Department of Natural Sciences University of St.

La Salle Bacolod City

MUSCLE TISSUE- cells contain contractile proteins which generate the forces necessary for cellular contraction, and drives movement within organs and the body as a whole.  The cellular structure is organized in long units referred to as muscle fibers, which is an adaptation to its function of contraction.  The sarcoplasm appears fibrillar due to its contents of myofibrils. The staining is acidophilic or pinkish due to its affinity to acid stains.  The cells and fibers are bound together by varying amounts of areolar CT containing blood vessels and nerves. Their arrangement allows groups to act together or separately, generating contractile forces of varying strength.  Arises from mesodermal mesenchyme through accumulation of myofilaments & development of special membranous channels & compartments in the cytoplasm. Exception: iris smooth muscle cells are ectodermal.

Terminology associated with muscle tissue: 1. Sarcoplasm- acidophilic cytoplasm of muscle cells and fibers 2. Sarcolemma- cell membrane complex consisting of an external coating of protein polysaccharide over the sarcolemma. 3. Sarcosome- granules in the sarcoplasm which is actually a mitochondrion under EM. 4. Myofibrils- fine threadlike structures in the sarcoplasm which are responsible for contraction. Under EM, each myofibril is composed of finer myofilaments. 5. Sarcomere- a linear unit of muscle contraction 6. Sarcoplasmic reticulum- refers to the ER.

STRIATED or SKELETAL MUSCLE DISTRIBUTION: comprise most of the entire musculature of the body and are attached to the skeletal system.  Exceptions: tongue and the upper portion of the esophagus (striated, but involuntary in nature),  The contraction is quick, forceful and usually under voluntary control.

ORGANIZATIONAL LEVELS OF SKELETAL MUSCLE

HISTOLOGICAL VARIANTS 1.The skeletal muscle fiber is the biggest among the 3 types, and does not branch except in the muscles of the tongue. It has abundant areolar CT containing blood vessels intervening between sections of muscle fibers. 2.Fibers making up a single muscle are not uniform in cytological characteristics, becoming evident only in histochemical methods: a.Red muscle (slow fibers)- small dark fibers with relatively abundant sarcoplasm, fewer myofibrils, rich in mitochondria and myoglobin, possess a richer blood supply, thicker Z-lines, and complex profiles of sarcoplasmic reticulum; contraction is slow and steady; predominant in postural muscles and limbs.

b.White muscle (fast fibers)- paler fibers of large diameter, abundant myofibrils, granular sarcoplasm sparse, relatively narrow Z-lines, simple sarcoplasmic reticulum and sparse mitochondria. They react with quick, forceful contractions that cannot be sustained for long periods; predominate in extraocular muscles. c.Intermediate muscles- dispersed among red and white muscles where either type predominates.

 The histological criteria to differentiate skeletal muscle from other fibers are their numerous nuclei located peripherally just beneath the sarcolemma.  The sarcolemma has a protein (dystrophin) external coating with a delicate network of reticular fibers.  In Duchenne muscular dystrophy, this protein is lacking, resulting to structural weakness of the sarcolemma; damaged muscle fibers are replaced by CT and adipose tissue, that renders patients unable to walk by age 12.

 The sarcoplasm consists of a typical cytoplasmic matrix, the usual cell organelles and myofibrils characteristic of muscle tissue.  Due to the high energy demands of contraction, there are abundant, large mitochondria distributed in longitudinal rows between myofibrils.  Lipid droplets, glycogen (depot of energy mobilized during contraction), and the oxygen-binding protein myoglobin (necessary for the high oxidative phosphorylation level of the tissue) are inclusions of the sarcoplasm.  The interior is occupied by myofibrils, seen as fine dots uniformly distributed or grouped as polygonal shrinkage artifacts called Conheims area or field.  Bundles of myofibrils are arranged in characteristic Kollickers columns. They are so placed that the corresponding bands appear on the same transverse plane across the fiber.

 This arrangement results in the appearance of alternating dark (Aband) and light bands (I-band) on the fiber.  The I-band is dissected by a dark transverse Z-line or telophragma; the segment between 2 succeeding Z-lines is termed a sarcomere (includes an A-band and half I-bands on either side).  A paler zone, the Hband (also called intermediate disk of Hensen) traverses the center of the A band.

 In the center of this H-band is a narrow dark line, the Mline or mesophragma. The major protein of the M-line is creatine kinase which forms phosphocreatinine required for ATP synthesis.  EM reveals that myofibrils are composed of units termed myofilaments responsible for the observed crossstriations.

MYOFILAMENTS

Myosin resembles a golf club: its thin shaft is composed of the LMM (light meromyosin) chain; the heavier head chain (HMM) has actin and ATPbinding sites for contraction.

 In addition to actin and myosin, 2 other proteins are known to compose myofilaments: tropomyosin, and troponin.  Tropomyosin form filaments that run over the actin subunits alongside the outer edges of the groove between the 2 twisted actin strands. Troponin is a complex of 3 subunits: TnC binds calcium ions; TnT strongly attaches to tropomyosin; TnI inhibits actinmyosin interaction.

The spatial configuration of the 3 troponin subunits on F-actin hides the binding site of myosin heads to actin in the uncontracted muscle.

 Independent of the action of actin and myosin are other associated proteins (nebulin, tropomodulin, myomesin).  Titin is a third type of filament spanning the distance from the M-line of the A-band to the Z-disc.  They are responsible for myofibrillar passive elasticity, and maintain the central position of thick filaments in the sarcomere.

 The A-band is composed of thick myosin filaments held together in alignment by slender cross connections at the middle of the A-band, giving rise to the M-line.  The thinner actin filaments extend to either direction from the Z-line, constituting the I-band. They also extend some distance into the A-band.  The distance between their ends determine the length of the H-band which has no actin filaments (in the contracted state, the H-band is almost absent).

When a muscle contracts, the thin filaments slide past between the myosin filaments in the A-band

H-band is narrowed and the Z-line is drawn closer to the A-bands which shortens the myofibril.

 The sarcoplasmic reticulum is the sER of striated muscle cells, specialized to sequester Ca+2 ions.  It is a continuous system of membrane-limited sarcotubules extending throughout the sarcoplasm, forming a closemeshed canalicular network around each myofibril. It exhibits a repeating pattern of local differentiation in the sarcomere.  Longitudinal sarcotubules over the A and I junctions are confluent at regular intervals with pairs of transverse channels called terminal cisternae; pairs of these cisternae run transversely close to invaginations of the sarcolemma, the slender intermediate T-tubule.

 These 3 tubules form the triads of skeletal muscle. In mammalian muscle there are 2 triads per sarcomere, situated at the junctions of each A-band with I-bands.

 The sarcoplasmic reticulum and the triads of each Z-line are the submicroscopic structures involved in the inward spread of activation.  Two proteins have been isolated that are involved in signal transduction: an 1800kD protein binds ryanodine, a plant alkaloid that release calcium from the sarcoplasmic reticulum; a 212kD protein binds 1,4 dihydroxypyridine, a voltage dependent blocker of Ca+2 channels.

PHYSIOLOGY OF SKELETAL MUSCLE CONTRACTION

 When a muscle is stimulated, the sarcoplasmic reticulum releases calcium ions which changes the configuration of the actin-troponintropomyosin complex, exposing the myosinbinding site on actin.  The bending of the myosin head pulls the actin past the myosin filament.  The force responsible for this sliding filament theory of contraction is the breakdown of ATP by myosin ATPase localized in the cross bridges.  When ATP is not available, actin-myosin binding becomes stabilized (rigor mortis is the muscular rigidity that occurs shortly after death).

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INNERVATION
1.Motor (efferent)- the axons of motor neurons whose cell bodies lie in the spinal cord, branch and form myoneural junctions (motor-end plate) with several muscle fibers.  Motor unit consists of one motor neuron and all the muscle fibers it innervates. The number and size of motor units, as well as the control of the firing frequency of the activated units determine the intensity of a muscle contraction.  Motor-end plate appears as an arborization of the axon terminal closely apposed to the muscle fiber surface. In EM, each small axon lies in a groove on the surface of the muscle fiber showing a profuse folding of the sarcolemma.
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 The specialization beneath the axon terminal is termed the subneural apparatus.  Within the axon terminal are seen the synaptic vesicles (source of neurotransmitters, e.g. acetylcholine), and abundant mitochondria.  The depolarization at the motor-end plate is propagated into the fibers via the transverse tubule system.  At each triad, the depolarization causes the release of Ca+2, which initiates the contraction cycle.  When it ceases, calcium is actively transported back to the sarcoplasmic reticulum cisterna, TnC returns the TnI to a position in which it inhibits binding of the myosin head to actin, and the muscle relaxes.  Myasthenia gravis or muscular dystrophy is a progressive muscular weakness caused by a reduction in acetylcholine receptors in the myoneural junction.

2.Sensory (afferent)- the sensory end organ is a neuromuscular spindle lying in the perimysium, closely attached to a fasciculus, and running parallel to the surface.  It consists of a CT capsule within which lie striated intrafusal muscle fibers. The main afferent annulospiral nerve endings coil around these fibers. Intrafusal fibers, which function as stretch or proprioreceptors, also receive motor input from gamma afferents.

 Some muscles taper off at their extremities, where a myotendinous junction is formed. In this region, collagen fibers of the tendon insert themselves into complex infoldings of the plasmalemma of the muscle fibers.  The nerve endings supplying the junction are the encapsulated tendon organs or Golgi tendon organs which detect changes in muscle tension. An intense response of the tendon organ inhibits muscle contraction that can develop into potentially destructive excessive tension.

REGENERATION
 Skeletal muscle increases in volume (hypertrophy) by enlargement of existing fibers through an increase in the amount of sarcoplasm but not in the number of fibrils (hyperplasia); atrophy of disuse results to thinner fibers.  After destruction, myoblast-like stem cells form new myofibrils from normally quiescent satellite cells which are apposed with the surface of mature muscle fibers. Large defects are replaced by a CT scar.  For successful regeneration, connection with motor nerves is necessary.

CARDIAC MUSCLE DISTRIBUTION- found only in the myocardium of the heart, and in the walls of the large vessels joining the heart like the aorta, vena cava and the pulmonary vessels. Contraction is involuntary, vigorous and rhythmic.

HISTOLOGICAL CHARACTERISTICS
 In l.s., cardiac muscle fiber is long and branching, smaller than skeletal muscle, but also appears to be cross-striated.  The dark stripe (A or Q-band) is anisotropic; the light band (I or J-band) is clear and isotropic.  Each myofibril contains actin and myosin filaments.

 The myofibrils diverge around the single, centrally placed nucleus in the interior of the cell.  They are arranged in parallel bundles; a small amount of sarcoplasm separates each bundle.  Unlike skeletal muscle, there are lesser amounts of areolar CT, though richly supplied with small blood vessels and nerve plexuses.

 The fibers are not syncytial but are made up of separate cellular units joined by the intercalated discs, a unique characteristic of cardiac muscle. They occupy the Z-line of the I-band.  These discs are heavy, step-like transverse lines as seen in iron-hematoxylin stains. EM studies show that they are junctional complexes (desmosomes, zonula adherens/ zonula occludens, gap junctions) of apposing plasmalemma.

 They are anchoring sites for actin filaments and bind cardiac cells together to prevent their separation during contraction cycles.  Gap junctions permit a rapid spread of excitation from cell to cell, hence cardiac muscle appear to function as a syncytium.

 The sarcoplasm has interstitial granules or granules of Kollicker seen under LM as muscle granules of glycogen, fat, albumin, lipofuscin, pigment and mitochondria.  The latter contain closely-packed cristae with a zig-zag appearance. It is richer in glycogen than that of skeletal muscle.  The sarcoplasmic reticulum is not as highly developed but more numerous and larger than those in skeletal muscle.  Instead of triads, these simple plexiform arrangement of tubular elements with no terminal cisternae are composed only of dyads (T-tubule and 1 sarcoplasmic reticulum).  Aside from their role of coupling excitation to contraction, these channels are additional surfaces for exchange of metabolites between muscle and extracellular space.

 The TATS (transverseaxial-tubular) system occur only over the Z-lines of the ends of sarcomeres.  They wander irregularly through the myofilaments, hence discrete bundles are not present.

 Despite differences in structure, the composition and arrangement of myofilaments are similar in skeletal and cardiac muscle; hence their contraction are essentially the same at the cellular level.  Atrial muscle has fewer T-tubules than ventricular muscles.  Membrane-limited granules are found in the atria, containing precursors of cardiodilatins (CDD) or atrial natriuretic polypeptides (auriculin or atriopeptin).  This hormone causes vasodilatation, lowering blood pressure EM of an atrial muscle and decreased blood volume; cell showing the presence of natriuretic opposes the sodium and water granules aggregated at conservation actions of the nuclear pole. aldosterone and ADH.

SPECIALIZED CONDUCTING TISSUE OF THE HEART

Lesions in the following may cause asychrony in the timing of atrial & ventricular contraction resulting in impaired efficiency of the heart. 1.Sinoatrial node (node of Keith and Flack or pacemaker)- located beneath the epicardium at the junction of the superior vena cava and the right atrium. These are striated, slender, fusiform fibers that are continuous with atrial muscle fibers.

2.Internodal tractsconnect the sinoatrial and atrioventricular nodes 3.Atrioventricular nodesituated in the lower part of the interatrial septum

A merry heart doeth good like a medicine.

4.Atrioventricular bundle (neuromuscular bundle or bundle of His)located beneath the pericardium and composed of muscular elements called Purkinje fibers.  They penetrate the ventricular myocardium and come in contact with ordinary muscle fibers at their ends, where they appear to lose their specific cytological features.

 They are bigger, shorter, less branched with fewer nuclei, myofibrils, stripes, and a relatively greater amount of sarcoplasm which has abundant glycogen and mitochondria.  Few intercalated discs are found, but there are numerous desmosomes and gap junctions.

INNERVATION- heartbeat is myogenic, but modulated by ANS. Heart rate is slowed by the parasympathetic vagus nerve, and accelerated by sympathetic nerve stimulation. REGENERATION- cardiac muscle is more resistant to injuries, but shows very little evidence of regeneration after injury.  The coronary arteries supplying blood to the heart are anatomical end arteries and lack collaterals.  In the event of blockage of coronary arteries (as a result of a blood clot or atherosclerotic blockage), the cardiac myocytes vascularized by the coronaries cannot receive essential oxygen and the result is infarct.  Following infarcts, the remaining heart muscle undergoes compensatory hypertrophy, with subsequent enlargement of the heart.  Hypertrophied hearts are commonly an indication of underlying pathological disorders, though they may develop in specific cases of training and overload as in athletes.  Stem cell technology holds promise for damaged hearts.

SMOOTH MUSCLE
DISTRIBUTION:  Forms the contractile portion of the walls of the digestive tract from the middle of the esophagus to the internal sphincter of the anus.  Walls of ducts in glands associated with the alimentary canal.  Walls of the respiratory passages from the trachea to the alveolar ducts; in the urinary and genital ducts.  Walls of the arteries, veins and larger lymphatic vessels.  Capsules of certain organs like the spleen.  Arrectores pilorum muscles in the skin responsible for the so-called goose flesh  Iris and ciliary body of the eyes concerned with accommodation and with constriction and dilation of the pupil.  Areola of the mammary gland and in the subcutaneous tissue of the scrotum.

HISTOLOGICAL CHARACTERISTICS:  Smooth muscle fibers form smaller fascicles with meager perimysium. They are classified according to developmental, biochemical and functional differences: 1.Visceral smooth muscle- derived from splanchnopleural mesenchyme and found in walls of thoracic, abdominal and pelvic organs of the respiratory digestive and urogenital systems; contains actin, myosin and desmin; poor nerve supply; unitary muscle exhibits autorhythmicity via gap junctions; contraction is slow and in waves. 2.Vascular smooth muscle- differentiate in situ from mesenchyme around blood vessels; contains desmin and vimentin; also unitary muscle; contraction are not sustained and localized. 3.Smooth (sphincter and dilator) muscles of the iris- derived from ectoderm and has a rich nerve supply; multiunit muscle because cells can contract individually; capable of precise and graded contractions.

 Appears in l.s. as fusiform or spindle shaped cells with fine tapering ends and a wider central region in which the oval nucleus is situated.  The surface of each cell is invested by a thick extracellular coating which corresponds to the basal lamina of epithelia and containing collagenous, elastic and reticular fibers.  This is responsible for its positive response to the PAS reaction, and favors the transmission of the pull of contraction to neighboring cells since reticular fibers continue into surrounding tissue.

 In x.s. smooth muscle appear like a mosaic of rounded or irregularly polygonal structures of varying diameters. Only the biggest fibers show the nucleus.  A scanty amount of areolar CT containing blood vessels is seen between units; it is not as vascular as either skeletal or cardiac muscle.  Irregular dark staining areas traversing muscle cells are called contraction nodes/ bands believed to be the result of movement of electrolytes and granules within the muscle cell during contraction.

 The plasmalemma between sites of attachment of myofilament has gap junctions or nexuses, and vesicular pinocytic inpocketings similar to those found in endothelial cells.  After maceration in nitric acid, myofibrils appear arranged parallel to the long axis of the muscle cell.  These contractile elements are double refractile under the polarizing microscope, but do not show the alternating isotropic and anisotropic birefringent) transverse bands of striated muscle fibers.  There are slender mitochondria, scanty ribosomes and rER in the sarcoplasm.

 Thin, parallel myofilaments containing actin and myosin are associated in bundles, and overlap much more than striated muscle. This accounts for lack of striations, and permits greater contraction.  Myosin have heads along most of their length, are less stable in the cytoplasm, and interact with actin only when the HMM is phosphorylated in response to a contractile stimulus.  Since the tropomyosin complex of skeletal muscle is absent, Ca+2 complexes with the calcium-binding protein calmodulin, also involved in the contraction of non-muscle cells.  A rudimentary sarcoplasmic reticulum is present; Ttubules, dyads/triads are absent. Membrane-limited vesicles called caveolae aid in Ca+2 uptake and release.

Action of hormones, neurotransmitters (ACh) or deformation of the epithelial cell by flow of blood can trigger reactions which can stimulate or inhibit associated smooth muscle beds. These effects may operate via 2nd messenger systems such as phospholipase A2 (PLA2) which may in turn activate cyclo-oxygenase (COX) / prostacyclin synthase (PCS) enzyme systems to produce prostaglandins (PGI2) which diffuse readily through the tissue fluids to act on the smooth muscle cells. Activation of nitric oxide synthase (Larginase) (NOS) may result in production of gaseous "neurotransmitter" NO. These paracrine agents act on the smooth muscle cells either through G-protein systems or directly on ion channels.

 Abundant in smooth muscle are the intermediate filaments desmin (skeletin) and vimentin.  Two dense bodies that contain E-actinin of Z-lines of striated muscle are also present. They transmit contractile force to adjacent cells.

 The force of slow, wave-like contraction is transmitted through their investment of reticular fibers to the interstitial CT. It is under involuntary control from the ANS: a sympathetic innervation from the thoracic autonomic nerves and a parasympathetic innervation from the cranial and pelvic autonomic nerves.  Elaborate neuromuscular junctions are not present.  Autonomic nerve axons terminate in a series of dilatations in endomysial CT.  Smooth muscles maintain a state of partial contraction called tonus, subject to modulation by blood-borne hormones: norepinephrine, angiotensin and vasopressin, resulting in vasoconstriction; circulating estrogens also affect uterine smooth muscles.

VARIATIONS OF SMOOTH MUSCLE CELLS

 Pericytes found on surface of capillaries and post-capilary venules are assumed to be capable of contraction and influence blood flow.  Myoepithelial cells are smooth muscle cells that lie between the glandular epithelial cells and the basal lamina of glandular acini of salivary, sweat, lacrimal or mammary glands (also referred to as basket/basal cells due to their branching, stellate shape, or their location at the base of alveolus). They expel secretion from the acini to the ducts of the glands by contraction.

REGENERATION
 It may divide by mitosis, but this capacity is limited.  Defects in smooth muscle heal by scar formation developed from perivascular mesenchymal cells.  In the uterus, cells are capable of hyperplasia in the event of pregnancy. After delivery, hypertrophied cells return to their normal size, with the excess number undergoing fatty degeneration.  An abnormal proliferation of smooth muscle cells and their excess collagen contribute to artherosclerotic plaques in aging aorta and coronary arteries.

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