Three-dimensional distributions of desmin and vimentin in cultured hamster cardiomyocytes using the immunogold deep-etching replica technique.
The distributions of desmin and vimentin intermediate filaments in cultured hamster heart cells were examined by immunofluorescent microscopy and an immunogold deep-etching replica technique in combination with electron microscopy. Fluorescent studies showed the overall staining patterns of the myocytes as well as the fibroblasts. Monoclonal antibodies (Da, D3) to desmin showed punctate staining for the myocytes, while polyclonal desmin (pD) stained in a filamentous pattern. Fibroblasts stained strongly with monoclonal anti-vimentin (Va), but did not stain with the desmin probes. Deep-etched immunogold studies confirmed at the ultrastructural level that monoclonal anti-desmin antibodies stain individual intermediate filaments in an intermittent pattern. Monoclonal (D3) antibody stained the intermediate filaments heavily and continuously at the cell peripheries, while it stained intermittently in the cell body, similar to the Da monoclonal. Monoclonal anti-vimentin stained only intermediate filaments in fibroblasts. Our studies show a heterogeneity of staining within the cultured heart cells when various anti-desmin and anti-vimentin antibodies are used.
Isobe
Y
Y
Department of Anatomy and Cell Biology, State University of New York, Health Science Center at Syracuse 13210.
Nakatsugawa
M
M
Hou
G R
GR
Lemanski
L F
LF
D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002478:Cells, Cultured; D006224:Cricetinae; D003893:Desmin / Q000032:analysis*; D005347:Fibroblasts / Q000166:cytology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D005613:Freeze Etching; D006046:Gold; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D009206:Myocardium / Q000166:cytology* / Q000648:ultrastructure; D014746:Vimentin / Q000032:analysis*
D000911:Antibodies, Monoclonal; D003893:Desmin; D014746:Vimentin; D006046:Gold