leads off, followed by the atria, ventricle and bulbus solution
in that order. The rate ot the heart
warmed to about 40°C on the heart, until there
depends on the 1S an
obvi1ous jncrease in its rateand force.
frequency of the sinus, as it is the pacemaker. 2. Stop kymograph and pour Ringer's solution at room
the
temperature on the heart till it resumes the previous rale
Recording of Normal Cardiogram and torce.
. Transfer the frog to the Lucas chamber. Fit the Starling's n e n g a n record the effect of cold Ringer's solution at
heart lever the vertical rod of the stand about 10C in the same
on
directly above manner.
4. lake the time tracing
every 5 seconds with the heip o
time signal marker below the
obtained. Label graph your
Hook cotton
thread
tracing. indicating with arrows, the points where
hotan
cold Hinger's solution was applied. Calculate the heart

Ventricle
rate at these temperatures and enter the data in your
workbooK.

Crescentic ine
NOTEAt
Truncus high temperature, there is an increase in u e
arteriosus activity of the pacemaker celis wwhich
ardia
h i c h g e n e r a t e s more
cardiac
mpulses per unit time .
ge rt rate and
"creasing the heart rate and
Left aortic arch Sinus venosus
Common neight of contraction activity of
the
etabolic
Opening o inferic
ng
cells of the atria an à n d decrease
in the
the thorax vena Cava increase the force of e
s1y
old has the opposite effects due to decrease in the metao
Left atrium
activity.
Fig.2.12.2: Frog's heart
 hapter 2.12 Recording of a Normal Cardiogram of frogs Heart and Efect of Temperature on it 47

Atia tystole
Atr diaetoie

entmcie [Link]
Cotd (10
Warm (40 C) HR 2m
24imin HR 36/min Time marker5/sec)
Nomal (25 C)-HR

Fig.2.12.3: Effect of temperature on cardiogram.

diastole, then ventricular systole (this is the strongest of the

PRECAUTIONS four contracting units) is followed by diastole (contraction
of sinus may also be recorded). The heart rate is beats/min
Care shouid be taken not to damage the heart during
dissection.
and the
rhythm is regular).
Q3. What is the mechanism of action of hot and cold Ringers
Pericardium should be removed before fixing the heart
solution on frog's heart?
on the myograph board practical? of this
. Record the normal cardiogram before recording the Q4. What i5 the physioclinical significance
Q.5. Why itis important to fix the base of the heart?
effect of warm and coid Ringer's solution. Q6. Why it is necessary to remove the pericardium?
The temperature of warm Ringer's solution should not Q7. What ill happen if only the ventricle is warmed?
be more than 42"C. Warming of ventricles increases only the height of the
. Effect of warm Ringer's solution should be recorded contraction but the rhythm remains unchanged.
before the effect of cold Ringer's solution.
Do not puncture the ventricle while passing the pin 0BSERVATION AND RESULT
through its apex.

Recording ofeffect of temperature on frog's cardiogram.

PHYSIOCLINICAL SIGNIFICANCE Heart rate (beats/ Height of
min) contraction (cm)
This practical shows that heart rate changes in the human
ay with change in internal body temperature, e.g. NOmal
ercise, fever and hypothermia.
Warm Ringers
QUESTIONS solutlon (

. Why is frog's heart used for the study of properties of ners

30lution { . . )
heart?
ne frog heart continues to beat even after the chest is
opened. It obtains an adequate supply of oxygen directly
t e Dlood in its chambers and from the atmosphere NTERPRETATION
has no coronary circulation). Its rate is sufliclently slow
o allow observation of its sequence. Also, it continues to
Tunction over a wide range of temperature. The properties
s s * n e t e s e e * e e t u a a s s s d s ssepese*teassseweesessss******************

which can be studied include

excitability, automaticity
nyhmicity, contractility, conductivity, retractoriness ana
all-or-none law.
"***ssasesessstaSas **e********************* ***tenaess********

4. Describe the graph obteined by you. ***

he
downstroke of the tracing represents systole and-the
upstroke diastole. There is atrial
systole followed by atrial
 Section 2: Amphibian (Frog) Experiments

STUDY NOTESs

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 CHAPTERR2.13
Date*ssneenenee***********

Properties of Cardiac Muscle

AIM
is because the next normal impulse coming from the
SA node reaches in the absolute refractory period of
To study the properties of frog's heart. extrasystole and hence fails to evoke a response. The
response which follows the compensatory pause is of
THEORY greater magnitude than the previous o n e because of the

The following properties will be studied in a beating

accumulation of calcium ions during the pause
3. Refractory period: The cardiac muscle has a long8

heart: refractory period. This is because the action potential

a. Autorhythmicity of cardiac muscle has a long duration of about 300 ms.
b. Extrasystole and compensatory pause During much of the action potential (about 250 ms), the
c. Refractory period. cell is completely refractory to further stimulation, i.e. it
2. The following properties wiil be studied in a quiescent
it stimulated.
is unable to fire no matter how strongly is
heart (after Stannius ligatures):
a. Summation of subminimal stimuli.
This unresponsive state is calledas absolute
refractory
period. During the latter part of action potential (about
b. All-or-none law 50 ms), the cell is able to fire a second action potential
Staircase phenomenon. provided a stronger than a normal stimulus is given.
This period is called as relative retractory period.
APPARATUS The cardiac muscle action potential duration is almost
equal to the duration of mechanical activity. Therefore,
1. Same as in recording of normal cardiogram (Refer
the mechanical responses of cardiac muscle cannot be
Chapter 2.12)
2. Induction coil summated or tetanized.
3. Wire electrodes and signal marker.
Speed of the drum: 2.5 mm/sec (Slow) PROCEDURE
1. Set up the experiment as for recording a normal
PROPERTIES INABEATING HEART cardiogram.
Extrasystole: When the heart is stimulated [by an 2. Apply electrodes on the ventricle of heart.
puise oher than that originates from the sinoatrial
(SA) nodel during late diastole (relative refractory
3. The distance between primary and secondary coil is so
adjusted that a minimal stimulus is obtained.
eOd, the heart
comes earlier than
muscle may contract. This contraction 4. Use the electromagnetic signal marker to indicate the
the normally expected contraction. application of stimulus.
This is called
extrasystole. 5. A few normal beats are recorded.
mpensatory pause: The pause (silence) following 6. A "break" shockis applied by opening the short circuiting
ne
extrasystole is known as
compensatory pause. This key and thereafter the primary key.
150 Section 2: Amphibian (Frog) Experiments

Simulate the ventrH le duing the ditterent phases ol period. More than one subminiinal stimuli add
dd ugethr
the cardiac
civle, 1 e
sstole. carh and late diastole and and may take the membrane ntial
potential to the fi
8
Tecord the etfects so as to generate an action potential iring levw
The graph labeled properly show the systole
is
to and
diastole the cardiogram. application of extra
in

point of PROCEDURE
stimulti extrasystole and compensatory pause. A ume
tracing IS also
taken. Effect of Stannius Ligature
NOTE . Set up the experiment as for
recording
orma
When the a nore
stimulus falls during any part of systole, it has no effect cardiogram
gs2131A to
El, the heart continues
to beat as before. But when
the stimulus talls . Pass igature thread under the truncus
a
during diastole. the heart contracts
immediately. arterins
and the atria, bring its ends to the dorsum of
contraction is calied extrasystole or premature beat,
This extra
the heam
s
followed by Ensure that the ligature thread is placed at the
pause called
"compensatory junction
a

normal contraction, the extrasystole pause". Duration

and compensatory pause
or of sinus venostus and atria (on the white
is
equal to twO normal cardiac cycies crescentie
line). This is First Stannius ligature.
3. Record some normal beats, Stop
Ihis experiment also shows the tne arum, then tie
the thread with a single knot over the white
properties ot
crescentic
excitability, contractility, autorhythmicity and line.
conductivity (stimulation of any part causes contraction 4. fit has been
of the rest of the appied properly, the sinus wil
continue
heart). to beat asbefore, while the atria and ventricle
willstop
during diastole after one or two beats (Fig.
2.13.1B).
PROPERTIES IN A QUIESCENT HEART (AFTER They will begin to beat after a variable period of 10-
STANNIUS LIGATURES) 20 minutes at a slower rate. This is atrial
Record this activity.
rhythm.
1. Effect of Stannius ligature: Stop the drum and tie the second

First Stannius ligature: Whena

Stannius ligature.
ligature is applied over 5. ond Stannius ligature: Tie a
tight ligature between
the sinoauricular
junction and tied, the conduction the atria and ventricle, the ventricle
ot
impulse from sinus venosus to atria is
blocked. immediately
Therefore, atria and ventricle stop stops contracting,
while the sinus and atria continue
sinus venOsus continues to beat. After
[Link] the to beat at their own
different rates (the first Stannius
sometime, the ligature is still in place). The drum is restarted and
atria start to generate their own impulses at a slower the recording is taken
rate and the atria and ventricle start contracting. This when the ventricle has stopped
is atrial rhythm. beating.
6. The drum is
Second Stannius ligature: When ligature is applied a
again stopped. Wait for some time till the
ventricle starts
between the atria and ventricle and tied, the contracting. This is idioventricular
transmission of cardiac impulse is blocked from the
rhythm. And the contractions are recorded.
7. The time
atria to the ventricle. The ventricle
stops beating
tracing is recorded. The rate of the
whereas the atria continue to beat. After sometime,
three
rhythms-sinus rhythm, atrial rhythm and
idioventricular rhythm is calculated
the ventricle starts
generating its own impulses and 8. Summation of Subminimal Stimuli:
(Fig. 2.13.1B
Starts beating at a rate of 15-40 beats/min. This is
called idioventricular rhythm.
a.
The primary and secondary coils are kept at
tne
farthest distance
and then they are
2.
All-or-none law: It is applicable to the whole heart as it
brought closer till the break shock is graduauy
acts as a functional syncytium because of the presence produced.
ol gap junctions. Therefore, with threshold stimulus
D.
The secondary coil is now moved a little away from
neart will contract to its the
maximum and it will not respond
the
primary coil so as to find a stimulus that
t the
stimulus is below the tails to produce contraction
a
threshold. of the heart.
Staircase C.
APply this stimulus several times at the interval
phenomenon: It refers to the successve
increase in the first 0.5-1 second till the o
few cardiac ventricle shows
the silent heart has contractions atter The 1Oth-20th contracu
stimulus
the beneficial
started to beat. It is
because o contraction. Each
generally gives a u
effect of the subminimal stimulus produces
(accumulation of calcium ions). previous conitaction some changes in the heart muscle,
Summation of subminimal more excitable to the
successive stimulus.
makingnce
stimulus generates a stimuli: A subminimal the
graded potential which summation of potential
subminimal stimuli reacheschanges
summated because it does not have can be induced Dy
to threshold,
any refractory ventricle contracts as shown in u
Figure [Link].
 ompensatory pause Compensatory pause

WTN111MVVVVUV
Treppe

Extrasystole Extrasystole
-Signal marker
Earty diastole Late diastole
AExtrasystole (ES) and compensatory pause (CP)

Normal Atrial rhythm Ventricular thythm

HR 36 bpm ,1st ligature 24 bpm 2nd ligature Dpm

Time marker

Heart biock: Stannius ligatures

Threshold Maximal Supramaximal

Subthreshold

Signal marker

All-or-none taw

Signai marker

Dalrcase phenomenon (beneficial effect

e r

Summation ofsubminimal stlmull

Figs.2.13.1Ato E: (A) Extrasystole and compensatory pause; (B) Effect of Stannius ligatures; (C) All-or-none la
(D) Stalrcase phenomenon (beneficial effect; (E) Summation of subminimal stimuli.
(HR: heart rate; bpm: beats per minute)
 All-or-none Law: QUESTIONS
Starting with subthreshold stimuli. gradualy
ncrease the streigth of stimuli until there is a Q.1. What Is the catuse or c o m p e n s a t o r pause afte
after an
extrasystole?
contraction (threshold response). During the extrasystole, when the usual cardiac
b Continue to appli single stimulus of
increasing from the pacemaker reaches the ventricle, it finds that

strength. alowing
between each response
at least 30 second intervals
already contracting (due to gxtrasystole) and in the absolute
refractory period (ARP), SO t has no eftect. The ventricle h
C. Note that though the strength of each stimulus is theretore, to wait tor the next impuse from the sinus to affive
successively increased but the force of contraction before it can c o n t r a c t - n e n c e the briet pause.

remains the same.

10. Staircase Effect
C.2. VWha i licpotentiation"?
a. Three to four threshold stimuli are given one alter
The first contraction atter the compensatory pause is
the other at an interval of 2-3 seconds, so that a new
often more forceful than usuai heartbeat. This is due to
the
increased availability ot intracelular calcium.

contraction occurs immediately after the previous 3. Do ei n humans? What is the

normal
relaxation is over. requenc oie?
D. Note the sutcessive increase in the force of Q4. How will yitraTt1ate between atrial and ventricular
contraction of the ventricle due to beneficial effect. extrasystule?

Atrial and ventricular extrasystole can be distinguished with

PRECAUTIONS the help of an electrocardiogram (ECG).
Q.5. What is the cause ot the long refractory period of the
1Jo study the phenomena ot extrasystole and compensa- cardiac muscle? Whdt is its advantage?
Q6. What is all-or-none law? Which
tory pause, the stimulus has to be applied during late obey this law?
Are all-or-none law and the Starling's Taw of the heart
tissues
diastole.
2. To study the staircase phenomenon, the ventricle should incompatible?
For the all-or-none law to be
be stimulated repeatedly after every 2 seconds. applicable, all the experimental
conditions are to be kept constant. This does not
3.
For studying the summation of etects due to
repeated in
Starlings law of the heart where the initial length of cardiac
happen
subminimal stimuli, the stimuli (subthreshold) should muscle changes resulting in increased force of
be applied repeatedly after 0.5-I seconds. contraction.
Q.7. What is staircase
phenomenon and what is its physiological
basis?
PHYSIOCLINICAL SIGNIFICANCE Q.8. To demonstrate the all-or-none
law phenomenon, why
should the interval between
Extrasystole: Extrasystoles are commonly seen in medical the stimuli should be 30
Seconds?
practice. Ihe common causes are: Ihis will prevent summation of stimuli or beneficial effect.
1. Physiological causes Q.9. With the help of a
a. Smoking diagram describe the properties of tne
cardiac muscle of a
b. Excessive intake of tea
frog?
or coffee
C. Anxiety
d. Lack of
OBSERVATION AND RESULT
sleep.
2. Pathological causes
a. Hyperthyroidism *************************************************************** ****

b. Hypoxia *******************************
********************************** ****

C. Electrolytes imbalance
d. Myocardial damage ****************************************************************************** ******

e.

Some
Digitalis overdose, etc. ***************************************************************************
ectopic foci (from other than
in the atria or the
ventricles generate an
the normal site) ******************************************msssaas** usass *********
********************uos***********

impulse, which
causes an
extrasystole (premature beat). INTERPRETATION
About 2-4 extrasystole/minute are
in humans. considered iomal
******************************************************************************
If the
irequency of extrasy'stoles is >6/min, that **** ***

may indicate some pathological condition.

*****

Heart block: Whenever there ****************************s*s

is *************ssnsmme***********
a
stoppage of
impulse
transmission
Three
from atria
to ventricle, it is called heart block.
degrees of heart biock can occur in humans (1. Ii and
*********************************************************** **************
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 STUDY NOTES

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Date. ***********************

Effect of Stimulation of Vagosympathetic

CHAPTER2.14
Trunk and White Crescentic Line on
Frog's Cardiogram

the skull to the hyoid bone, it crosses a

AIM as
very shiny
tendon.
To study the effect of stimulation of vagosymapthetic trunk b. Lift up the lower border of the muscle and to find the
and white crescentic line (WCL) on frog's cardiogram. vagosympathetic trunk and carotid vessels crossing
the shiny tendon.
THEORY C.
Expose the other trunk also. Put loose ligatures around
them, so that they can be lifted up for stimulation.
The vagosympathetictrunk contains preganglionic para-
sympathetic nerve fibers and postganglionic sympathetic
d. Include the Neef's hammer (for
repeated stimuli) and
fibers (in mammals, the two signal marker in the primary circuit.
systems sepatdeyu
run
2. Stimulation of vagosympathetic
trunk: Record a tew
there are
many more vagal fibers than sympathetic. There-
normal beats then stimulate the
tore, vagal effects usually predominate. for 4-5 seconds with the
vagosympathetic trunk
The stimulation of vagal fibers decreases heart rate and help of Neef's Hammer. Note
the stoppage of the heart
conduction, but it has no effect on ventricular contraction. during diastole. This is vagal
inhibition (Figs. 2.14.1).
The
white crescenticline is formed by the postganglionic
parasympathetic neurons located at the junction between dimuaton ot WCL: After normal beats are restored,
sinus venosus and atria. Stimulation of WCL will have the stimulate the \WCL. for a few seconds and note cardiac
same effect the stimulation of inhibition as above. Repeat steps 2 and 3 on the otner
as
vagal fibers.
side (Fig. 2.14.2).
APPARATUS 4. Vagal escape: After recording the vagal
inhibino
continue the stimulation of vagus till the heart recovet
1. Same as in properties ol cardiac
2. Stimulating electrodes
muscle(Refer (Chapter 2.13) automatically even on continuation of vagal stimulauo
This is called
3. Signal marker.
as
vagal escape (Fig. 2.14.1B).
PROCEDURE
PRECAUTIONS
. The student should record the normal cardiogrami beuore
1. Exposure of vagosympathetic trunk:
recording the elfect of each variable.
a. Expose the heart as betore. Idenury the narrow strip 2. The
of petrohyoid muscle which runs from the base of vagosymapthetic trunk should be identified oy
anatomical landmark.
 napter 2.14 EMect of
Stimulation of Vagosympathetic Trunk and White Crescentic line on
Frogstargiogram 155

Normal Vagal Staucase

candiogram ulation WCL stimulalion
efect Normal
cardiogram
A Sgnal marker

Normal
Caraisgram
Vagal stimulation Vagal escape
(ldioventricular rhyihm)
Sgnai marker
B
Figs. 2.14.1A and B: (A) Effect of stimulation of
vagosympathetic trunk and white crescentic line (cL
(8) Phenomenon of
vagal escape.

Signal manKer Vagal stimulation

Normal Staircase
cardiogram

Fig.2.14.2:Efect of white crescentic line (WCL) stimulation. Flg. 2.14.3: [Link] after prolonged vagal
mulation.

[Link] to a higher strength of stimuli should 3. Vagal escape: When the vagus (it supplies the ventricle
be used to study the effect of vagal stimulation on
heart.
in amphibia) is stimulated, the heart at first stops, but
as the stimulation is continued, the heart escapes from
Label the recording properly to mark the start and this inhibitory effect and starts to beat once
again-a
termination of stimulation. phenomenon called vagal escape. The following factors
are involved:
a. Idioventricular rhythm: When the heart stops due to
PHYSIOCLINICALSIGNIFICANCE vagal stimulation, the ventricles start generating the
Unlike the amphibian heart, the vagal and sympathetic impulse whích is known as idioventricular rhythm. lt
nders are separate in human beings, therelore,
only is significantly lower than the normal heart rate (Fig.

Parasympathetic effects are seen

on stimulating the 2.14.3).
us On
results vagal
continuous stimulation of
the vagus, there b. Depletion of acetylcholine: On continuous stimula
escape (Figs. 2.14.2 and 2.14.3). tion of vagus, the acetylcholine released at the nerve
4Vagal tone is the tonic inhibitory influence
of the vagus
on the human heart. When the heart is denervated
endings is depleted after sometime. Therefore the
eltect of vagal stimulation on heart is temporary.
sympathetic and parasympathetic supply are cut), the
untrinsic heart rate is 100-120/min, whereas the normal
C. When the vagosympthetic trunk is
continuously
stimulated, the sympathetic fibers are also activated.
neart rate in humans is 60-100/min. This decrease in the This sympathetic effect may overpower the vagal
heart rate is due to vagal tone. effect, thus releasing the ventricle from inhibition.
156 Section 2: Amphibian (Frog) Experiments

[Link] syncope: 1f there is

sudden stimulation oBSEHVATItRESULT
a

ot vagus nerve, there occurs a transient and sudden

loss of consciousness. Strong emotions in hùmans
may lead to
vagal syncope, but the immediate vagal Descibe the efects of the following on
frog's heart
escape restores the heart beat. Vagal stimulation
(short duration)
QUESTIONS Vagai stimulation
Q.1. What type
of nerve fibers are present in the vagosympa- long duration)
thetic trunk?
Q.2. What is vagal tone?
WCL stimulation
Q.3. What is vagal escape and what is its cause?
Q.4. If both sympathetic and
parasympathetic nerves to the
heart are blocked, what is the effect on heart
rate?
Q.5. Which nervèfibers constitute the white crescentít line?
Q.6. Describe the autonomic
supply of mammalian heart. INTERPRETATION
Mammalian heart is supplied by both
sympathetic
and
parasympathetic nervous system. The vagi supply fibers to *****************"************************************
the sinoatrial (SA) node, atrioventricular
(AV) node and atrial ***** ***"**********************************************************************
muscles. No vagal fibers are distributed to ventricles. In some
individuals, right vagus is dominant, whereas in others it is ****************

the left vagus.

****************

The sympathetic fibers supply SA and AV node and

*********

********************************************
muscles of atria and ventricles.
*********************
**********************s**
 Chapter 2.14: Effect of Stimulation of Vagosympathetic Trunk and
White Crescentic Line on
Frog's Cardiogram
STUDY NOTES

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 Date. .*****************

CHAPTER2.1
Effect of Variables on
Intact Frog's Heart

AIM influx and, therefore, the torce of

contraction
To study the effect of variables intact
(negative inotropic effect).
on
frog's heart b. Nicotinic receptors are present on motor
end plates
THEORY (blocked by curare, etc.) and on
neurons in
postganglionic
autonomic ganglia.
1. Adrenaline: (at autonomic Hexamethonium
Itis
ganglion) and tubocurarine (at
a.

b.
a
sympathomimetic agent. neuromuscular junction) block these
receptors.
It actson both a and 3. Atropine:
f receptors on the
target organs.
C. In the heart it acts on
have
ß, receptors which on stimulation a. tis a
parasympatholytic agent.
b. It blocks the action of
positive chronotropic and inotropic effect.
a
In ACh on the muscarinic
the cardiac muscle cells, it increases the receptors.
mainly to Ca"" ions and, to some extent Na*permeability
ions. Influx
c. When
applied on the heart after ACh, it has no
of Na in the effect,
pacemaker cells increases the but when applied before ACh, atropine blocks
phase of the action potential (AP) thus slope
of the
4
inhibitory action of ACh.
the heart rate. The
large influx of Ca** increases the increasing 4. Nicotine:
force of contraction.
2.
Acetylcholine (ACh): Acetylcholine acts on two types of
a. It acts
through the nicotinic cholinergic receptors
which are present in the
receptors: and in the neuromuscular end plates
a. Muscarinic (M,) receptors are b. When
peripheral [Link]
muscle, smooth muscle and present
on cardiac applied to the frog's heart
it acts on the
exocrine
including sweat glands. These are blocked glands parasympathetic ganglia located on the white
by the crescentic line.
drug atropine. C. In
ACh acts
directly the
low
concentrations, nicotine stimulates tne
increases the
on
pacemaker cells and postganglionic parasympathetic fibers causin8
more ol these ions
permeability K ions which causes
to
bradycardia, while in high concentrations it bloCkS
to

hyperpolarization
move out. This results the
ganglia by causing persistent
depolarization.
in

phase which
AP thus decreases
4 of the the
slope o APHANATUS
(negative chronotropicdecreasing
effect). the heart rate
When ACh
increa the directly on atria and ventricle
acts 1. Same in
as
recording of normal
cardiogram (KeJe
shortens permeability to K* ions; the K'
it
efflux
Chapter 2.12).
phase 2 ofAP which decreases the 2.
Adrenaline: 1 in
Ca 10,000
3.
Acetylcholine: 1 in 100,000
 159
Chapter 2.15: Effect of Variables on Intact Frogs Heart
ropihe 0.5, drops of 0.5% n i c o t i n e solutinn on the heart Note that

there is no effect
S t i m u l a t e the Vagosynpathetic trunk and compare

PROCEDURE with the etlect ohtained betore adding nic otine

Stimulate the white rescentie Tine (We.)and
1Effect of adrenanne: ecord tew normal beats. Stop
drum and
pour a tew drops ot T in l0,000 solution
the

ot adrenaline on t h e heart. Record the effect.

10. Effect of atropine: Wash the heart with ng
solution. When normal beats are restored, pour
Stop the drum and wash the heart with Ringer's is n o effect o n
solution till the normal cardiogram reappears. atropine o n the heart. Note that there
the heart.
Effect of
then
acetyichoine: Kecord
pour a tew drops o t in
a lew normal beats
100,000 solution of
11. Stimulate the vagosympathetic
trunk and
compare
with the effect obtained before adding nicotine.
acetylcholine. Record the effect. 12. Stimulate the WCL and similarly compare the effect
A s the heart is beatnng. apply 0.5% atropine solution.
(igs. 2.15.1A to C).
There will be no ettect.

5. Stop the drum. wash with Ringer's solution.

PRECAUTIONS
6. Now study the ettect ot acetylcholine after applying
atropine soltution on the heart-the heart will not be 1. The student should record the normal cardiogram before
inhibited this time. recording the effect of each variable.
Effect of nicotine: After washing with Ringer's 2. The frog's heart should be rinsed with amphibian
solution when normal beats are restored, pour a few Ringer's solution between applications of various drugs.

1 Normal Normal
Normal ofmal Normal
al sti AGN Vagal with ACh WcL with ACh
vagal sumiulauon WCL stimuiation

Signalmarker
Aime marker

Normal Norma WCL

1 wtn
adrenalinee
0.5% atropine
vagus with
WChwith
Adrenallne
Vagus with
adrenaline
atropine atropine

Signal marker

B ime marker

MAAMDMOWIW Normal

Nicotine
WCL
Signal marker Vagus

Figs. 2.15.1A to C: Effect of adrenaline, acetylcholine, nicotine and atropine on Frog's heart.
(WCL: white crescentic line; ACh: acetylcholine)
160 Section 2: Amphibian (Frog) Experiments

a. Stimulation of vagosympathetic trunk will not

QUESTIONS inhibition of the heart.
ot
ause

Q.1. What the locations in the b. WCL stimulation will cause inhibition of the heart.
are
body where acetylcholine
is released? Q6. Explain how transient depolarization of neuron excites i
Acetylcholine is released at the following sites: wheres s rsistent depolarization inactivatesit?
.Preganglionic sympathetic nerve endings (in the ganglia). Transient
depolarization drings the resting membran
potential toward the firing levels, whereas the persistent
Preganglionic
endings.
and postganglionic parasympathetic nerve
depolarization inactivates the Na channeis thus preventing

Postganglionic sympathetic fibers supplying the sweat the generation of action potentials.
glands, pilomotor muscles and those supplying the blood
vessels of skeletal
muscles are cholinergic.
0BSERVATI9H AND RESULT
Neuromuscular junctions of all skeletal muscle fibers.
Many synapses in the CNS.
Q.2. How is acetylcholine inactivated in the body? What are
Variables Heart rate Force
of contraction
anticholinesterases?
The enzyme acetylcholinesterase (AChE) catalyzes the Adrenaline
hydrolysis of ACh to holiFe and acetate, which are reused.
Anticholinesterases are drugs which inhibit the action or Acetylcholine
AChE, so that ACh is preserved at the site for a longer time.
These drugs include physostigmine (eserine), neostigmine Atropine
and
diisopropylfluorophosphate (DFP), an ingredient of
some pesticides and nerve gases. Neostigmine is used in the Nicotine
treatment of
myasthenia gravis.
Q.3. What is the effect of local application
of adrenaline,
nicotine and atropine on the frog's heart?
Q4. Name the agents that block the
cholinergic receptors?
Q.5. What is the effect of stimulation of
INTERPRETATION
and WCL following the
vagosympathetic
trunk
application of atropine and 0.5%
nicotine? **********"************"************ *********************

After application of 0.5%

atropine there is blockade of the
muscarinic receptors located on the cardiac muscle. Thus *******************************"*************************************************
stimulation on the vagosympathetic trunk and WCL will be
ineffective. ***********"******************************************************************** *********

About 0.5% nicotine when

applied to frog's heart acts on
the parasympathetic *** ******"****************************************************
ganglia located on the WCL. Thus after
application of nicotine:
****************************
*********************************** *
 Chdpter L.1D: t e c t o van

STUDY NOTES
*******************************************

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 Dat *.**********

CHAPTER 216
Perfusion of Isolated Frog's Heart

AIM
able osition of the amphibian Ringer-Locke's
To study the effect of variables on isolated frog's heart. 30lutio edfor perfusing the isolated heart of frog.
NaCI 0.60 g
THEORY CaCl, 0.012 g
The rate and force of KCI 0.014g
contraction of heart is affected
by different drugs and ions. These NaHCO, 0.01 g NaHCco
change the ionic
composition of the myocardial cells and Na HPO, 0.001 g dissolved before CaCI, is added
either acting directly on ion channels
the
nodal tissue Glucose 0.1 g (to be added
or on different just before use)
receptors. ln this practical, the effect of various drugs and E d water o make it 100 mL
chemicals are being observed in isolated
frog's heart. (NaC: sodium chloride;
CaCl; calcium chloride; KCl: potassium
NaHCO,: sodium bicarbonate; Na HPO; disodium chloride;
APPARATUS phosphate)
1.
Amphibian Ringer-Locke's solution on a stand and fix the cannula in
the heart lever. Connect
a clamp directly above
2. Mariotte (perfusion) bottle the side arm of the cannula to
reservoir
3. Symes cannula a

the
containing Ringer-Locke solution and raise
4. 1%
CaCI, 1% NaCl and 1% KCI reservoir about 30 cm above the heart
5. Adrenaline: 1:100,000 pressure head (Fig. 2.16.1).
to provide a
6. Acetylcholine: 1:1000,000. Push the bent pin of the lever
ventricle, make necessary
through the apex of the
Composition of Amphibian Ringer-Locke Solution beats. adjustments and record lew a

The fluid 4. After the

required for
perfusing the isolated tracing has stabilized, raise the reservoir to
has the
composition given in Table 2.16.1. heart of frog increase the perfusion
5. Add 1% NaCl solution pressure.
via the side tube of the
PROCEDURE record its efects. Wash the cannulaain
heart with the perfusion llud
and study the effects of 1%
KCl and 1% CaCl, solutions.
1.
Expose the frog's heart and remove the 6. Wash the heart
well with
athread around the sinus.
Make a smallpericardium.
Pass and study the effects of Ringer-Locke solution
introduce a Syme's cannula into it slit in the sinus, adrenalin and acetylcholine
2. and tie a knot (Fig. 2.16.2 and Table 2.16.2).
Cutthe aorta and other around it.
with the heart.
Fit
vessels and lift the cannula
along NOTE
a
Starling's heart lever
upside down
In this
case, the
upstroke is systole and the downstroke is diastole
 Heart 163
Chapter 2.16: Perfusion of Isolated Frog's

Perfusion
(Mariotte)
Syme's cannula bottle

Recording drum Ventricle

Lever

Beaker

Fig.2.16.1: Perfusion of frogs heart.

Normal
Normal 1% CaCl, (1 mL) 1% KCI (1 mL) NOrma 1% NaCi (1 m

ime marker

Normal Normal Normal

Adrenaline ACh
(0.5 mL) (O. mL

1imemarker
Fig.2.16.2: Effect of variables on isolated frogs heart preparation.

Table 2.16.2: Effect andnanism of variables on isolatedfrog's heart

Agent Heart rate Force of contraction

[Link] (196) No effect There may bea decrease ih the contractility as Na" competes with
Ca
2KC196) because of decrease in RMP of nodal Finally the heart stops in diastole. This is because of decrease in RMP of

OtehkOissue atrial and ventricular muscles

[Link], (196) No effect Increase in ECF Ca" leading to increase in intracellular Ca". Marked3
increase in Ca" in EE may cause calcium rigor
*. Adrenaline Increases, due to increased Na* entry Increases due to Cas" ijflux in myocardial fibers

in nodal tissues
.Acetylcholine Decreases, HR as K' efflux Decreases because of decrease in ca" intiUx in myocardial fibers
nyperpolarizes the nodal tissues

(ECF: extracellular fluid; RMP: resting membrane potential)

QUESTIONS
[Link] the effets of various drugs and chemicals on frogs heart.
What do you mea uy calcium igor