scholarly journals The subcellular distribution of dystrophin in mouse skeletal, cardiac, and smooth muscle.

1991 ◽  
Vol 115 (2) ◽  
pp. 411-421 ◽  
Author(s):  
T J Byers ◽  
L M Kunkel ◽  
S C Watkins
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Plasma Membrane ◽  
Cardiac Muscle ◽  
Striated Muscle ◽  
Adherens Junctions ◽  
Myotendinous Junction ◽  
Elevated Concentration ◽  
Western Blots ◽  
Functional Roles

We use a highly specific and sensitive antibody to further characterize the distribution of dystrophin in skeletal, cardiac, and smooth muscle. No evidence for localization other than at the cell surface is apparent in skeletal muscle and no 427-kD dystrophin labeling was detected in sciatic nerve. An elevated concentration of dystrophin appears at the myotendinous junction and the neuromuscular junction, labeling in the latter being more intense specifically in the troughs of the synaptic folds. In cardiac muscle the distribution of dystrophin is limited to the surface plasma membrane but is notably absent from the membrane that overlays adherens junctions of the intercalated disks. In smooth muscle, the plasma membrane labeling is considerably less abundant than in cardiac or skeletal muscle and is found in areas of membrane underlain by membranous vesicles. As in cardiac muscle, smooth muscle dystrophin seems to be excluded from membrane above densities that mark adherens junctions. Dystrophin appears as a doublet on Western blots of skeletal and cardiac muscle, and as a single band of lower abundance in smooth muscle that corresponds most closely in molecular weight to the upper band of the striated muscle doublet. The lower band of the doublet in striated muscle appears to lack a portion of the carboxyl terminus and may represent a dystrophin isoform. Isoform differences and the presence of dystrophin on different specialized membrane surfaces imply multiple functional roles for the dystrophin protein.

scholarly journals Chemical and immunochemical characteristics of tropomyosins from striated and smooth muscle

1974 ◽  
Vol 141 (1) ◽  
pp. 43-49 ◽  
Author(s):  
Peter Cummins ◽  
S. Victor Perry
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Cardiac Muscle ◽  
Skeletal Muscles ◽  
Striated Muscle ◽  
Β Subunit ◽  
Muscle Type ◽  
Amphibian Species ◽  
Species Specific ◽  
Immunochemical Characteristics

1. On electrophoresis in dissociating conditions the tropomyosins isolated from skeletal muscles of mammalian, avian and amphibian species migrated as two components. These were comparable with the α and β subunits of tropomyosin present in rabbit skeletal muscle. 2. The α and β components of all skeletal-muscle tropomyosins contained 1 and 2 residues of cysteine per 34000g respectively. 3. The ratio of the amounts of α and β subunit present in skeletal muscle tropomyosins was characteristic for the muscle type. Muscle consisting of slow red fibres contained a greater proportion of β-tropomyosin than muscles consisting predominantly of white fast fibres. 4. Mammalian and avian cardiac muscle tropomyosins consisted of α-tropomyosin only. 5. Mammalian and avian smooth-muscle tropomyosins differed both chemically and immunologically from striated-muscle tropomyosins. 6. Antibody raised against rabbit skeletal α-tropomyosin was species non-specific, reacting with all other striated muscle α-tropomyosin subunits tested. 7. Antibody raised against rabbit skeletal β-tropomyosin subunit was species-specific.

Ultrastructural Evidence for Myosin of the Smooth Muscle Type at the Surface of Trypsin-Dissociated Embryonic Chick Cells

1974 ◽  
Vol 15 (2) ◽  
pp. 279-289
Author(s):  
I. AP GWYNN ◽  
R. B. KEMP ◽  
B. M. JONES ◽  
U. GRÖSCHEL-STEWART
Keyword(s):  
Smooth Muscle ◽  
Plasma Membrane ◽  
Striated Muscle ◽  
Smooth Muscle Myosin ◽  
Rabbit Serum ◽  
Embryonic Chick ◽  
Antibody Preparation ◽  
Rabbit Igg ◽  
Antibody Conjugate ◽  
Muscle Myosin

Cells dissociated from embryonic chick muscle tissue using trypsin were rotated in the presence of globulin-enriched rabbit antisera against both smooth and striated muscle actomyosins originating from chicken gizzard (GAM) and pectoralis (PAM) muscles respectively. The presence of the rabbit antibodies was demonstrated using peroxidase-labelled sheep anti-rabbit λ-globulins, the enzyme-antibody conjugate being located by electron-microscope histochemistry. Anti-GAM λ-globulins reacted strongly with the plasma membrane. Judging from the complete absence of staining, λ-globulins from non-immunized rabbit serum did not interact with the membrane.When λ-globulins of sheep anti-rabbit IgG serum were applied alone, that is in the absence of pretreatment with rabbit λ-globulin, there was an observable reaction with the cell surface. Preincubation of anti-GAM with the heavy meromyosin fraction from smooth-muscle myosin inhibited the interaction of the antibodies with the membrane, as evidenced by the absence of staining. A weak positive reaction obtained with anti-PAM was due to components of the antibody preparation which were reactive with actin and not with PAM. It was concluded that a smooth-muscle myosin-like protein is an integral part of the plasma membrane of embryonic chick muscle cells.

scholarly journals Immunohistochemical localization in normal tissues of different epitopes in the multidrug transport protein P170: evidence for localization in brain capillaries and crossreactivity of one antibody with a muscle protein.

1989 ◽  
Vol 37 (2) ◽  
pp. 159-164 ◽  
Author(s):  
F Thiebaut ◽  
T Tsuruo ◽  
H Hamada ◽  
M M Gottesman ◽  
I Pastan ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Plasma Membrane ◽  
Cardiac Muscle ◽  
Muscle Fibers ◽  
Transport Protein ◽  
Rat Tissues ◽  
Human Tissues ◽  
Normal Tissues ◽  
Skeletal Muscle Fibers ◽  
Muscle Myosin

Using peroxidase immunohistochemistry, we examined the distribution of P170, a multidrug transport protein, in normal tissues by use of two different monoclonal antibodies (MAb). MAb MRK16 is a MAb that has been shown to react with an epitope in P170 located on the external face of the plasma membrane of multidrug-resistant human cells. MAb C219 has been shown to react with P170 in many mammalian species, and detects an epitope located on the cytoplasmic face of the plasma membrane. Using MRK16, we have previously described the localization of P170 on the bile canalicular face of hepatocytes, the apical surface of proximal tubular cells in kidney, and the surface epithelium in the lower GI tract in normal human tissues. In this work, we report that MRK16 also detects P170 in the capillaries of some human brain samples. A similar pattern was found using MAb C219 in rat tissues. in addition, MAb C219 showed intense localization in selected skeletal muscle fibers and all cardiac muscle fibers in rat and human tissues. ATPase cytochemistry showed that these reactive skeletal muscle fibers were of the type I (slow-twitch) class. Other additional sites of C219 reactivity in rat tissues were found in pancreatic acini, seminal vesicle, and testis. Electrophoretic gel immunoblotting showed two protein bands reactive with MAb C219. In liver, MAb C219 reacted with a approximately 170 KD band. In skeletal and cardiac muscle, MAb C219 reacted with a approximately 200 KD band which migrated in the same position as myosin. This band also reacted with an antibody to skeletal muscle myosin. This result suggests that C219 may crossreact with the heavy chain of muscle myosin in cardiac and skeletal muscle. Because MAb C219 reacts with proteins other than P170, it should be used with caution in studies of multidrug resistance.

Cardiac and skeletal muscle mitochondria have a monocarboxylate transporter MCT1

1999 ◽  
Vol 87 (5) ◽  
pp. 1713-1718 ◽  
Author(s):  
George A. Brooks ◽  
Marcia A. Brown ◽  
C. E. Butz ◽  
James P. Sicurello ◽  
Hervé Dubouchaud
Keyword(s):  
Skeletal Muscle ◽  
Striated Muscle ◽  
Lactic Dehydrogenase ◽  
Monocarboxylate Transporter ◽  
Lactate Oxidation ◽  
Western Blots ◽  
Lactate Shuttle ◽  
Muscle Mitochondria ◽  
Monocarboxylate Transporter 1 ◽  
Skeletal Muscle Mitochondria

To evaluate the potential role of monocarboxylate transporter-1 (MCT1) in tissue lactate oxidation, isolated rat subsarcolemmal and interfibrillar cardiac and skeletal muscle mitochondria were probed with an antibody to MCT1. Western blots indicated presence of MCT1 in sarcolemmal membranes and in subsarcolemmal and interfibrillar mitochondria. Minimal cross-contamination of mitochondria by cell membrane fragments was verified by probing for the sarcolemmal protein GLUT-1. In agreement, immunolabeling and electron microscopy showed mitochondrial MCT1 in situ. Along with lactic dehydrogenase, the presence of MCT1 in striated muscle mitochondria permits mitochondrial lactate oxidation and facilitates function of the “intracellular lactate shuttle.”

scholarly journals THE ULTRASTRUCTURE OF FROG VENTRICULAR CARDIAC MUSCLE AND ITS RELATIONSHIP TO MECHANISMS OF EXCITATION-CONTRACTION COUPLING

1968 ◽  
Vol 38 (1) ◽  
pp. 99-114 ◽  
Author(s):  
Nancy A. Staley ◽  
Ellis S. Benson
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Muscle ◽  
Striated Muscle ◽  
Structural Features ◽  
Mammalian Skeletal Muscle ◽  
Striated Muscles ◽  
Thin Filaments ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Dense Granules

Frog ventricular cardiac muscle has structural features which set it apart from frog and mammalian skeletal muscle and mammalian cardiac muscle. In describing these differences, our attention focused chiefly on the distribution of cellular membranes. Abundant inter cellular clefts, the absence of tranverse tubules, and the paucity of sarcotubules, together with exceedingly small cell diameters (less than 5 µ), support the suggestion that the mechanism of excitation-contraction coupling differs in these muscle cells from that now thought to be characteristic of striated muscle such as skeletal muscle and mammalian cardiac muscle. These structural dissimilarities also imply that the mechanism of relaxation in frog ventricular muscle differs from that considered typical of other striated muscles. Additional ultrastructural features of frog ventricular heart muscle include spherical electron-opaque bodies on thin filaments, inconstantly present, forming a rank across the I band about 150 mµ from the Z line, and membrane-bounded dense granules resembling neurosecretory granules. The functional significance of these features is not yet clear.

scholarly journals Occurrence and immunolocalization of plectin in tissues.

1983 ◽  
Vol 97 (3) ◽  
pp. 887-901 ◽  
Author(s):  
G Wiche ◽  
R Krepler ◽  
U Artlieb ◽  
R Pytela ◽  
H Denk
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Urinary Bladder ◽  
Cardiac Muscle ◽  
Muscle Cells ◽  
Cell Types ◽  
Cell Junctions ◽  
Cell Periphery ◽  
Attachment Sites ◽  
Cytoplasmic Filaments

Various tissues from rat were examined for the occurrence and cellular localization of plectin, a 300,000-dalton polypeptide component present in intermediate filament-enriched cytoskeletons prepared from cultured cells by treatment with nonionic detergent and high salt solution. The extraction of liver, heart, skeletal muscle, tongue, and urinary bladder with 1% Triton/0.6 M KCl yielded insoluble cell residues that contained polypeptides of Mr 300,000 in variable amounts. These high Mr polypeptide species and a few bands of slightly lower Mr (most likely proteolytic breakdown products) were shown to react with antibodies to rat glioma C6 cell plectin using immunoautoradiography and/or immunoprecipitation. By indirect immunofluorescence microscopy using frozen sections (4 micron) of stomach, kidney, small intestine, liver, uterus, urinary bladder, and heart, antigens reacting with antibodies to plectin were found in fibroblast, endothelial, smooth, skeletal, and cardiac muscle, nerve, and epithelial cells of various types. Depending on the cell type, staining was observed either throughout the cytoplasm, or primarily at the periphery of cells, or in both locations. In hepatocytes, besides granular staining at the cell periphery, conspicuous staining of junctions sealing bile canaliculi was seen. In cardiac muscle strong staining was seen at intercalated disks and, as in skeletal muscle, at Z-lines. In cross sections through smooth muscle, most strikingly of urinary bladder, antibodies to plectin specifically decorated regularly spaced, spot-like structures at the cell periphery. By immunoelectron microscopy using the peroxidase technique, antiplectin-reactive material was found along cell junctions of hepatocytes and was particularly enriched at desmosomal plaques and structures associated with their cytoplasmic surfaces. A specific immunoreaction with desmosomes was also evident in sections through tongue. In cardiac muscle, besides Z-lines, intercalated disks were reactive along almost their entire surface, suggesting that plectin was associated with the fascia adherens, desmosomes, and probably gap junctions. In smooth muscle cells, regularly spaced lateral densities probably representing myofilament attachment sites were immunoreactive with plectin antibodies. The results show that plectin is of widespread occurrence with regard to tissues and cell types. Furthermore, immunolocalization by light and electron microscopy at junctional sites of various cell types and at attachment sites of cytoplasmic filaments in epithelial and muscle cells suggests that plectin possibly plays a universal role in the formation of cell junctions and the anchorage of cytoplasmic filaments.

Proteins of the contractile mechanism of mammalian smooth muscle and their possible location in the cell

1964 ◽  
Vol 160 (981) ◽  
pp. 517-524 ◽  
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Sedimentation Rate ◽  
Striated Muscle ◽  
Muscle Actin ◽  
Trypsin Treatment ◽  
Skeletal Muscle Myosin ◽  
Uterine Smooth Muscle ◽  
Muscle Myosin ◽  
Viscous Behaviour

Dorothy M. Needham speaking. Since the pioneer work of Csapo and his colleagues, beginning about fifteen years ago, it has been realized that from uterine smooth muscle can be extracted a protein closely resembling skeletal-muscle actomyosin in its viscous behaviour, sedimentation rate and electrophoretic mobility. (See, for example, Csapo 1948, 1949, 1950, 1959; Csapo, Erdos, Naeslund & Snellman 1950; Naeslund & Snellman 1951). Later work, in which the properties of purified preparations of myosin, actin and actomyosin have been studied, bears out these earlier conclusions. Thus, for example, we have shown (Needham & Williams 1963 b ) that skeletal-muscle myosin will react normally with uterus actin to give the highly viscous actomyosin; and similarly uterus myosin with skeletal-muscle actin. In both types of experiment the results indicated that the two proteins associated together in about the same proportions as when both are derived from skeletal muscle. Uterus actomyosin may be fragmented by carefully controlled trypsin treatment giving light and heavy meromyosins which, so far as they have been studied, show similar properties to the meromyosins from skeletal-muscle actomyosin (Needham & Williams 1959; Cohen, Lowey & Kucera 1961). Smooth muscle, however, does contain very strikingly less actomyosin than striated muscle, only 6 to 10 mg/g wet wt as compared with about 70 mg/g wet wt in skeletal muscle (Needham & Williams 1963 a ).

scholarly journals Multiple regulatory elements contribute differentially to muscle creatine kinase enhancer activity in skeletal and cardiac muscle.

1993 ◽  
Vol 13 (5) ◽  
pp. 2753-2764 ◽  
Author(s):  
S L Amacher ◽  
J N Buskin ◽  
S D Hauschka
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Muscle ◽  
Striated Muscle ◽  
Regulatory Elements ◽  
High Level Expression ◽  
Muscle Creatine Kinase ◽  
Skeletal Myocytes ◽  
E Box ◽  
Nonmuscle Cells ◽  
High Level

We have used transient transfections in MM14 skeletal muscle cells, newborn rat primary ventricular myocardiocytes, and nonmuscle cells to characterize regulatory elements of the mouse muscle creatine kinase (MCK) gene. Deletion analysis of MCK 5'-flanking sequence reveals a striated muscle-specific, positive regulatory region between -1256 and -1020. A 206-bp fragment from this region acts as a skeletal muscle enhancer and confers orientation-dependent activity in myocardiocytes. A 110-bp enhancer subfragment confers high-level expression in skeletal myocytes but is inactive in myocardiocytes, indicating that skeletal and cardiac muscle MCK regulatory sites are distinguishable. To further delineate muscle regulatory sequences, we tested six sites within the MCK enhancer for their functional importance. Mutations at five sites decrease expression in skeletal muscle, cardiac muscle, and nonmuscle cells. Mutations at two of these sites, Left E box and MEF2, cause similar decreases in all three cell types. Mutations at three sites have larger effects in muscle than nonmuscle cells; an A/T-rich site mutation has a pronounced effect in both striated muscle types, mutations at the MEF1 (Right E-box) site are relatively specific to expression in skeletal muscle, and mutations at the CArG site are relatively specific to expression in cardiac muscle. Changes at the AP2 site tend to increase expression in muscle cells but decrease it in nonmuscle cells. In contrast to reports involving cotransfection of 10T1/2 cells with plasmids expressing the myogenic determination factor MyoD, we show that the skeletal myocyte activity of multimerized MEF1 sites is 30-fold lower than that of the 206-bp enhancer. Thus, MyoD binding sites alone are not sufficient for high-level expression in skeletal myocytes containing endogenous levels of MyoD and other myogenic determination factors.

The Anatomy of the Sarcoplasmic Reticulum in Vertebrate Skeletal Muscle: Its Implications for Excitation Contraction Coupling

1982 ◽  
Vol 37 (7-8) ◽  
pp. 665-678 ◽  
Author(s):  
Joachim R. Sommer
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Action Potential ◽  
Cardiac Muscle ◽  
Calcium Release ◽  
Striated Muscle ◽  
Band Region ◽  
Terminal Cisterna ◽  
Transmitter Substance ◽  
Main Components

Abstract The sarcoplasmic reticulum in situ is an intricate tubular network that surrounds the contractile material in striated muscle cells. Its topographical relationship to other intracellular components, especially the myofibrils, is rather rigidly mainiained by a cytoskeleton which enmeshes Z line material and sarcoplasmic reticulum and, ultimately, is anchored at the plasmalemma. As a result, the two main components of the sarcoplasmic reticulum, the junctional SR and the free SR, retain their typical location in the A band region and in the I band region, respectively. The junc­tional SR, which is thought to be the site for calcium storage and release for contraction, is, thus, always well within one micron of the regulatory proteins associated with the actin filaments. The junctional SR, a synonym for terminal cisterna applying to both skeletal and cardiac muscle, is generally held to be involved in the translation of the action potential into calcium release, mainly because of the close topographic apposition between the junctional SR and the plasmalemma, especially in skeletal muscle. This attractive structure-function correlation is challenged by the observation that in bird cardiac muscle 80% of the junctional SR is spacially far removed from plas­malemma, the site of electrical activity. This anomalous topography is not in conflict with the notion that translation of the action potential into calcium release may be accomplished by a dif­fusible transmitter substance, e.g. calcium. Any hypothesis dealing with this problem must ac­ count for the anatomy of the bird heart.

scholarly journals A distinct cardiopharyngeal mesoderm genetic hierarchy establishes antero-posterior patterning of esophagus striated muscle

2019 ◽  
Author(s):  
Glenda Comai ◽  
Églantine Heude ◽  
Sebastien Mella ◽  
Sylvain Paisant ◽  
Francesca Pala ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Striated Muscle ◽  
Striated Muscles ◽  
Loss Of Function ◽  
Pharmacological Studies ◽  
Posterior Migration ◽  
Functional Relevance ◽  
Genetic Signatures ◽  
Genetic Hierarchy

SUMMARYIn most vertebrates, the upper digestive tract is composed of muscularised jaws linked to the esophagus that permit food uptake and swallowing. Masticatory and esophagus striated muscles (ESM) share a common cardiopharyngeal mesoderm (CPM) origin, however ESM are unusual among striated muscles as they are established in the absence of a primary skeletal muscle scaffold. Using mouse chimeras, we show that the transcription factors Tbx1 and Isl1 are required cell-autonomously for myogenic specification of ESM progenitors. Further, genetic loss-of-function and pharmacological studies point to Met/HGF signalling for antero-posterior migration of esophagus muscle progenitors, where HGF ligand is expressed in adjacent smooth muscle cells. These observations highlight the functional relevance of a smooth and striated muscle progenitor dialogue for ESM patterning. Our findings establish a Tbx1-Isl1-Met genetic hierarchy that uniquely regulate esophagus myogenesis and identify distinct genetic signatures that can be used as a framework to interpret pathologies arising within CPM derivatives.

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