Variations in dystrophin complex in red and white caudal muscles from Torpedo marmorata.
We present an up-to-date study on the nature, at the protein level, of various members of the dystrophin complex at the muscle cell membrane by comparing red and white caudal muscles from Torpedo marmorata. Our investigations involved immunodetection approaches and Western blotting analysis. We determined the presence or absence of different molecules belonging to the dystrophin family complex by analyzing their localization and molecular weight. Specific antibodies directed against dystrophin, i.e., DRP2 alpha-dystrobrevin, beta-dystroglycan, alpha-syntrophin, alpha-, beta-, gamma-, and delta-sarcoglycan, and sarcospan, were used. The immunofluorescence study (confocal microscopy) showed differences in positive immunoreactions at the sarcolemmal membrane in these slow-type and fast-type skeletal muscle fibers. Protein extracts from T. marmorata red and white muscles were analyzed by Western blotting and confirmed the presence of dystrophin and associated proteins at the expected molecular weights. Differences were confirmed by comparative immunoprecipitation analysis of enriched membrane preparations with anti-beta-dystroglycan polyclonal antibody. These experiments revealed clear complex or non-complex formation between members of the dystrophin system, depending on the muscle type analyzed. Differences in the potential function of these various dystrophin complexes in fast or slow muscle fibers are discussed in relation to previous data obtained in corresponding mammalian tissues. (J Histochem Cytochem 49:857-865, 2001)
Royuela
M
M
Department of Cell Biology and Genetics, University of Alcalá, Alcalá de Henares, Madrid, Spain. ricardo.paniagua@uah.es
Hugon
G
G
Rivier
F
F
Fehrentz
J A
JA
Martinez
J
J
Paniagua
R
R
Mornet
D
D
D000818:Animals; D015153:Blotting, Western; D002135:Calcium-Binding Proteins; D003598:Cytoskeletal Proteins / Q000378:metabolism; D049030:Dystroglycans; D016189:Dystrophin / Q000378:metabolism*; D049029:Dystrophin-Associated Proteins; D008562:Membrane Glycoproteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018656:Muscle Fibers, Fast-Twitch / Q000378:metabolism*; D009124:Muscle Proteins / Q000378:metabolism; D011233:Precipitin Tests; D049031:Sarcoglycans; D014101:Torpedo
D002135:Calcium-Binding Proteins; D003598:Cytoskeletal Proteins; D016189:Dystrophin; D049029:Dystrophin-Associated Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009124:Muscle Proteins; D049031:Sarcoglycans; C098885:dystrobrevin; C091825:syntrophin alpha1; D049030:Dystroglycans