EXCITABLE TISSUES:

These are tissues that are capable of eliciting response when acted upon by a stimulus.
Examples include muscles (skeletal, cardiac and smooth), nerves as well as glands.

SKELETAL MUSCLE
Structure of Skeletal Muscle fibers

Skeletal muscle is striated and is under voluntary control of the somatic motor system. The
skeletal muscle is an organ that consists of different tissues (blood vessels, the skeletal
muscle fiber, nerve fibers and connective tissue).

Skeletal muscle consists of three different connective tissues that enclose it, and
compartmentalizing each muscle fiber, thus providing it with its structural integrity and a
means by which it attaches to bones. Most skeletal muscles are attached to bones by collagen
fibers known as tendons or aponeurosis.

Each skeletal muscle is wrapped by a connective tissue called the epimysium. The
epimysium is an irregular sheath of connective tissue that separates the skeletal muscle from
other organs and tissues in the body thus allowing the muscle to contract maximally and
independently form other tissues while helping it retain its structural integrity. Inside the
Skeletal muscles are of bundles of muscle fibers called fascicles. The fascicule is organized
in such a way that only a group of fascicules within a skeletal muscle is activated by a group
of motor neurons thus eliciting specific movements. Each muscle fiber within the fascicule is
encased in a connective tissue called endomysium. The endomysium surrounds the muscle
cell and transfers the force of contraction of muscle fibers to the tendons attached to bones.
All three connective tissues intertwine to form the tendons.

All skeletal muscles are richly supplied with blood and have a rich innervation of the somatic
tree; each muscle fiber is innervated by an axon branch of the somatic motor neuron.

Skeletal muscle cells are long (can be as long as 27 cm in Sartorius muscle of the upper leg
and with a diameter of up to 100 µm) and cylindrical and are thus referred to as muscle
fibers. They are multinucleated, having large amounts of protein and enzymes needed for
maintaining normal cell functions. The skeletal muscle cell contains all basic cell organelles
like other cells, but some of the cell organelles are specialized to skeletal muscle cell having
quite different functions. The endoplasmic reticulum of skeletal muscle is known as the
sarcoplasmic reticulum and its function is to store, release and mop up calcium ions form the
sarcoplasm (muscle cell cytoplasm). The plasma membrane of the skeletal muscle fiber is
known as the sarcolemma which is a Greek word (sarco meaning flesh). In the sarcoplasm,
proteins are organized into structures known as myofibrils that run the length of the cell.

Microanatomy of the skeletal muscle

Skeletal muscle fibers (cells) consist of myofibrils (Figure 7.3) which are long protein
bundles of about 1µm in diameter. Each myofibril contains myofilaments known as actin and
myosin arranged in repeating units known as sarcomeres. The sarcomere is the functional unit
of the skeletal muscle

Figure 7.3: Skeletal muscle myofibril

cell. It is the shortening of sarcomeres that lead to contraction of individual skeletal muscle
fiber and ultimately the whole muscle.

7.4.1.1 The Sarcomere

A sarcomere consists of region of myofibril between two cytoskeletal structure called the Z-
discs or Z-line which gives the striated appearance of the skeletal muscle cell/fiber in addition
to the arrangement of the thin and thick filaments within each sarcomere. The dark A-band
consists of thick filament (myosin) that are located at the center of the sarcomere extending
towards the Z-line, myosin is static (does not move) and is anchored at the center of the
sarcomere by a protein called myomesin that makes up the M-line at the center of the
sarcomere. The lighter I-band consists of actin filaments attached to the Z-line by a protein
called α-actinin. The thin filaments extend into the A band and overlap the thick filament
(myosin) towards the M-line. The dark A-band is so because of the thick filament within this
region while the H-zone is located at the middle of the A-band and is slightly lighter in color
because the thin filaments do not extend towards this area. During muscle contraction the
filaments do not get shorter rather the thin filaments slide across the thick filament bringing
the Z-discs closer together thus shortening the muscle cell. The components of the thick and
thin filaments are discussed in `the following section.

Contractile Proteins - Actin, Myosin

Contractile proteins are proteins that mediate the sliding of contractile fibers or bring about
contraction in skeletal and cardiac muscles. There are two contractile proteins1, namely actin
and myosin.

Myosin: myosin `is ATP-dependent and is present in muscle as myosin-II (also known as
conventional myosin) - a very large protein of about 500kd and about 15nm in diameter.
Myosin consists of two pairs of light chains (called the essential and regulatory chains which
actively participate in muscle contraction) of about 20kd each and two heavy chains of 200kd
each consisting of about 2000 amino acids in length. The heavy chains have globular head
regions and long α-helical tails that coil around each other. In muscle cells the long, coiled
tails of individual myosin molecules join to form the thick filaments of the sarcomere. The
force producing heads stick out from the sides of the filament.

Actin: actin is a contractile protein found in eukaryotes with a mass of about 42kDa that form
microfilaments and are about 7nm in diameter. They form the thin filament in the contractile
unit of sarcomeres. The type of actin found on skeletal muscle is the F-actin. Actin is present
in both the A and I band, whereas myosin is only present in A band. Tropomyosin a
regulatory protein wraps around actin helix and covers its myosin binding site, at rest.

Mechanism of Muscle Contraction – The Sliding Filament theory

In 1954, three scientists: Andrew Huxley, Rolf Niedergerke and Hugh Huxley observed that
during contraction of the muscle, under high resolution microscopy, the “A band” composed
of myosin remained relatively constant in length, while the “I band” composed of actin
shortened (changed its length along with the sarcomere). These observations lead them to
postulate the “sliding filament theory”, which states that the sliding of actin past myosin
generates muscle tension. Since actin is attached to the lateral end of the sarcomere called “z
band” or “z discs”, any shortening of the actin filament over the myosin filament, would lead
to shortening of the sarcomere and ultimately, the muscle. The process by which actin and
myosin act to bring about muscle shortening/contraction has been termed “crossbridge
cycling”

Crossbridge cycling:

This is a series of molecular events that define the sliding filament theory, or better put, the
molecular basis of muscle contraction. The process by which depolarization of muscle fiber
initiates muscle contraction is known as excitation-contraction coupling.

At rest, troponin I is bound to tropomyosin and actin and covers the myosin binding sites
where myosin can bind with actin. At the same time, adenosine phosphate (ADP) is tightly
bound to the myosin head. Myosin-II molecules are dimers having two heads, but can only
attach to myosin, one head at a time. Following action potential leading to increase in cytosol
concentration of Ca2+, free Ca2+ ions bind to troponin C causing a conformational change and
weakening of troponin I’s interaction with actin. This, in turn, exposes myosin’s binding site
on actin. Once the binding site is exposed, myosin binds to actin, forming “myosin/actin
crossbridge”. Upon formation of the crossbridge, ADP attached to myosin head is released
causing a conformational change that moves actin thin filament medially along the thick
filament, known as the crossbridge “power stroke”. ATP further binds to the free site on the
myosin head leading to a detachment of the myosin head from actin. This process is
continued for as long as free calcium ions remains in the cytoplasm; with sufficient ATP, the
cycle repeats itself eliciting contraction. Each power stroke shortens the sarcomere by about
10nm. On each thick filament are about 500 myosin heads.

7.7 Regulatory Proteins -Troponin, Tropomyosin

Tropomyosin: Tropomyosin consists of two-stranded helical long filament proteins that line
the actin double helix. Each thin filament consists of about 300-400 actin molecules and 40-
60 tropomyosin molecules. On the tropomyosin lie troponin molecule which acts to regulate
actin myosin crossbridge by regulation of tropomyosin.

Troponin: troponin is a regulatory protein consisting of three subunits: troponin C, troponin

I and troponin T making the troponin complex that helps regulate muscle contraction.
Troponin is found in skeletal and cardiac muscle but not in smooth muscle. Troponin is
attached to tropomyosin which in turn wraps around actin. When a muscle is stimulated, free
calcium released from the sarcoplasmic reticulum attaches itself to the troponin C unit of
troponin causing changes that expose the myosin binding site on actin. Skeletal muscle
Troponin C has four calcium binding sites and acts to create a conformational change when
activated by calcium. Troponin I binds to actin in the thin myofilaments to hold the actin-
tropomyosin complex inhibiting the interaction of myosin and actin while, troponin T binds
to tropomyosin creating the troponin-tropomyosin complex.

7.8 Excitation-Contraction Coupling

Excitation contraction coupling was first termed and described by Alexander Sandow in 1952
and describes the fast physiological process that converts electrical stimulus to mechanical
response. It involves the rapid communication between electrical events occurring in the
sarcolemma of skeletal muscle fibers and Ca2+ release from the sarcoplasmic reticulum that
ultimately leads to muscle contraction. The sequence of events that lead to muscle contraction
involves the following:

1. Initiation of neural impulses (electrical activity): each muscle fiber is regulated/controlled

by a motor neuron, which conducts impulses from the brain or spinal cord. These
impulses are propagating action potentials that travel along the neuron to its axon
terminal. The terminal of the neuron connects to its muscle fiber at the neuromuscular
junction, where its consequent action potential leads to some physiological changes at the
neuromuscular junction.
2. Depolarization of the sarcolemma: Acetylcholine is the neurotransmitter released by the
neuron (from its motor endplate) into the neuromuscular junction in response to arrival of
the nerve impulse (action potential). Acetylcholine releases from the motor endplate into
the synaptic cleft binds to nicotinic receptors that change the resting membrane potential
of the muscle cell membrane (sarcolemma) by a Na+ dependent process. This generated
action potential on the sarcolemma propagates down and into the t-tubules that penetrate
the muscle fiber, thus, carrying electrical impulses deep into the muscle fiber. The T-
tubules help to carry impulses from the surface of the cell to the interior of the muscle
fiber to the dihydropyridine receptors (DHPR) facing the sarcoplasmic reticulum.
3. Interaction between dihydropyridine receptor and ryanodine receptor: Dihydropyridine
receptors are located along the T-tubules while the ryanodine receptor on the
sarcoplasmic reticulum. Action potential that spreads across the muscle fiber depolarizes
the inner portion of the muscle fiber thus activating the dihydropyridine receptors
(voltage-gated Ca2+ channel) which are in close proximity to the ryanodine receptors
(ligand-gated Ca2+ channel) on the sarcoplasmic reticulum. There is a physical interaction
between the ryanodine receptor and dihydropyridine receptor. Activation of the ryanodine
receptor initiate the release of Ca2+ from the sarcoplasmic reticulum into the cytoplasm.
4. Calcium induced calcium release: The release of calcium from the sarcoplasmic
reticulum induces further release of calcium into the cytoplasm. Ca+ released is ultimately
responsible for contraction of the muscle and its sustenance.
5. Muscle contraction: released Ca2+ binds to troponin which activates the myosin head and
initiates muscle contraction in a process known as the “sliding filament theory”.
6. Sequestration of Ca2+ into the sarcoplasmic reticulum: At the end of contraction,
intracellular Ca2+ concentration in the cytoplasm falls, as a result of its sequestration into
its intracellular storage site, the sarcoplasmic reticulum, by the action of Sarcoplasmic
reticulum Ca2+ ATPase (SERCA). The SERCA uses energy generated from ATP
hydrolysis to pump back the calcium in the cytoplasm back into the SR where it is stored
until its next release, upon activation.
7. Relaxation of the muscle fiber: Once the concentration of calcium in the cytoplasm is
sufficiently reduced, chemical interaction between the actin and myosin head ceases
leading to relaxation of the muscle fiber.

7.9 Characteristics of Skeletal Muscle –

7.9.1 Twitch, Motor Unit, Summation and Tetanus, Length-Tension Relationship

Twitch: One of the characteristics of skeletal muscles is the muscle twitch which is its ability
to contact rapidly from one neural stimulation as long as the neural stimulation is able to get
the muscle fiber form its resting membrane potential of about -80 to -90 mV to threshold
potential of about 45 mV.

The different phases of a Simple Muscle Twitch, showing: The Point of Stimulation (PS), the
onset of contraction (PC), the Contraction Phase (CP) and the Relaxation Phase (RP).
The muscle twitch consists of three phases: the latent, contraction and relaxation phases:

1. Latent Period: This is the period from application of a stimulus to the beginning of the
contraction (when calcium ions are being released)

2. Contraction Period: Here, the cross bridges are active, and the muscle shortens if the
tension is great enough to overcome the load.

3. Relaxation Period: Ca2+ ions are removed from the sarcoplasm by active transport of
Ca2+ back into the sarcoplasmic reticulum.

Summation and Tetanus:

If a muscle fibre is stimulated after complete recovery from this contraction, a second twitch
will be the same as the first twitch. If the muscle fibre is stimulated before it has completely
relaxed, the second twitch is added to the first twitch, resulting in summation. If a muscle
fibre is stimulated very rapidly, such that there is insufficient time for relaxation, a sub-
maximal, followed by maximal sustained contraction termed clonus (incomplete tetanus) or
tetanus occurs, respectively.

Skeletal muscle elicits graded contractions in response to demand for proper control of
voluntary movement. Muscle contraction can be graded in two ways: Increasing the
frequency of the stimulation and Increasing the number of motor units stimulated
(Recruitment, Figure 7.5).

Figure 7.5: A single stimulus results in a single contractile response – a muscle twitch (A) which consists of
contraction and relaxation. Additional frequent stimuli increase contractile force – wave summation (B); Further
stimulation results in a sustained contraction known as clonus (C, incomplete tetanus) or tetanus (D).

Length-Tension relationship:

The length-tension property of a whole muscle (or muscle fiber or sarcomere) is the
relationship between muscle length and the force the muscle can produce at that length. It is
an indication of the number of myosin heads actively involved in the crossbridge in the
sarcomere. When a sarcomere is at its maximum length, the number of myosin heads attached
to actin is fewer and as such, the tension is low; but as more myosin heads are activated and
pulled into the crossbridge the length of the sarcomere shortens and the amount of
force/tension within the sarcomere increases. The length of the sarcomere at which there is
maximum tension is between 1.6 to 2.6 µm, where most of the myosin heads are actively
involved in the contraction and is said to be the (optimum) resting length. (Figure 7.6). As
further shortening of the sarcomere continues fewer myosin heads are involved in the
contraction or shortening of the sarcomere because the actin strands/filament begin to overlap
over each other blocking the binding sites for myosin head and as such the tension in the
sarcomere reduces even though it is shorter, another reason for this is the absence of myosin
heads towards the center (M-line) of the sarcomere.