3-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 cul tured hamster heart cells were examined by immunofluorescent microscop y 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 antibo dies (Da, D-3) to desmin showed punctate staining for the myocytes, wh ile polyclonal desmin (pD) stained in a filamentous pattern. Fibroblas ts stained strongly with monoclonal anti-vimentin (Va), but did not st ain with the desmin probes. Deep-etched immunogold studies confirmed a t the ultrastructural level that monoclonal anti-desmin antibodies sta in individual intermediate filaments in an intermittent pattern. Monoc lonal (D-3) antibody stained the intermediate filaments heavily and co ntinuously 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 vario us anti-desmin and anti-vimentin antibodies are used.