MUSCLES: STRUCTURE AND TYPES

The word muscle is derived from the Latin word musculus. Muscles are the structures formed by the bundles
of the muscle cells in the form of the fibers and possess the property of contraction on stimulation. Muscle is
unique as it can contract leading to its shortening and generating force after consumption of adenosine
triphosphate (ATP). This machine, therefore can convert the chemical energy into mechanical one (e.g.,
movement) like an engine. So, muscle is an organ specializing in the transformation of chemical energy into
movement which is essential for life. Although only a few families of proteins are responsible for movement in
the biological world, muscle has developed to optimize this function and is packed with movement-related
proteins.

Types of muscles
On the basis of the types of muscles available in the body: there are three types of muscles in the body namely
skeletal muscle, smooth muscle and cardiac muscle. Skeletal muscle makes up about 40% of the body and
smooth muscle and cardiac muscle make up nearly 10% more. All muscles have one major common
characteristic feature i.e. they can contract. The contraction of the skeletal muscles causes the external
locomotion, the contraction heart muscle causes the circulation of blood while with the smooth muscle this
causes narrowing –squeezing/ relaxation of the various viscera and movement of their content. Arteries and
veins also have smooth muscles in their walls.

1. Skeletal muscles:
The muscles attached to the skeleton (bones) are called skeletal muscles. The skeletal muscles are involved in
locomotion and day to day activities of the individual. All body movement is the result of skeletal muscle. They
are under voluntary control hence they are called voluntary muscles as well. The skeletal muscles are striated.
The fibers of the skeletal muscles are grouped into the bundles called as fasciculi. The fiber is covered by the
outer delicate areolar tissue called endomysium. The bundle or the fasciculus is covered by the layer of the
connective tissue known as perimysium. The muscle consisting of the group of the bundles is covered by the
epimysium. Most veterinary patients with the disease of the neuromuscular system exhibit abnormalities of
movement so it is important to understand how skeletal muscle functions and how it is controlled by the
nervous system.

47
 Figure: Skeletal muscle

2. Cardiac muscles:
The muscles that are present in the heart are called cardiac muscles. Among the three layers (epicardium,
myocardium and endocardium) of the heart, cardiac muscle is present in the myocardium of the heart. Cardiac
muscles alone form a special group and these muscles make the heart the wonderful automatic pump which is
used to pump blood through circulatory system tirelessly from the intrauterine (before birth) life till death of an
individual. The cardiac muscle though striated is involuntary and is not controlled by the will. Abnormalities of
cardiac and smooth muscle of the arteries and vein feature predominantly in many clinical disorders and
pharmacological mechanisms also. Therefore, we need to understand the mechanism of their contractions and
abnormalities resulted when these muscles fail to function properly.

3. Smooth muscle:
Smooth muscle forms the bulk of the wall of the internal organs. These muscles are called smooth muscles as
they look smooth under light microscope. The contractile filaments in them are not arranged in regular pattern
as seen in skeletal and cardiac muscles. Smooth muscles are present in different organs like stomach, intestines,
passages like bronchi, arteries, ureters, urethra, arteries, veins etc. These muscles are not striated and are not
under the voluntary control. Individual cells of the smooth muscles are spindle shaped. However, the structure
of the smooth muscles varies widely. In general they are formed of fusiform cells with a single nucleus but in
some places (e.g., uterus) the cells may be very long. The contractile proteins are as usual but the thick and
thin filaments are haphazardly arranged, hence the striped appearance is not seen.
The smooth muscles in different parts of the body vary widely in their physiological properties, their general
description is very difficult. Smooth muscle which lines the walls of the arteries control blood pressure, and in
the gastrointestinal tract controls the digestion of food by causing movement of the intestine. In general the
smooth muscles of the body are divided into two groups: multi-unit and single unit smooth muscle. In multi-
unit smooth muscle the individual fibers are separated electrically and contract individually; the single unit
smooth muscles have all the muscle fibers interconnected electrically by gap junctions. So, in them when a part
is stimulated the whole muscle is ultimately stimulated.

Functionally all muscles are divided into two groups:

 1. Voluntary muscles
2. Involuntary muscles

Voluntary muscles: These consist of striated muscle which generally contract on the will of own, e.g., muscles
of the extremities, head, neck, etc. Exception is the cardiac muscle, which is also striated, but is not controlled
by the will.

Involuntary muscles: These consist of non-striated muscles or smooth muscle tissues and are found in the wall
of the blood vessels, gastrointestinal tract, respiratory tract, skin reproductive tract and so on. These
involuntary muscles are not under the control of will and they are governed by the nervous input to them.

STRUCTURE OF DIFFERENT TYPES OF MUSCLES

1. Skeletal muscle
Skeletal muscle consists of a central fleshly contractile muscle belly and two tendons- one on each end of the
muscle. The muscle and its tendons are arranged in the body so that they originate on one bone and insert on a
different bone while spanning a joint. As the muscle contracts, shortening the distance between the origin and
insertion tendons, the bones move with respect to each other, bending at the joints. When activated by the
motor nerve, a skeletal muscle can only shorten. Most joints have one or more muscles on both of sides, either
to decrease its angle (flexion) or to increase its angle (extension). All movements performed by an animal are
the results of contraction of skeletal muscles across a moveable joint. It is therefore very important to
understand the anatomy and physiology of skeletal muscle and resultant moveme3nt of the body parts due to
such contractions..

Each muscle belly as seen during dissection consists of large number of muscle fasciculi, and each fasciculus is
composed of large number of muscle cells (usually called muscle fibers) that span the several inches between
the origin and inserting tendons. Each muscle fiber is multinucleated and behaves as a single unit. The skeletal
muscle fibers range between 10-80 micrometers in diameter and contain multiple mitochondria and other
intracellular organelles. The outer limiting membrane is called sarcolemma. The sarcolemma consists of a true
cell membrane, called as the plasma membrane and an outer coat made up of a thin layer of polysaccharide
material consisting of numerous thin collagen fibrils which attaches to the tendons at the cell extremities.

Each muscle fiber consists of several hundred to several thousand myofibrils arranged in parallel along its
length. Each myofibril is made up of a series of repeating sarcomeres, the basic contractile unit of the muscle
fiber. Each myofibril has, lying side by side, about 1500 myosin filaments and 3000 actin filaments, which are
largely polymerized protein molecules that are responsible for muscle contraction. The thick filaments in the
myofibrils are myosin and the thin filaments in the myofibrils are the actin. The myosin and actin filaments
partially interdigitate and thus cause the myofibrils to have alternate light and dark bands. The light band
contains only actin filaments and is called as I bands. The dark bands contain the myosin filaments as well as
the ends of the actin filaments where they overlap the myosin. The dark bands are called A bands because they
are anisotropic to polarized light. The small projections from the sides of the myosin filaments are called
cross-bridges which protrude from the surfaces of the myosin filaments along the entire extent of the filaments,
except in the very center. It is the interaction between these crossbridges and the actin filaments that causes
contraction.

The end of the actin filaments are attached to the Z discs. The Z discs are composed of filamentous proteins
different from the actin and myosin filaments. The Z disc passes from the myofibril to myofibril, attaching the
myofibrils to each other all the way across the muscle fiber. Thus, the entire muscle fiber has light and dark
bands, as do the individual myofibrils. These bands give the skeletal and cardiac muscle their striated
appearance. The portion of the myofibril that lies between two successive Z discs is called a sarcomere. In
normal condition, the length of the sarcomere is about two micrometers. The sarcomere is the basic unit of
contraction; with electrodes it is possible to stimulate a single sarcomere and make it contract. When a muscle
fiber is stretched beyond its resting length, the ends of the actin filaments pull apart leaving a light area called
the H zone in the center of the A band.

The myofibrils are suspended inside the muscle fiber in a matrix called sarcoplasm. The fluid of the
sarcoplasm contains large quantities of potassium, magnesium, phosphate, and protein enzymes. Tremendous
numbers of mitochondria lie between and parallel to the myofibrils and this large number of mitochondria are
essential to produce large amounts of ATP that is required for the contracting myofibrils. From the sarcolemma,
tubules called T- tubules arise and enter the muscle cell. These T-tubules proceed transversely forming a sort of
grid round the myofibrils. Another set of tubules, called as sarcoplasmic reticulum (SR), run parallel to the
longitudinal axis of the myofibrils. The sarcoplasmic reticulum is the extensive portion of the endoplasmic
reticulum. The T-tubules are those channels which communicate between the extracellular fluid (ECF) and the
interior of the muscle cell, and the sarcoplasmic reticulum is confined within the cell, i.e. it does not
communicate with the ECF. The T-tubules and the sarcoplasmic reticulum are responsible for muscle
contraction.
 Figure: Transverse tubule and sarcoplasmic reticulum system

Molecular characteristics of the contractile filaments

Myosin filament

The myosin forms the thick filaments in the myofibril. The myosin filament is made up of about 200 individual
myosin molecules, each having a molecular weight of about 480,000. The myosin molecule is composed of six
polypeptide chains; two heavy chains each with a molecular weight of about 200,000 and four light chains
with molecular weight of about 20,000 each. The two heavy chains wrap spirally around each other to form a
double helix. One end of these chains is folded into a globular protein mass called the myosin head. There are
two free heads lying side by side at one end of the double helix myosin molecule and the other end of the coiled
helix is called the tail. The four light chains are also the parts of the myosin heads, two to each head. These
light chains help to control the function the head during the process of muscle contraction. The tails of the
myosin molecules are bundled together to form the body of the filament and many heads of the molecules hang
outward to the sides of the body. A part of the helix portion of each myosin molecule extends to the side along
with the head thus providing an arm that extends the head outward from the body. The protruding arms and
heads together are called cross bridges and each of these is believed to be flexible at two points called hinges
one where the arm leaves the body of the myosin filament and the other where the two heads attach to the arm.
The hinged arms allow the heads to be extended either far outwards from the body of the myosin filament or to
be brought close to the body. The hinged heads are believed to participate in the actual contraction process.
There are no cross bridge- heads in the very center of the myosin filament for a distance of about 0.2 micron
because the hinged arms extend toward both ends of the myosin filament away from the center and therefore
there are only tails of the myosin molecules and no heads. The myosin filament itself is twisted so that each
successive set of cross-bridges is axially displaced from the precious set by 120 degrees. This ensures that
the cross-bridges extend in all directions around the filament.
The head of the myosin functions as an ATPase enzyme, which is essential for the muscle contraction. This
property allows the head to cleave and energize the contraction process.

Actin filament
The actin filament is composed of three different protein components; actin, tropomyosin, and troponin. The
actin filament forms the thin filament in the myofibrils. The backbone of the actin molecule is a double
stranded F actin protein molecule, which wound in a helix in the same manner as the myosin molecule. Each
strand of the double F-actin helix is composed of polymerized G-actin molecules, each having a molecular
weight of 42,000. There are approximately 13 of these molecules in each revolution of each strand of helix.
Attached to each one of the G-actin molecules is one molecule of ADP. It is believed that these ADP
molecules are the active sites on the actin filaments with which the cross bridges of the myosin filaments
interact to cause muscle contraction. The active sites on the two F-actin strands of the double helix are
staggered giving one active site on the overall actin filament approximately every 2.7 nanometers. The actin
filament is approximately 1 micrometer long and the bases of the actin filaments are inserted strongly into the Z
discs while their other ends protrude in both directions into the adjacent sarcomeres to lie in the spaces between
the myosin molecules. The actin filament also contains two additional protein strands that are polymers of
tropomyosin molecules, each of which has a molecular weight of 70,000 and each tropomyosin strand is
loosely attached to an F-actin strand and that in the resting state it physically covers the active sites of the
actin strands so that interaction cannot occur between the actin and myosin to cause contraction. Attached
approximately two thirds the distance along each tropomyosin molecule is a complex of three globular protein
molecules called troponin. The troponin protein has three globular protein subunits called troponin I,
troponin T and troponin C. The troponin I has a strong affinity for actin, the troponin T for tropomyosin
and the troponin C for calcium ions. This complex attaches the tropomyosin to the actin. The strong affinity
of the troponin for calcium ions is believed to initiate the contraction process.

2. Structure of smooth muscle

The smooth muscle has thick and thin filaments that are not arranged in sarcomere; therefore they appear a
homogenous rather than striated. The principle of contraction applies to the smooth muscle as that of skeletal
muscle. Smooth muscle can be divided into two major types: multiunit smooth muscle and single unit smooth
muscle.
a. Multi-unit smooth muscle: The multiunit smooth muscle fibers can contract independently of the others,
and their control is exerted by nerve signals. Some examples of multiunit smooth muscle found in the
body are the ciliary muscles of the eye, the iris, the nictitating membrane and the pilo-erector muscles of
the skin.

b. Single-unit smooth muscle: The whole mass of the hundreds to millions of muscle fibers that contract
together as a single unit is called as single-unit smooth muscle. The fibers are usually aggregated into
sheets or bundles, and their cell membranes are adherent to each other at multiple points so that force
generated in one muscle fiber can be transmitted to the next. These types of muscles are present in the
walls of the most of the viscera including the gut, bile ducts, the ureters, the uterus, and many blood
vessels- also called visceral smooth muscle.
3. Structure of Cardiac muscle
The cardiac muscle fibers are arranged in a latticework; the fibers divide, recombine, and then spread again. The
cardiac muscle fibers are made up of many individual cardiac muscle cells that are connected with each other in
series. These cells are separated by the intercalated discs (angulated dark area crossing the cardiac muscle
fiber), which are actually the cell membrane separating the cells. Gap junctions are present at the intercalated
disc, which allows the relatively free diffusion of ions. Cardiac muscle, like skeletal muscle has a striped
appearance. These cross- striations have the same structural basis in cardiac and skeletal muscle. Each
striated cardiac muscle fiber is made up of a few hundred myofibrils. Each myofibril has a repetitive pattern of
light and dark bands. The alignment of these bands in adjacent myofibrils accounts for the striated appearance
of the whole muscle fiber. Each repeating unit of the myofibrilar bands is called a sarcomere. Like in skeletal
muscle, each cardiac muscle sarcomere is composed of an array of thick and thin filaments. The thin filaments
are attached to the Z discs, and they interdigitate with the thick filaments. The thin filaments are composed of
actin molecules. The thick filaments are composed of myosin molecules. Each myosin molecule has a flexible
end (myosin head) that protrudes from the thick filament. The myosin head can bind reversibly to one of the
actin molecules in the nearby thin filament.
 Table: Comparison of skeletal, cardiac and smooth muscle

Skeletal Cardiac Smooth

Property
Muscle Muscle Muscle
Striations Yes Yes No
Relative speed
Fast Intermediate Slow
of contraction

Voluntary control Yes No No

Membrane
Short Long
refractory period

Nuclei per cell Many Single Single

Nerves,
Control of Beats spontaneously
Nerves Hormones,
contraction but modulated by nerves
Stretch

Cells connected by
intercalated Discs or No Yes Yes
gap junctions
 MEMBRANE PHYSIOLOGY

Body fluid
The fluid present in body regulates different physiological functions. There is considerable variation in body
water content between different breeds, ages, sexes, nutritional status and various other conditions of an animal.
The body water in a lean non herbivorous animal of 70 kg body weight is approximately 50 liters. Fluid
contained within cells is called intracellular fluid. All fluid lying outside of cells is called extracellular fluid.
Water molecules can rapidly penetrate most cell membranes.
Intracellular fluid compartment
Fluid present inside the cell are collectively called as intracellular fluid (ICF). The ICF compartment forms
about 50% of the body mass. In humans 25-40 liter fluid is present in approximately 75 trillions of cells of the
body.
Extracellular fluid compartment
All fluid lying outside the cells is called the extracellular fluid (ECF). The ECF forms about 20% of the body
mass. The total amount of fluid in extracellular compartment averages about 15 liter in 70 kg man. The
extracellular fluid is divided by the walls of the vascular system into the interstitial fluid and the plasma.
Roughly an amount of water equal to 50% of the body weight lies within the cells, 15% in the interstitial spaces,
and 5% in blood plasma. Water in the alimentary canal is also included in extracellular fluid. Similarly,
cerebrospinal fluid, aqueous humor of eye, synovial fluids, peritoneal fluid, pericardial fluid, pleural fluid,
urine, and bile are special type of extracellular fluids, so called transcellular fluids since these fluids are
somewhat separated from the main body of the extracellular fluid.

Total body fluid (40 liter)

Extracellular fluid

15 liter

Intracellular fluid
cell

Blood volume (5 liter)

Red blood

25 liter
(3 liter)

(2 Liter)
Plasma

The body fluids diagrammed showing the extracellular, intracellular, blood volume and the total body fluids
in humans.
Constituents of Extracellular and intracellular fluids
The constituent of extracellular and intracellular fluid is shown in the table below. From this table we can learn
that these fluid in ionic form exert osmotic pressure - the extracellular fluid contains large amount of sodium
and chloride, reasonably large amount of bicarbonate, but small quantities of potassium, calcium, magnesium,
phosphate, sulphate and other organic acids. Also, plasma contains some protein while the interstitial fluid has
very little.

The intracellular fluid on the other hand, contains small amount of sodium and chloride and almost no calcium.
It, however, contains large quantities of potassium and phosphate and moderate amounts of magnesium and
sulfate. Intracellular fluid also contains relatively large amounts of proteins.

Table: Osmolar substances in extracellular and intracellular fluids

Constituents Plasma (mOsm/lit) Interstitial (mOsm/lit) Intracellular

(mOsm/lit)
+
Na 143 140 14
K+ 4.2 4.0 140
++
Ca 1.3 1.2 0
Mg++ 0.8 0.7 20
-
Cl 108 108 4
-
HCO3 24 28.3 10
--
HPO4 2 2 11
SO4 0.5 0.5 1
Glucose 5.6 5.6 4
Protein 1.2 0.2 4
Urea 4 4 4
Creatinine 0.2 0.2 9
Amino acids 2 2 8

Table: The non-electrolytes of the plasma

Constituents mg/dl
Phospholipids 280
Cholesterol 150
Neutral fat 125
Glucose 100
Urea 15
Lactic acid 10
Uric acid 3
Creatinine 1.5
Bilirubin 0.5
Bile salts traces
Structure of biological membrane
Biological membrane is composed of lipoprotein complexes with a bilayer of phospholipid molecules
oriented perpendicular to the plane of the membrane. Cell membranes are approximately 80 A0 thick.
Phospholipids are the molecules that have two long tails of hydrophobic fatty acid and a head containing a
charged, hydrophilic phosphate group. So, the outside surfaces of biological membranes are hydrophilic
and inside surfaces are hydrophobic lipid bilayers which prevent the free entry of water and many solutes.
Biological membranes, thus, behave as if they were lipoids punctured by aqueous pores. The polar groups i.e.
hydrophilic groups (glyceryl phosphate attached to ethanolamine/choline or hydroxyl group of cholesterol) of
the bilayers are oriented at both surfaces and point outward to hydrogen bond with water while the oily, fatty
acid tails point inward toward one another and away from the water. The long nonpolar hydrocarbon chains
are embedded in the matrix, with adsorbed extrinsic and intrinsic protein molecules. The proteins are able to
float freely through the membrane are extrinsic proteins and those embedded in the lipid bilayer are intrinsic
proteins. Aqueous channels appear to be present in the central axes of the globular intrinsic (integral) proteins.
These protein channels provide pathways for passage of specific substances through the membrane. The other
type of protein associated with cell membrane is carrier protein. Conformational changes occur when the
carrier proteins bind with substances that are to be transported and then the substances move through the
interstices of the molecules to the other side of the membrane. Both the channel proteins and the carrier proteins
are highly selective in the type of molecules or ions that are allowed to cross the membrane. Glycoproteins or
glycolipids are formed on the surface by attachment to polymeric sugars, amino-sugars or sialic acids.

So, all biological membranes are a mosaic of proteins embedded in a phospholipid bilayer whether the
membrane is the outer plasma membrane separating cytoplasm from extracellular fluid or the membrane
surrounding intracellular fluid or the membrane surrounding intracellular membranous organelles such as
endoplasmic reticulum, golgi apparatus, mitochondria, lysozomes, etc. The lipid of the membrane is not
miscible with either extracellular fluid or the intracellular fluid, and thus provides a barrier preventing free
movement of water and water soluble substances from one cell compartment to other. Therefore, the lipid
bilayer is a major barrier impermeable to the usual water soluble substances as ions, glucose, and urea and
permeable to the fat soluble substances as alcohol, oxygen, carbon dioxide, etc.

Transport methods

Diffusion (passive transport)

It is random molecular movement of substances either through intermolecular spaces in the membrane or in
combination with carrier proteins. In diffusion there is net movement of molecules or ions from regions of high
concentration to low concentration. The energy that causes diffusion is by the energy of the normal kinetic
motion of the matter. The process of diffusion does not saturate as the concentration or gradient changes. There
is no interference between different substances during diffusion (no competition). The net flux (amount of
movement) is proportional to the concentration difference and the permeability of any barrier like a
membrane. Diffusion can occur through tight junctions or within bulk solutions. Diffusion of water down its
concentration gradient is called osmosis. Diffusion through the cell membrane includes simple diffusion and
facilitated diffusion.

Simple diffusion
Simple diffusion means the molecular kinetic movement of molecules or ions through a membrane opening or
intermolecular spaces without the necessity of binding with carrier proteins in the membrane.

The rate of diffusion is determined by:

a. the amount of substance available
b. the velocity of the kinetic motion
c. the number of the openings in the cell membrane through which the molecules or ions can move.

Simple diffusion can occur through the cell membrane by two pathways:
 a. Through the interstices of the lipid bilayer: The highly lipid soluble substances like oxygen, nitrogen
and alcohols travel through this route. The rate of diffusion is directly proportional to their lipid
solubility. For example, the lipid solubility of oxygen, nitrogen and alcohols are very high so that all
these can dissolve directly in the lipid bilayer and diffuse through the cell membrane in exactly the same
manner that diffusion occurs in a watery solution.

b. Through watery channels in some of the transport proteins: The protein channels are watery
pathways through the interstices of the protein molecules. Most of the protein channels are highly
selective for the transport of one or more specific ions or molecules. For example sodium channels are
very specific channels and are about 0.3 by 0.5 nm in size. The inner surface of these channels are
strongly negative and these strong negative charges pull the sodium ions more than they pull the other
physiologically important ions into the channels because the ratio of pulling force to ionic diameter is far
greater for sodium than for the others. Once in the channel the sodium ion then diffuses in either
direction according to the usual law of diffusion. So, the sodium channel is specifically selective for the
passage of sodium ions. Another set of protein channels is selective for potassium transport. These
channels are slightly smaller than the sodium channels, only 0.3 by 0.3 nm but they are not negatively
charged. Therefore, no strong attractive force is pulling ions into the channels.

Gating of the protein channels

Gating of protein channels provides a means for controlling the permeability of the channels. It is believed that
the gates are actual gate - like extensions of the transport protein molecule, which can close over the
opening of the channel or can be lifted away from the opening by a conformational change in the shape of the
protein molecule itself. The opening and closing of gates are controlled in two principal ways-
1. voltage gating
2. ligand gating

1. Voltage gates
These types of channels open or close as per the change in the electrical potential of the cell membrane. For
instance, when there is strong negative charge in the inside of the cell membrane, the sodium gates remain
tightly closed. On the other hand, when the inside of the membrane loses its negative charge these gates open
suddenly and allow tremendous quantities of sodium to flow in through the sodium channels. The potassium
gates also open when the cell membrane becomes positively charged inside the cell. However, this response
is different from that of the sodium gates for they open much more slowly the resulting outflow of potassium
ions from the cell helps in the recovery of the nerve fiber membrane at the ends of the action potential.

2. Ligand gates
Some protein channel gates are opened by the binding of another molecule with the protein, thus causing a
conformational change in the protein molecule that opens or closes the gate. This is called ligand gating and
the substance that binds is called the ligand. One best example of the ligand gated channels is the effect of
acetylcholine on the acetylcholine channels. This opens the gate of this channel providing a pore of about 0.65
nm in diameter that allows all molecules and positive ions smaller than this diameter to pass through. This gate
is exceedingly important in the transmission of signals from one nerve cell to another and from nerve cells to
muscle cells.

b. Facilitated diffusion:
Facilitated diffusion requires the interaction of the molecules or ions with a carrier protein that aids its
passage through the membrane. It is also called carrier mediated diffusion because a substance transported in
this manner usually cannot pass through the membrane without a specific carrier protein helping it. So, the
carrier protein facilitates the diffusion of the substance to the other side. Facilitated diffusion differs from the
simple diffusion through an open channel in the following very important way: whereas the rate of diffusion
through an open channel increases proportionately with the concentration of the diffusing substance, in
facilitated diffusion the rate of diffusion approaches a maximum called V max as the concentration of the
substance increases. The molecule to be transported enters the channel and then binds with the receptor on the
protein carrier. Then in a fraction of second a conformational change occurs in the carrier protein so that the
channel now opens to the opposite side of the membrane. The rate at which the molecules transported by this
mechanism can never be greater than the rate at which the carrier protein molecule can undergo
conformational change back and forth between its two states. Glucose and amino acids are transported by
facilitated diffusion.

Figure: Facilitated diffusion through ligand gated channel

Ion channels
There are ion channels for K+, Na+, Ca2+, and Cl-, and each exists in multiple forms with diverse properties. The
+ +
K channel promote the efflux of K ions out of the cell; similarly the sodium channel promotes the influx of
the sodium inside the cell. The channel proteins functions as the leak channels and allow for the entry or exit of
the different ions from the cell.

II. Active transport

Active transport means movement of ions or other substances across the membrane in combination with a
carrier protein but additionally against an energy gradient. It is also called uphill transport. Such
movement of substances from a low concentration to a high concentration requires energy beside kinetic
energy that causes the movement. Among the different substances that are actively transported through cell
membranes are sodium ions, potassium ions, calcium ions, iron ions, urate ions, several different sugars and
the amino acids. Active transport depends on transport of substances by carrier proteins that penetrate through
the membrane, the same as is true for facilitated diffusion. However, in active transport, the carrier functions
differently from the carrier protein must now impart energy to the substance that is being transported to move it
against an electrochemical gradient. The energy for this purpose is derived from ATP. Furthermore, the active
transport carrier proteins have ATPase activity which can cleave ATP to ADP or AMP with the release of the
energy from the high energy phosphate bonds.

Active transport is of two types according to the source of the energy used to cause the transport as primary
active transport and secondary active transport. In primary active transport, the energy is derived directly
from the breakdown of adenosine triphosphate (ATP) or other energy rich substances. But in secondary active
transport, energy is derived secondarily from ionic concentration gradients that have been created in the first
place by primary active transport. In both of the above transport system, the carrier proteins that are situated in
the cell membrane functions for the transport of the substances.

1. Primary active transport:

In primary active transport, the energy is derived directly from the breakdown of adenosine triphosphate
(ATP) or other energy rich substances. Different ions that are transported by the primary active transport
include sodium, potassium, hydrogen, chloride and other ions.

a. Sodium-potassium pump
The cytosol of animal cells contains a concentration of potassium ions as much as 20 times higher than that in
the ECF. Conversely, the ECF contains a concentration of sodium ions as much as 10 times greater than that
within the cell. These concentration gradients are established by the active transport of both ions. And, in fact,
 + +
the same transporter, called the Na -K ATPase, does both jobs. The most ubiquitous active transport
mechanism in the body is the one that transports sodium ions out of cells to the exterior and at the same time
pumps potassium ions from the outside to the inside through the cell membrane, this is called the sodium-
potassium pump. This pump is present in all cells of the body and is responsible for maintaining the sodium
and potassium concentration differences across the cell membrane as well as for establishing a negative
+
electrical potential inside the cells. It uses the energy from the hydrolysis of ATP. The crucial roles of the Na -
+
K ATPase are reflected in the fact that almost one-third of all the energy generated by the mitochondria in
animal cells is used just to run this pump. This pump consists of a complex of two globular proteins a larger
one with a molecular weight of 100,000 and a smaller one with a molecular weight of 45,000.
Though the function of the smaller one is unknown, the larger protein has three specific functions:
a. It has three receptor sites for binding sodium ions on the protein that protrudes to the interior of the cell.
b. There are two receptor sites for potassium ions on the outside.
c. The inside portion of this protein adjacent to or near to the sodium binding sites has ATPase activity.

When three sodium ions bind on the inside of the carrier protein and two potassium ions on the outside,
the ATPase function of the protein becomes activated. This then cleaves one molecule of ATP, and release
high level of energy. This liberated energy is used to make conformational change in the carrier molecule and
move the sodium ion to the outside and potassium ion to the inside of the cell. If there is absence of this pump
then huge amount of positively charged substances are stored inside the cell causing lot of water holding inside
the cell, which may lead to the rupture of the cell.

Figure: sodium-potassium pump

b. The calcium pump

Calcium ions are normally maintained at extremely low concentration in the intracellular cytosol, and are
about 10,000 times less than that in the extracellular fluid. This level of calcium in the two fluid compartments
is maintained by the two sets of calcium pump. One set of the calcium pump is situated in the cell membrane
and it pumps calcium ions to the outside of the cell. The other set is situated in one or more of the internal
vesicular organelles of the cell as in the sarcoplasmic reticulum, mitochondria, etc. and it pumps calcium from
the cytosol to the organelles. In performing the above work it requires energy which is provided by the cleavage
of the ATP by ATPase activity in the carrier proteins.

Secondary active transport

When sodium ions are transported out of cells by primary active transport, a very large concentration gradient
of sodium usually develops outside the cell and very low concentration inside. This gradient represents a
storehouse of energy, because the excess sodium ion outside the cell always attempt to diffuse to the interior.
Under the appropriate conditions, this diffusion energy of sodium can literally pull other substances along with
the sodium through the cell membrane. This phenomenon is called as co-transport. This process uses proteins
similar to those for facilitated diffusion and it can couple the movement of several different molecules in each
cycle. Example: sodium cotransport of glucose and amino acids occurs especially in the epithelial cells of the
intestinal tract and renal tubules to aid in the absorption of these substances into the blood. The process
saturates when substance reaches to high concentrations due to lack of available protein. Co-transport moves
two or more molecules in the same direction across the membrane as sodium and glucose or amino acids.

In counter transport, molecules move in opposite direction using the channel proteins. A receptor site is
present on the outside of the carrier protein for sodium but on the inside for the other substance and movement
of the sodium down its energy gradient to the inside of the cell causes the other substance to be transported to
the exterior. High concentration of sodium outside the cell provides electro gradient for the movement of
sodium ion inside the cell and in doing so, it has to get exchanged with other substances as calcium or
hydrogen which is taken out of the cell. Normal active transport (Na-K ATPase) makes a strong Na gradient,
which in turn powers many secondary active transport mechanisms.

MEMBRANE POTENTIAL, RESTING MEMBRANE POTENTIAL AND

ACTION POTENTIAL

The fluids inside and outside the cells are electrolytic solutions containing 150-160 mEq per litre of positive
ions and the same concentration of negative ions. Generally, a very minute excess of negative ions (anions)
accumulate immediately inside the cell membrane along its inner surface and an equal number of positive ions
(cations) accumulate immediately outside the membrane. The effect of this is the establishment of a membrane
potential between the inside and outside of the cell. So, the cell membrane has many electrical properties. It
shows conductance, capacitance, resistance, etc. like ordinary conductors. But the cell membrane differs from
an ordinary nonliving conductor in many ways. The electrical changes in a cell membrane are usually due to
active participation of the membrane components and these components must be part of a living cell. If the cell
dies then all these properties are lost. And every cell in the body possesses some electrical potential and some
cells possessing the electrical potentials are excitable, e.g., nerve and muscle cell.

Membrane potential (MP) is the potential difference between inside and outside of the membrane of a living
cell. Membrane potential plays significant role in different functions of cells. In almost all cells, there is a
potential difference. The potential difference is in the range of -10 mV to -90 mV. The negative sign indicates
the polarity of the inside of the membrane (a value say -70 mV means that the interior of the cell is negative by
-70 mV with respect to outside, a value of +35 mV means the inside is positive by 35 mV. If the cell is in
resting condition, the membrane potential at that time is called resting membrane potential (RMP). The
membrane potential can be measured by putting micro electrodes, one inside and other outside the cell
membrane and then connecting them to a cathode ray oscilloscope (CRO). Cathode ray oscilloscope is a
highly sensitive instrument used to detect and measure bio-potentials (membrane potential, action potential)
from different sources.

The two basic means by which membrane potentials can develop are:
a. Diffusion of ions through the membrane as a result of ion concentration differences between the two
sides of the membrane, thus creating an imbalance of negative and positive charges on the two sides of
the membrane.
 b. Active transport of ions through the membrane, thus creating an imbalance of charges.

Membrane potential (MP) caused by diffusion

+
It is very important to remember that there is more K inside the cell than outside. The cell membrane at rest
is very permeable to K+ ions but not to any other ions. Because of the large potassium concentration gradient
from the inside to outside there is a strong tendency for potassium ions to diffuse outward. So, as K + diffuses
out along its concentration gradient they carry positive charges to the outside the membrane and therefore the
inside of the cell becomes negative, thus creating a state of electro-positivity outside the membrane and electro-
negativity on the inside because of the negative anions that remain behind which do not diffuse outward along
with the potassium. The anions inside the cells are mostly large molecules like proteins, phosphates, etc. which
cannot pass out. The chloride ion is permeable but cannot pass out against its concentration gradient, so it
+
cannot influence the negativity. Na is impermeable to cell membrane at rest, hence the inside remains
+
negative. When this negativity of inside becomes of sufficient strength, it draws K in and a situation is reached
+ +
when the number of K ions moving out down the concentration gradient becomes equal to the number of K
coming in due to electrical, i.e., an equilibrium is reached. This potential is also called potassium equilibration
potential. At this moment, the inside of the cell is always negative with respect to outside and the potential
difference is about -90 mV.

Membrane potentials are determined primarily by three factors:

a. Concentration of ions on the inside and outside of the cell
b. Permeability of the cell membrane to those ions
c. Activity of electrogenic pumps (e.g., Na+-K+ATPase and Ca++ transport pumps) that maintain the ion
concentrations across the membrane.

Relationship of the diffusion potential to the concentration difference- the Nernst equation
The potential level across the membrane that will exactly prevent the net diffusion of an ion in either direction
is called the Nernst potential for that ion. When a concentration difference of a single type of ion across a
membrane causes diffusion of ions through the membrane thus creating a membrane potential, the magnitude of
the potential inside the membrane versus the outside is determined by the ratio of the tendency for the ions to
diffuse in one direction or the other direction, which is determined for positive ions by the following formula at
normal body temperature of 370C:
Conc. inside EMF (Millivolts)
= ± 61 log -----------------------
Conc. outside

So, when the concentration of positive ions on the inside of a membrane is ten times that on the outside, the log
of 10 is 1 and the potential difference calculates to be -61 mV. This equation is called the Nernst equation and
the developed potential is the Nernst potential. For the Nernst [potential to develop as a result of diffusion the
membrane must be selectively permeable allowing a single type of ion to diffuse through the channels while all
other ions do not diffuse and there must be concentration gradient for the diffusible ions between the two sides
of the membrane.

The normal concentration of sodium ions inside the nerve membrane is approx 14 mEq and outside approx. 142
mEq. Thus, the ratio of these two is 0.10 and the logarithm of 0.310 is - 1.00. Multiplying this by -61 millivolts
gives a Nernst potential for sodium of +61 millivolts inside the nerve fiber membrane.

The normal concentration of potassium ions inside the nerve fiber is approx 140 mEq per litre and 4mEq per
liter on the outside. The ratio of these two is 35. So, using the Nernst equation the Nernst potential of the
potassium ions will be -94 millivolts inside the membrane. When the membrane is permeable to several
different ions, the diffusion potential that develops depends on three factors:
a. The polarity of the electrical charge of the each ion
b. The permeability of the membrane (P) to each ion
 c. The concentrations (C) of the respective ions on the inside (i) and outside (o) of th4e membrane.

Thus, the following formula so called Goldman Equation is used to calculate the membrane potential on the
+ +
inside of the membrane when two univalent positive ions sodium (Na ) and potassium (K ) and one univalent
-
negative ion, chloride (Cl ) are involved.
+ + + + - -
CNa iPNa + CK iP K + CCl OPCl
EMF (mV)= ± 61 log -----------------------------------------------------------

CNa+o P Na+ + CK+i P K+ +CCl-i P Cl-

Maintenance of the membrane potential

+ + - +
Na , K , and Cl all can move across the membrane at rest but at different rates. K can pass easily through its
leaky channels which are always open. The strong and persistent tendency of K efflux makes the inside of the
membrane negative. There is no matched efflux of the intracellular anions (they are mostly very large particles
and Cl- has minimum effect). The tendency of Na+ is to enter the cell. This is firstly because the inside is
+ + +
negative and Na is a positive ion and secondly, the Na in ECF is many times higher than the Na in ICF.
Both the concentration and electrical gradients therefore are directed inwards. But Na+ cannot pass though the
leaky channels because of its higher effective diameter along with the water molecules attached to it. The
voltage-gated Na+ channels are closed at rest. So, Na+ entry is negligible. The always operating Na+-K+
+
pump removes the Na from inside. This way the pump also creates negativity (a negative potential) inside
(removes 3Na+ in exchange of 2K+) and also maintain a high K+ inside which makes persistent K+ efflux
+ +
possible. If the Na -K pump is paralysed the RMP is lost. So, this pump is a very much important factor for the
maintenance of RMP. The RMP if disturbed by any means, various forces start acting immediately to restore the
+ -
MP. The most important mechanism is further K efflux (rectification current). Movement of Cl also occurs
but it is transient and insignificant. Even these forces are overwhelmed the MP is reversed and action potential
develops.

Action potential
Action potential (AP) is defined as the sequence of changes in the membrane potential in excitable cells due to
opening and closure of different ion channels after application of a threshold stimulus. So, action potentials are
the rapid changes in the membrane potential. In resting state the inside of the cell remains negative with
respect to the outside and this stage is called polarized state. If this polarized state in an excitable cell like
nerve or muscle is disturbed by a stimulus then the membrane potential changes in a specific manner due to the
movement of various ions leading to depolarization state (i.e. inside of the cell is positive). So, each action
potential begins with a sudden change from the normal negative resting membrane potential (RMP) to a
positive membrane potential and then ends with an almost equally rapid change back again to the negative
potential.

A typical action potential is seen when a nerve or a muscle fiber is stimulated by a threshold stimulus. The
stimulus can be defined as an agent that can change the polarized state of a cell with or without production of
action potential (AP). A stimulus may be physical, chemical, mechanical or electrical in nature. The stimulus
which is just sufficient to produce an action potential (i.e. to depolarize) in an excitable cell is called a
threshold stimulus to that cell. Stimuli stronger than this threshold stimulus is called suprathreshold stimuli
and the stimuli weaker than this threshold stimulus is called subthreshold stimuli. A subthreshold stimulus will
be unable to stimulate a tissue but a suprathreshold will of course stimulate. To be stimulated, a more excitable
tissue needs a less strong stimulus and a less excitable tissue, a stronger stimulus. But the true measure of
excitability is the chronaxie. The minimum strength which can stimulate a tissue is called rheobase. The time
required by rheobase to stimulate a tissue is called utilization time. A current weaker than a rheobase will
never stimulate a tissue. If a stimulus of double the strength of the rheobase is applied the time required to
stimulate the tissue is called chronaxie.
Stages of action potential
1. Resting state
2. Latent period
3. Depolarization stage
4. Rapid repolarization stage:
5. Negative after potential (after depolarization)
6. Positive after potential (after hyperpolarization/undershoot)
7. Resting state

Action potential in details:

The action potential starts at the site of application of the stimulus and moves along the membrane in all
directions. It shows a constant speed and amplitude under standard conditions in respect to ion concentration,
temperature, etc. A typical action potential is seen when a nerve or a muscle fiber is stimulated by a threshold
stimulus. When the stimulus is applied after dome delay (latent period) there is a gradual change of membrane
potential towards up to a certain point. This point is called firing potential as the membrane fires at this
membrane potential and then a sharp change is seen and membrane potential rapidly increases become positive
(+35mV) at this point the inside of the cell has become positive with respect to the outside. Now, the cell is said
to be depolarized. Then, the membrane potential changes in reverse direction i.e., towards the resting value or
RMP. At first it returns very quickly called rapid repolarization and then slowly called “negative after
potential” or “after-depolarization”. After this, the membrane potential goes below the original potential (i.e.
more negative than RMP), this stage is called “positive after potential” or “after-hyperpolarization”. Lastly
it comes back to RMP. The pointed peak plotted in a graph due to depolarization and rapid repolarization is
called spike. Normally action potentials are recorded as spikes and other features are not seen because the
events are very quick.

Ionic basis of action potential

This was discovered by Hodgkin and Huxley in 1950's. At rest, when the cell is polarized, the ECF contains
high sodium ions, whereas in ICF, it is low. But the potassium ion in the ICF is high and is low in the ECF.
Despite the difference in concentrations, free flow of sodium and potassium is impossible because of mainly,
non permeability of the cell membrane. There are channels in the cell membrane, called ion channels. These
ion channels are usually specific. Sodium channels admit only sodium ions and potassium channels admit only
+
potassium ions and so on. In resting state, both the concentration and electrical gradient of Na are directed
inwards and Na+ always wants to rush in. Whenever the Na+ channels are opened, the Na+ enters inside.
An action potential will not occur until the initial rise in membrane potential is great enough to create the threshold
level of the potential inside the cell membrane. After an application of the stimulus some time is required before any
appreciable change occurs in the membrane potential. This period is called latent period. The crossing of the
+ +
threshold level triggers the opening of voltage–gated Na channels in the membrane. Na now enters the cell and
+
reduces the membrane potential further. This lowering of membrane potential further opens more Na channels. It
+
continues in a positive feedback manner (i.e. more the number of Na ions enter the cell more will be the number
+
of the Na channels opening up) up to a point and the membrane potential reaches a critical value (firing potential).
+ +
Then, there is opening of maximum number of Na channels. It leads to explosive Na entry and the rapid upshoot
membrane potential and depolarization. The membrane potential is now positive and the cell is depolarized and
+
the cause of this depolarization is explosive Na entry. This stage of more positive ions inside the cells is referred to
as depolarization or activation stage. Depolarization requires 2-3 mili seconds and the time depends on ratio of
+ + +
Na outside and inside the cells, membrane permeability to Na and previous resting potential. Some K channels
+
are always open but with the start of decrease of membrane potential, plenty of voltage gated K channels
+
open up and K leaks to the outside rapidly. This is helped by positivity of the inside and the membrane potential
+
is rapidly restored. This results rapid repolarization and the K exit is called rectification current. After this, the
+
inside becomes negative and K efflux slows down resulting in “negative after potential” or “after-
depolarization”. It is believed that the negative after potential results partly from a buildup of potassium ions
immediately outside the membrane; this causes the concentration ratio of potassium across the membrane to be
temporarily less than normal and therefore prevents full return of the normal resting membrane potential for a
few additional milliseconds. The process continues due to late closure of the voltage-gated K + channels and
there is over correction of membrane potential; this causes “positive after potential” or after“-
hyperpolarization”. It is a fraction of a millivolt to a few millivolts more negative than the normal resting
membrane potential, but it can last from 50 millisecond to as long as many seconds. The first part of this
positive after potential is caused by the excess permeability of the nerve membrane to potassium ions at the end
of the spike potential. However, the prolonged continuation of this potential is caused principally by the
electrogenic pumping of excess sodium outward through the membrane. Later on, the membrane potential
comes to the original level.
+ + +
Ionic status is restored by Na -K pump during rest. This pump is stimulated by excess Na inside and
excess K+ outside. During one action potential a very small number of ions changes side (the cell loses some
K+ and gains some Na+) and in a cell many action potentials can form without rest.

K+ has tendency to go out and when the membrane potential is lowered the tendency to efflux increases which
+ +
is highest when the inside becomes positive. K goes out quickly in those above conditions, if further K
channels open.

Propagation of action potential

An action potential elicited at any one point on an excitable membrane usually excites adjacent portions of the
membrane resulting in propagation of the action potential. An excitable membrane has no single direction of
propagation. The action potential can travel in both directions away from the stimulus and even along all
branches of a nerve or muscle fiber until the entire membrane has become depolarized. A local circuit of
current flows between the depolarized areas of the membrane and the adjacent resting membrane areas. Positive
electrical charges flow inward through the depolarized membrane and then for several millimeters along the
core of the axon muscle fiber. These positive charges increase the voltage for a distance of 1-3 mm inside large
fibers to above threshold voltage value for initiating an action potential. Then, the sodium channels in these new
areas immediately activate and the explosive action potential spreads. And these newly depolarized areas cause
local circuits of current to flow still further along the membrane thus causing progressively more and more
depolarization. Thus, the depolarization process travels in both directions along the entire extent of the fiber.
The transmission along a nerve fiber and muscle fiber is called a nerve impulse and muscle impulse
respectively. Duration of action potential in nerve fiber is about 1 millisecond.

Plateau in some action potentials

In some cases the excitable membrane does not repolarize immediately after depolarization but instead the
potential remains on a plateau near the peak of the spike sometimes for many milliseconds before repolarization
begins. This type of action potential is found in cardiac muscle where the plateau lasts for as long as 3-4th tenths
second and causes contraction of cardiac muscle during the entire period of time.

Two separate types of channels are responsible for this type of action potential in cardiac muscle:
a. The usual voltage gated sodium channels
b. voltage gated calcium-sodium channels

The voltage gated sodium channels are also called the fast channels in case of cardiac muscles. The voltage
gated calcium-sodium channels are slow to be activated and therefore called slow channels. The voltage
gated calcium-sodium channels allow diffusion mainly of calcium ions and some sodium ions as well.
Activation of the fast channels causes the spike portion of the action potential whereas the slow but prolonged
activation of the slow channels is mainly responsible for the plateau portion of this type of action potential.
This type of action potential in case of cardiac muscles will be discussed in details while dealing with
cardiovascular system in further lectures.

All or none law

If a too weak stimulus is applied, there will be no depolarization. Once the strength of the stimulus is adequate
an action potential develops. However, a supra adequate strong stimulus will not produce stronger action
potential. At resting condition the cell membrane potential is at -90mv but it will cross 30mv at depolarized
condition. Thus, the potential will increase by 120mv. If the strength of the stimulus is increased, then also the
action potential will only cause the potential rise by 120mv, not more than that. It is the all or none law. Thus,
the stimuli crossing the threshold level is able to produce the action potential and however strong the stimuli be
the height of the potential will be same.

Refractory period
The membrane will not have new action potential unless the membrane is still depolarized from the preceding
action potential; this is called the refractory period. The interval between two successive action potentials in a
continuing stimulus is the refractory period of the cell membrane. The reason for this is that shortly after the
action potential is initiated the sodium channels become inactivated and any amount of excitatory signal applied
to these channels at this point will not open the inactivation gates. The only condition that will reopen them is
for the embrane potential to return either to or almost to the original resting membrane potential level. Then,
within another small fraction of a second, the inactivated gates of the sodium channels open and a new action
potential can then be initiated. The period of time during which a second action potential cannot be elicited,
even with a very strong stimulus, is called the absolute refractory period. This period for large myelinated nerve
fiber is about 1/2500 of a second.

Activation and inactivation of voltage gated sodium channels

The voltage gated sodium channels has two separate gates, one near outside of a channel called “activation
gate”and another near the inside called the “inactivation gate”. In the normal resting stage when the membrane
potential is -90 mV, the “activation gate”is closed that prevents any entry of sodium ions to the interior of the
fiber through these sodium channels. On the other hand, the “inactivation gate”is open and does not at this time
constitute any barrier to the movement of sodium ions.

Activation of the sodium channel: When the membrane potential becomes less negative than during the
resting state, rising from -90mv toward zero, it finally reaches a voltage, usually somewhere between -70and
-50 mc that causes a sudden conformational change in the “activation gate”,flipping it to open position. This is
called the activate state and during this state sodium ions can literally pour inward through the channel
increasing the sodium permeability of the membrane as much as 500- fold to 5000 fold.
Inactivation of the sodium channel: The same increase in voltage that opens the activation gate also closes the
inactivation gate. However, the closure of the “inactivation gate occurs as few 10,000ths of a second after the
activation gate opens; i.e. the conformation change that flips the “inactivation gate”to the closed state is a
slower process while the conformation change that opens the “activation gate”is a very rapid process.
Therefore, after the sodium channel has remained open for a few 10,000ths of a second, it suddenly closes and
sodium ions can no longer pour to the inside of the membrane. At this point the membrane potential begins to
recover back toward the resting membrane state which is the repolarization process. A very important
characteristic of the sodium channel inactivation process is that the inactivation process is that the inactivation
gate will not reopen again until the membrane potential returns either to or nearly to the original resting
membrane potential level. Therefore, it is not possible for the sodium channels to open again without the nerve
fiber first repolarizing.

Voltage gated potassium channels and their activation

During the resting st6ate, the gate of the potassium channel is closed and potassium ions are prevented from
passing to the exterior. When the membrane potential rises from -90mv toward zero. When the membrane
potential rises from -90mV toward zero this voltage change causes a slow conformational opening of the gate
and allows increased potassium diffusion outward through the membrane. However, because of the slowness of
the opening of these potassium channels, they open at the same time that the sodium channels are becoming
inactivated and therefore are closing. Thus, the decrease in sodium entry to the cell and simultaneous increase in
potassium exit from the cell greatly speeds the repolarization process, leading within a few 10,000ths of as
second to full recovery of the resting membrane potential.

MECHANISM OF MUSCLE CONTRACTION

General mechanism of muscle contraction
The sequence of events leading to contraction is initiated in the central nervous system, either as voluntary
activity from the brain or as reflex activity from the spinal cord. The somatic motor nerve which is going to
supply a skeletal muscle breaks up into several branches (axon terminal). Each axon terminal supplies one
muscle fiber. The end of the axon terminal is swollen like a bulb (terminal button or synaptic knob), which
fits into a depression of the muscle cell. The synaptic knobs contain acetylcholine (Ach) stored in clear
vesicles). The related part of the muscle membrane (sarcolemma) is thickened and is called motor end plate
which contains receptors for Ach. The motor end plate is folded to form the junctional folds. This folding
accommodates sufficient number of acetylcholine receptors for the transmission of the impulse. The space
inside the junction in between the nerve membrane and muscle membrane is called neuromuscular cleft or
synaptic cleft. When a motor neuron is activated, and an action potential passes outward in a ventral root of
the spinal cord. The action potential travels along the motor nerve up to its ending; there is calcium entry
into the synaptic knob. This is because of opening of voltage gated calcium channels by action potential.
This calcium causes exocytosis of the vesicles and release of acetylcholine into the neuromuscular cleft or
synaptic cleft. The acetylcholine molecules then cross the synaptic cleft within a millisecond and attach to the
acetylcholine receptors on the motor end plate. This combination leads to opening of channels in the centre of
each receptor through which sodium enters into the muscle cell. Therefore, opening of the acetylcholine
channel allows large quantities of sodium ions to flow to the interior of the muscle fiber at the point of the nerve
terminal. This causes the electrical resting potential under the motor end plate to change, and this then initiates
an action potential which passes in both directions along the surface of the muscle fiber. The action potential
depolarizes the muscle fiber membrane and also travels deeply within the muscle fiber through the T-tubules.
At each point where a transverse tubule touches part of the sarcoplasmic reticulum, it causes the sarcoplasmic
reticulum to release Ca++ ions. The calcium ions result in movement of troponin and tropomyosin and initiates
attractive forces between the actin and myosin filaments, causing them to slide together, which is the contractile
process. After a fraction of second, calcium ions are pumped back into the sarcoplasmic reticulum, where they
are stored till new action potential generates.

73
 Muscle contraction flow chart

Contraction Phase
Resting state

Motor nerve action potential arrives at motor end plate

Acetylcholine is released and sarcolemma and membranes are depolarized (Na+ flux into fiber) Action
potential is transmitted via T-tubules to SR
++ ++
Ca are released from SR terminal cisternae into sarcoplasm Ca are
bound by troponins

Myosin ATPase is activated and ATP is hydrolyzed Tropomyosin

shifts from actin binding site Actin-myosin cross-bridge

formation occurs

Repeated formation and breaking of cross-bridges resulting in sliding of filaments and sarcomere shortening
Relaxation Phase
Cholinesterase is released and acetylcholine is brokendown

Sarcolemma and T-tubules get repolarized

++ ++
SR Ca pump is activated and Ca ions return to SR terminal cisternae with the help of calsequestrin,
a calcium binding protein.

Actin-myosin cross-bridge formation terminates

Tropomyosin returns to actin binding site

Mg++ complex is formed with ATP

Passive sliding of filaments takes place

Sarcomeres return to the resting state

Excitation contraction coupling:

The process by which the excitation of the muscle is linked with the contraction or in other words how the
electrical phenomenon is converted into a mechanical one is called as excitation contraction coupling.

Molecular mechanism of muscle contraction - sliding theory

of muscle contraction
According to this theory, in relaxed state the ends of actin filaments derived from two Z-discs barely overlap
each other while at the same time completely overlapping the myosin filaments. But during the contracted state
these actin filaments are pulled inward among the myosin filaments so that they now overlap each other to a
major extent. Also the Z discs are pulled by actin filament up to the ends of the myosin filaments. So, muscle
contraction occurs by a sliding filament mechanism. But what causes the actin filaments to slide inward
among the myosin filaments? The mechanical, chemical or elelectrostatic forces generated by the interaction
of the cross bridges of the myosin filaments with the actin filaments must have caused the actin filaments to
slide inward among the myosin filaments. And most probably this is caused by the interaction of the cross-
bridges from the myosin filaments with the actin filaments. And indeed, the actin filaments can be pulled
together so tightly that the ends of the myosin filaments actually buckle during very intense contraction. Thus,
muscle contraction occurs by a sliding filament mechanism. Under resting conditions the sliding forces
generated by the actin and mysosin filaments are inhibited but when an action potential travels over the muscle
fiber membrane this causes the release of large quantities of the calcium ions into the sarcoplasm surrounding
the myofibrils. These calcium ions activate the forces between the filaments and contraction begins. The energy
required for the contractile process to proceed is derived from the high energy bonds of adenosine triphosphate
(ATP) which is degraded to adenosine diphosphate (ADP) to give the energy required.
Figure: Diagram showing the positions of actin and myosin filaments during the contracted and relaxed
state

Relaxed stage

Contracted stage
Interaction of myosin filaments, actin filaments and calcium to initiate contraction
A pure actin filament without the presence of troponin- tropomyosin complex binds strongly with myosin
molecule in presence of magnesium and ATP, which are in abundant number in myofibrils. If the troponin-
tropomyosin complex is added to the actin filament, the binding does not take place between the actin
and myosin filaments. It is because the active sites on the normal actin filament of the relaxed muscle are
inhibited or actually physically covered by the troponin-tropomyosin complex. In presence of large quantities
of calcium ions, the inhibitory effect of troponin-tropomyosin on the actin filament is itself inhibited.
When calcium ions combine with troponin C, each molecule of which can bind strongly with up to four
calcium ions and the troponin complex supposedly undergoes a conformational change that in some way
tugs on the tropomyosin molecule and moves it deeper into the groove between the two actin strands. This
uncovers the active sites of the actin, thus allowing contraction to proceed. The released calcium binds with
the troponin-C and thus the myosin head can bind with actin filament at its active sites.

Figure: Diagrammatic representation of actin filaments showing

F-actin, G-actin strands, tropomyosin strands, troponin-C, troponin-I and troponin-T.

Interaction between activated actin filament and myosin cross-bridges (Walk along theory of contraction)
When the actin filament becomes activated by calcium ions, the heads of the cross-bridges from the myosin
filament immediately becomes attracted to the active sites of the actin filament. When the head of the myosin
attaches with the active sites of the actin filament, there is conformational changes in head and the arm of the
cross bridges of the myosin molecules. This causes the head to tilt toward the arm and in doing so; it pulls the
actin filament towards it which is called the power stroke. Then, immediately after tilting the head
automatically breaks away from the active site and returns to its normal perpendicular direction. In this
position it combines with an active site farther down along the actin filament then a similar tilt takes
place again to cause a new power stroke and the actin filament moves another step. Thus, the heads of the
cross bridges bend back and forth and step by step walk along the actin filament, pulling the action toward the
center of the myosin filament. This hypothesis is called as the walk along theory of contraction. Each one of
the cross bridge operates independently of all others each attaching and pulling in a continuous but random
cycle. So, the greater the number of cross bridges in contact with the actin filament at any given time the
greater is the force of contraction. But immediately head leaves the site and returns back to get attached to a
new site. Thus, the myosin head moves from one active site to the next and the actin filament is pulled towards
the myosin filament therefore causing the muscle to contract.
Role of ATP as the source of energy for contraction
When a muscle contracts against a load the work is performed and for this energy is required. A large number
of ATPs are cleaved to form ADP during contraction process. And greater the amount of work performed by the
muscle the greater would be the amount of ATP that is cleaved. This is called the Fenn effect. Sequence of
events by which the ATP is used as a source of energy for muscle contraction is as follows:
a. Before the contraction begins, the heads of the cross-bridges bind with ATP.
b. The ATPase activity of the myosin head cleaves the ATP but leaves the cleavage products ADP and P i
(inorganic phosphate), bound to the head. In this state, the head of the myosin is perpendicular towards the
actin filament but is not yet attached to the actin.
c. When the inhibitory effect of the tropomyosin-troponin complex is itself inhibited by calcium ions, active
sites on actin filaments are made free and the myosin head bind with these.
d. This binding between the head of the cross bridge and the active site of the actin filament brings some
conformational changes in the head therefore causing the head to tilt backward toward the arm of the cross
bridge. This provides the power stroke for pulling the actin filament.
e. Once there is tilting, there is release of ADP and Pi which were previously attached to the head and this
exposes a site on the head where a new molecule of ATP binds. So, this binding of the new molecule of
ATP in turn causes detachment of the head from the actin.
f. After the head has split away from the actin the new molecule of ATP is also cleaved and the energy again
cocks the head back to its perpendicular condition ready to begin a new power stroke cycle.
g. When the cocked head with its stored energy derived from the cleaved ATP binds with a new active site on
the actin filament it becomes uncocked and once again provides the power stroke.
h. Similarly, the process proceeds again and again until the actin filament pulls the Z-line up against the ends
of the myosin filaments or until the load on the muscle becomes too great for further pulling to occur.

Energetics of muscle contraction

Muscle can be regarded as a machine that converts chemical energy into work. The first and second laws of
thermodynamics apply to muscle the same way as they apply to other machines. When a muscle contracts
against a load, it performs work. Thus, the energy is transferred from the muscle to the external work. The
muscle contraction is thus dependent upon the source of energy which is derived from ATP. ATP is required in
the functioning of calcium and Na+- K+ pump, which is essential in pumping the calcium to the sarcoplasmic
reticulum after the completion of the contraction and also in maintaining the level of sodium and potassium in
the fiber. The ATP present in the muscle is sufficient to maintain full contraction for only 1-2 seconds at
most, after the ATP is split to ADP, it is rephosphorylated into new ATP within a fraction of a second.
There are several sources of energy for this rephosphorylation. The first source of energy that is used to
reconstitute the ATP is the substance phosphorylcreatine, which carries a high energy phosphate bond similar
to that of ATP. The creatine is synthesized in liver from amino acids as methionine, glycine and arginine. In the
skeletal muscle the creatine is phosphorylated to phosphorylcreatine, which is stored in the muscle as a
high energy rich substance. The phosphorylcreatine is hydrolyzed to creatine and phosphate group to release
considerable amount of energy.

During rest condition, some ATP transfers its phosphate group from mitochondria to creatine and converts it to
phosphorylcreatine. During exercise, the phosphorylcreatine is hydrolyzed at the junction between the myosin
head and the actin and help in formation of ATP from ADP and help in continuation of contraction. The next
important source of energy used to reconstitute the ATP and phosphorylcreatine is glycogen previously stored
in the muscle cell. Rapid enzymatic breakdown of the glycogen to pyruvic acid and lactic acid liberates energy
that is used to convert ADP to ATP, and this ATP can then be used directly to energize muscular contraction or
to reform the stores of phosphorylcreatine.

ATP, Phosphoryl creatine and Glycogen Provide Energy for Muscle Contraction
The direct energy source for muscle contraction is ATP. The contractile protein, myosin, hydrolyzes ATP to
ADP + Pi, the used ATP is quickly restored. The depleted store of phosphorylcreatine is restored from ATP
when the muscle is returned to the resting state. ATP is synthesized via glycolysis in the sarcoplasm and/or via
oxidative phosphorylation in the mitochondria. Glycolysis may occur under anaerobic condition, the yield is 2
ATP/glucose. The lactate produced from glycogen is oxidized to yield 36 ATP/glucose.

Carbohydrate and lipid breakdown

At rest and during light exercise, muscles utilize lipids in the form of free fatty acids as energy source. As
intensity of exercise increases, lipid alone cannot supply energy fast enough, so utilization of carbohydrate
becomes predominant component in muscle fuel mixture. Thus during exercise, most of energy for
phosphorylcreatine and ATP resynthesis comes from breakdown of glucose to carbondioxide and water.
Types of muscle contraction
It is seen that contraction of muscles either may result into shortening or development of tension.
Accordingly the muscle contraction is of two types. They are as follows:

1. Isotonic contraction (isotonic implies same tension)

Muscle contraction is said to be isotonic when it does shorten with the tension on the muscle
remaining constant. Example: a person lifting a load. As the muscles shorten, the hand moves
and the load is lifted (i.e. displacement occurs). Force developed in the muscle (tension) in lifting
the load is constant throughout the movement. So, in this type of contraction external work is
done which is the measure of the developed tension in the muscle.

2. Isometric contraction (isometric implies same measure)

Muscle contraction is said to be isometric when the muscle does not shorten during contraction.
The muscle does not shorten but develops force (tension). Here the muscle contracts but its
length remains unchanged. Example: a person pulling stout rod. There is no movement of the
rod, i.e. no movement of the hands; it means the muscles though contracting severely cannot
shorten. Here, the force is generated in the myosin side chain due to power stroke but no
recycling of cross bridges occurs. In this type, therefore, no external work is done as there is
no displacement.

Rigor:
When muscle fibers are completely depleted of ATP and phosphoryl creatine, they develop a
state of rigidity called as rigor. In rigor almost all head attach to actin. After contraction, the
calcium should be pumped back into the longitudinal portion of the sarcoplamic reticulum. And
this is done by calcium pump called Ca-Mg ATPase. So, it is obvious that muscle relaxation
also requires ATP and is an active process. If the calcium cannot be removed from the cytosol
or there is no supply of ATP for detachment of myosin heads then the muscle remains in a state
of continued contraction. The condition is called contracture and this contraction occurs in
absence of any stimulation.

Rigor mortis:
Rigor mortis is a similar condition due to failure of relaxation after death. In this condition, the
muscles contracts and becomes rigid even without action potentials. This rigidity is due to the
loss of all the ATPs, which are required for the separation of the cross bridges from the actin
filaments during the relaxation phase. The pumping of calcium ions from the sarcoplasm into the
sarcoplasmic reticulum is an active process and since there are no ATPs available the calcium
ions could not be pumped out by calcium-magnesium pump and therefore the muscles remain in
contracted state. So, several hours after death, all the muscles of the body go into a state of
contraction called as rigor mortis. But if the death occurs after exhaustion (say after a long fight)
then ATPs are already reduced and rigor mortis develop quickly. So, rigor mortis takes different
time to set in different individuals. The rigor mortis is lost when the muscle protein is degraded
due to autolysis. This autolysis is caused by the enzymes (lysozymes-proteolytic enzymes)
released from lysosomes some hours after death. Rigor mortis occurs more rapidly at higher
temperature.

Fatigue:
Prolonged and strong contraction of muscle leads to the well known state of muscle fatigue.
Muscle fatigue is defined as a loss of work-output leading to a reduced performance of a
given task. Fatigue may result from deleterious alterations in the muscle itself and/or from
changes in the neural input to the muscle. During prolonged exercise, e.g. marathon-running,
depletion of muscle glycogen, decrease in blood glucose, dehydration, or increase in body
temperature contribute to fatigue. During intense muscular activity, e.g. short-distance running,
lactic acid is formed via anaerobic glycolysis. It thus decrease the pH of the muscle; this may
2+
inhibit metabolic processes, disturb excitation-contraction coupling, Ca fluxes, myosin
ATPase activity, and thereby decrease work output.

Chemical composition of muscle

Muscle is almost entirely protein and a dilute aqueous salt solution. A small part of lipid
found in the muscle originates from phospholipids and cholesterol found in membranes of
plasmalemma, the sarcotubular system and other membranous sub cellular organelles. The
+
muscle cell contains relatively high concentration of K and phosphate and relatively low
+ -
concentration of Na and Cl ion. DNA and RNA concentrations in muscle are lower than in
most other animal cells. The protein present in the muscle may be grouped into sarcoplasmic
protein, myofibrillar proteins and stroma proteins.

Table: Proximate composition of mammalian skeletal muscle

Constituents Percentage Comments
Water 55-78 Water content varies inversely with
lipid content.
Protein 15-23 Smooth muscle has a slightly lower
protein percentage.
Lipid 1-20 Varies widely depending on neutral
lipid content.
Carbohydrate 1-2 Mostly glycogen in rested muscle
and lactate in exhausted muscle
Ash 1
Nucleic acid <1
Other soluble organic compounds 1