Histology SSN October 25, 2004 Muscle Nadia Goodwin – ng2021 Rebecca Nardi – ran2101 SKELETAL MUSCLE I. Organization A. General Muscle fibers run parallel and straight Muscle tissue has many fasciculi A fasciculus has many myofibers A myofiber has many myofibrils Myofiber = muscle cell “Muscle fiber” is really a syncytium of small myoblasts that fused to form a large multinucleated cell Polygon-shaped in cross section nuclei pushed to outer edge of cell by the myofibrils A myofibril is composed of many myofilaments (thick and thin) B. Surrounding connective tissue 1) Endomysium Loose connective tissue that surrounds each muscle fiber Reticular fibers Contains the smallest capillaries and neuronal branches 2) Perimysium Connective tissue + fat + blood vessels and nerves Group fibers into fascicles 3) Epimysium Dense connective tissue that surrounds muscle groups. Large vessels and nerves penetrate this to reach muscle fibers II. Myofilaments & Striations A. Types: 1) Thick: myosin 2) Thin: actin B. Sarcomere = Basic Contractile Unit C. Striations: Diagramed in sarcomere. Seen in entire muscle b/c myofibrils and myofibers are in register. H-band = myosin not overlapping actin A-band = all myosin (both overlapping and non-) I-band = actin not overlapping myosin Z disc = disc in center of I band from which actin extends 1 Histology SSN October 25, 2004 III. Function Tidbits A. Musculotendinous junction = to dense regularly-arranged connective tissue (tendon) i.e. collagen type I. B. Triad = functions to deliver intracellular Ca++ to initiate contraction. (2 triads/sarcomere) 1. (1) Transverse (T) Tubule = in-pocketing of plasma membrane 2. (2) cisternae of the Sarcoplasmic reticulum C. Motor unit = neuron + muscle cells it innervates. CARDIAC MUSCLE I. Organization Fiber = one, mononuclear cell Central nucleus (can be seen in cross section too) Columns of cells are branched II. Intercalated discs A. Gap junctions Lateral portion Important for ionic continuity, to coordinate contraction B. Fasciae adherens Transverse portion Holds cells together at their ends C. Desmosomes Lateral and transverse portions Keeps cells together ** Tight Junction = Zona occludans is not found in cardiac muscle III. Functional A. intrinsic rhythm - does not need ACh or norepinephrine for contraction 2 Histology SSN October 25, 2004 SMOOTH MUSCLE I. Characteristics Surrounds the walls of ducts, blood vessels, GI tract, etc. Function to give an imprecise squeeze (staggered sheets of cells) Nuclei cigar-shaped lie in the middle of the fiber appear wrinkled or “puckered” Plane of section determines visibility of nuclei, cell width No striations because of diffuse contractile elements Gap junctions allow for united contractions Basal Lamina (Collagen IV) and Reticular Fibers (Collagen III) surround individual fibers II. Smooth muscle vs. Dense irregularly-arranged connective tissue A. Color Smooth muscle is more purple because of combination of actin/myosin and RNA (red + blue = purple) Connection tissue is more red because of purity of collagen B. Nuclei Smooth muscle nuclei are inside cell Fibroblast nuclei are in between collagen fibers C. Homogeneity Smooth muscle is composed only of muscle Connective tissue contains fibroblasts, macrophages, lymphocytes, etc., and will therefore have a ragged appearance Skeletal Muscle Cardiac Muscle Smooth Muscle Striated Yes Yes No T Tubule Location A-I junction Z disk Sarcoplasmic Reticulum: 2 cisternae in Triad w/ T tubule No Anastamosing network Yes Yes Yes (limited) No Yes Level of Ca2+ regulation thin filaments thin filaments thick filaments Location of Nucleus Peripheral Central Central Electrically coupled via Gap Junctions? Can regenerate? 3 Histology SSN October 25, 2004 MUSCLE QUESTIONS 1) Which of the following statements is FALSE? a. The band marked “A” does not include actin filaments. b. The M line is found in the band marked “A”. c. During contraction, the length of the band marked “A” remains the same. d. During contraction, the length of the band marked “B” is shortened. 2) Which of these statements is FALSE about the muscle type at the pointer? a. The cells in this muscle type are electrically coupled. b. This muscle type utilizes a T-tubule system for calcium delivery. c. This muscle type is not capable of regeneration. d. Calcium regulation occurs at the level of thick filaments. 3) Which figure(s) demonstrates smooth muscle? a. Figure A&C b. Figure A&D c. Figure B&C d. Figure B&D 4) The structure at the pointer is composed of all of the following except: a. desmosomes b. tight junctions c. gap junctions d. fascia adherens 5) The structure at the pointer is a: a. site of calcium-binding protein (calsequestrin) and a Ca2+-activated-ATPase. b. site of glycogen storage and degradation to glucose c. site from while Ca2+ is passively transported during muscle relaxation d. site into which calcium is released during muscle relaxation e. form of rough endoplasmic reticulum 4 Histology SSN October 25, 2004 MUSCLE ANSWERS 1) a. This is skeletal muscle. A marks the A band and B marks the I band. The A band consists of myosin and some actin that overlaps the myosin. Thick filaments of myosin anchor at the M line, which is found in the middle of the H zone. The figure below demonstrates how the length of the A band stays the same during contraction, while the I band gets shorter. 2) d. This is cardiac muscle. The cells are electronically coupled via gap junctions, thus allowing contractile signals to pass from cell to cell and allowing cardiac muscle to behave as a syncytium. T tubules are used for calcium delivery in both skeletal and cardiac muscle. Smooth muscle cells have no T tubule system. Cardiac muscle is not capable of regeneration (mature cardiac muscle cells do not divide). If injury to cardiac muscle tissue leads to death of cells, fibrous connective tissue forms (scarring) with consequent loss of cardiac function. Since tropomyosin and troponin wind around the actin filaments, calcium regulation does not occur at the level of the thick filament. 3) b. A is smooth muscle and B is skeletal muscle (from the esophagus). C is an EM of skeletal muscle and D is an EM of smooth muscle. Skeletal muscle has striations whereas smooth muscle does not. The EM in Figure C shows a higher magnification of the H band, A band, I band, and Z line in a striated muscle fiber. Figure C also shows a triad of T tubules and flanking terminal cisternae, which is only found in skeletal muscle. Whereas skeletal muscle has multiple, peripherally-located nuclei, smooth muscle has a single, centrally located nucleus, as seen in Figure D. 4) b. Pointer is at an intercalated disc. An intercalated disc does not have tight junctions. The desmosomes are on both the transverse and lateral components of the intercalated disc, while the fasciae adherens are on the transverse. Note 5 Histology SSN October 25, 2004 from Dr. Rothman: Gap junctions can also be a component of the transverse portion of the disc but they are not as prevalent as on the lateral. 5) a. This is an EM of skeletal muscle. The pointer is at the sarcoplasmic reticulum. The sarcoplasmic reticulum (SR) provides a mechanism for the muscle cell to regulate the concentration of cytosolic calcium. It is a modified smooth endoplasmic reticulum that serves alternatively as a storage site and a source of cellular calcium. Calcium is actively transported from the cytosol to the SR through the activity of a Ca2+-dependent ATPase. Calsequestrin is a calciumbinding protein found in the SR that, as it’s name implies, functions in the sequestion of calcium. Calcium is released from the SR during muscle contraction and stored during relaxation. Glycogen is stored as particles or droplets in the cytoplasm, which contains the enzymes required for the synthesis and breakdown of glycogen. 6