Effects of Different Ion Concentrations on the Contraction of the Striated

Muscles of the Frog

Estareja, Engemar C.
Biology Student, Department of Biology, College of Science Polytechnic University of the Philippines
Abstract
Muscle is one of the most important organ in the body that accounts for the most essential
function in the maintenance of movement in a biological organism. These muscles have different
properties, suitable to produce movement and enables tension made by contraction and relaxation by
which specific ion interactions play a crucial role in making such movements possible. This study aims to
determine the effect of different ions on muscle contraction and observe the structural changes on the
striated muscles upon subjection to the different ion solutions.
Muscle strips was obtained from the striated muscle of the frog and each slide containing the
strips were bathed in the different solutions, slide A was NaCl, slide B was KCl, in slide C was CaCl2 and
slide D (control) was Ringers solution. The length of the muscle strips were measured before and after
adding the follow-up solutions for the computation of percent of contraction. In muscle response to salt
solutions, one hind leg and heart of the frog were removed and placed on a separate petri dish containing
saturated KCl solution. Heart soaked in saturated solution of Calcium chloride has the longest contraction
time same applies to hind legs that were soaked in Calcium chloride solution. After conducting the activity
it can be concluded that different ions or particular ions such as potassium, sodium and calcium has a
specific interaction within the skeletal and cardiac muscle contraction and relaxation phase.
Keywords: muscle contraction, ions, striated muscle

Introduction
Muscular movement of contraction and
relaxation is one of the most complex and crucial
physiological system in an organism. Muscles
possesses different properties suitable to
produce movement and enables tension made
by contraction (Hopkins, 2006). These properties
however must be integrated with accompanying
several processes for one contraction event to
occur.
Physiologically,
cardiac
muscle
contraction rhythm is different from the
contraction of the skeletal muscles, as these two

varies being involuntary and voluntary

respectively (Berne and Levy, 1993). As a
process, calcium ions that are stored in the
sarcoplasmic reticulum are released following
opening of ion channels upon receiving stimulus,
and binds to tropomyosin-troponin complex,
exposing the actin filaments. An ATP molecule
will bind to the myosin head, and will be
converted to ADP and Pi. The myosin then
connects to the actin forming cross bridges and
the conversion of ATP to ADP and Pi will

generate a sliding movement towards the center

of the sarcomere. When a new ATP binds to the
myosin head, the cross bridges break and the
myosin head is unattached to actin and return to
its relaxed position (Clark et. al., 2002). The
myocardium, like skeletal muscle, responds to
stimulation by depolarization of the membrane,
which is followed by shortening of the
contractile proteins and ends with relaxation
and return to the resting state. However, unlike
skeletal muscle, which shows graded
contractions depending on the number of
muscle cells that are stimulated, the cardiac
muscle cells are interconnected in groups that
respond to stimuli as a unit, contracting together

whenever a
(Harvey,2008).

single

cell

is

stimulated

These muscle contraction is greatly

affected by different ions which play an
important role in muscular physiology, from the
start of transmission of the impulse, up to the
attachment of actin molecules to myosin as it is
exposed through binding of calcium ions to
troponin (Hopkins, 2005: Brien et. al., 1993). This
study aims to determine the effect of different
ions on muscle contraction and observe the
structural changes on the striated muscles upon
subjection to the different ion solutions.

METHODOLOGY
A. Ions and Muscle Contraction
Very thin strips of ventricular muscle
were cut parallel to the direction of the muscle
fibers using scalpel and forceps. The strips were
less than 1 mm in width and anywhere in 2030mm in length. Thin muscle strips were
transferred and oriented onto each slide with
the use of forceps and 5 drops of Ringers
solution was added to cover the strips. Excess
solution was drained by tilting the slide and
wiping it with a tissue paper. The muscle length
was measured before and after subjection to the
solution and the percent contraction was
calculated using the formula given below. The
solutions are: NaCl solution for slide A, KCl for
slide B, 1:1 solution of 1 mM Cacl2 to slide C and
Ringers solution for slide D as for the control.
The Following equation was used to
determine the percent of contraction:

B. Muscle Response to Salt solutions

Ringers solution was used to wash the blood
from the removed hind legs and heart of the
frog. The hind leg and heart was placed
separately on a petri dish containing saturated
CaCl2 and KCL solution. The response of the
heart and hind leg was observed and noted on
how long the muscle responded.
RESULTS AND DISCUSSION
A. Ions and Muscle Contraction
Ringers solution was used as the constant

solution before the application of the following;

NaCl in slide A, KCl in slide B, CaCl2 in slide C, and
water in slide D. According to the study of Curtis
in 1962, that both fall in membrane potential
and resistance can be explained by assuming
that Ringers solutions increased the
permeability of the membrane to all ions and
normal range of resting membrane potential. A
thin muscle strip observed under the microscope
showed muscle fibers at its resting potential.

Length
Length after
before
exposure (in
exposure (in mm)
mm )
A (NaCl)
B (KCl)
C (CaCl2)
D (control)
E(saturated
KCl)

20.00 0.70
22.33 1.08
23.001.41
24.671.08
21.000.71

14.331.02
18.670.88
19.001.15
23.331.20
18.330.33

Table 1. Percentage Contraction of Muscle in the Varying

Solutions

The muscle strip under NaCl solution was curled

and the length was decreased significantly due
to the sudden sodium influx. The difference in
electric membrane potential inside and outside
the membrane will cause the ions to move.
Followed by calcium, calcium generally aids in
contraction by reacting with regulatory proteins
that in the absence of calcium prevent
interaction of actin and myosin (Szent - Gyorgi,
1975). KCl also induced contraction but
theoretically. KCl should help in the relaxation
state of the muscles since there would be an
efflux of potassium. As prior to the control, it
should have no muscle contraction but in the

data presented, it exhibited that the action

might due to personal error.

45
40
35
30
25
20
15
10
5
0

Figure 1. Shows the mean percentage contraction of each

muscle fiber when subjected to the different solutions.

B. Muscle response to salt solution

Muscle
Type
Heart
Hind Leg

Time
CaCl2
20 mins and
13 secs
7 mins and
4 secs

KCl
1 min and 40
secs
5 mins and 25
secs

Table 2. Response of Heart and Hind Legs in KCl and CaCl2

The heart representing a cardiac muscle and the

hind leg for a skeletal muscle were subjected in
two (2) different kinds of salt solution, but before
subjecting the heart of the frog in kind of salt
solution. Ringers solution which contains equal
amounts of salt and other substance to make the
solution neutral, was used to neutralize the

charge or the resting membrane potential of

cells in the muscles used in this activity (De mello,
1973).
Heart subjected to saturated Calcium Chloride
has the slowest contraction time (20 mins and
13 seconds) while heart subjected to saturated
Potassium Chloride solution lasted for 1 minute
and 40 seconds. Potassium is specifically needed
for voltage-gated potassium channels to work in
the outer membranes of cardiac muscle cells
(Parikh and Webb, 2012). These channels open
in response to a change in voltage and are
responsible for terminating action potentials and
contractions while initiating repolarization.
Likewise potassium ions are an important
element in all phases of heart generating action
potential. In heart contraction, during phase 0,
heart cells become less permeable to potassium
and voltage-gated sodium channels open,
producing rapid depolarization and contraction.
During phase 2, there is an increased membrane
permeability to calcium, which eventually allows
more sodium to flow into the heart cells by
which during phase 3, sodium and calcium
channels close, which leads to heart muscle
relaxation. (Pinell et al., 2007). In hind legs same
principle can be applicable, potassium is
important ion in generating action potential thus
saturated potassium chloride enable the hind
legs to contract and last its contraction time (5
minutes and 25 seconds)
Moreover during skeletal muscle contraction,
there is a phase called plateau phase which is
achieved by a balance between the influx of
Ca++ through Ca++ channels and the efflux of
several types K+ channels.
CONCLUSION
Ions greatly affect the mechanism of the

contraction of the striated muscles. Calcium

allows excitation-contraction coupling system.
Chlorine is responsible for repolarization of the
membrane. KCl is responsible for potentiation.
Water containing hydrogen and oxygen allows
the proteins that were present in the contractile
apparatus to move easily allowing contractions
to generate. The contractions of muscle are
higher on the ions that trigger action potential to
create contractions and lower on the ions that
triggers resting membrane potential to create
relaxation. Differences in the ion concentrations
inside and outside the membrane create a
significant effect on the reaction of the striated
muscles.
The observation of the hind legs must be done
immediately to see the contractions in the salt
solutions.
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