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Three-dimensional distributions of desmin and vimentin in cultured hamster cardiomyocytes using the immunogold
Y Isobe1, M Nakatsugawa, G R Hou
1Department of Anatomy and Cell Biology, State University of New York, Health Science Center at Syracuse 13210.
Insights
Investigating desmin and vimentin intermediate filaments in hamster heart cells revealed varied staining patterns. Different antibodies showed distinct distributions, highlighting cellular heterogeneity in desmin and vimentin expression.
Area of Science:
- Cell Biology
- Biochemistry
- Immunology
Background:
- Intermediate filaments, including desmin and vimentin, are crucial cytoskeletal components in various cell types.
- Understanding their distribution is vital for comprehending cellular structure and function, particularly in cardiomyocytes and fibroblasts.
Purpose of the Study:
- To investigate the precise distribution of desmin and vimentin intermediate filaments in cultured hamster heart cells.
- To compare the staining patterns obtained using different antibodies against desmin and vimentin.
Main Methods:
- Immunofluorescent microscopy was employed to visualize intermediate filament distribution.
- Immunogold deep-etching replica technique combined with electron microscopy provided ultrastructural details.
- Monoclonal and polyclonal antibodies specific to desmin and vimentin were utilized.
Main Results:
- Monoclonal anti-desmin antibodies (Da, D3) exhibited punctate staining in myocytes, while polyclonal anti-desmin (pD) showed a filamentous pattern.
- Monoclonal anti-vimentin antibodies (Va) strongly stained fibroblasts but not myocytes.
- Ultrastructural analysis confirmed intermittent staining of intermediate filaments by monoclonal anti-desmin antibodies, with variations between cell periphery and body.
Conclusions:
- Heterogeneity in desmin and vimentin intermediate filament distribution exists within cultured heart cells.
- The choice of antibody significantly influences the observed staining patterns, underscoring the importance of antibody specificity in intermediate filament research.
- These findings contribute to a deeper understanding of cytoskeletal organization in cardiac cells and fibroblasts.
Abstract:
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.