Morphological integrity of single adult cardiac myocytes isolated by collagenase treatment: immunolocalization of tubulin, microtubule-associated proteins 1 and 2, plectin, vimentin, and vinculin.
Single cardiac myocytes were isolated from hearts of 9 to 12-week-old rats by means of collagenase (100 U/ml). After assessment of their functional integrity they were processed for immunofluorescence microscopy of the cytoskeletal proteins tubulin, microtubule-associated proteins 1 and 2 (MAP-1 and MAP-2), plectin, vimentin, and vinculin. Antibodies to tubulin decorated a delicate filamentous network that apparently was unrelated to any sarcomeric organization. The distribution of MAP-1 and MAP-2 was strikingly different from that of tubulin, as both antigens were confined to Z-line structures. These structures were also prominently stained by affinity-purified antibodies to plectin and a monoclonal antibody to vimentin. Co-distribution of plectin and vimentin was also observed at the former intercalated disk region of the heart cell. Anti-vinculin antibodies decorated an intricate meshwork consisting of delicate filaments with predominantly irregular orientation and occasional assembly into whorls. These immunolocalization data indicate that the cell shape and cytoskeletal architecture characteristic of cardiac myocytes in tissues is maintained in single isolated cells. Furthermore, intermediate filaments rather than microtubules seem to be instrumental in the preservation of cell morphology.
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D000818:Animals; D003599:Cytoskeleton / Q000648:ultrastructure; D004533:Egtazic Acid / Q000494:pharmacology; D005260:Female; D005455:Fluorescent Antibody Technique; D007381:Intermediate Filament Proteins; D008297:Male; D003012:Microbial Collagenase; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000648:ultrastructure; D009124:Muscle Proteins / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology* / Q000648:ultrastructure; D051190:Plectin; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D012518:Sarcomeres / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism*; D014746:Vimentin / Q000378:metabolism*; D016596:Vinculin
D007381:Intermediate Filament Proteins; D008869:Microtubule-Associated Proteins; D009124:Muscle Proteins; C495334:Plec protein, rat; D051190:Plectin; D011506:Proteins; D014404:Tubulin; D014746:Vimentin; D016596:Vinculin; D004533:Egtazic Acid; D003012:Microbial Collagenase