Article: article from journal or magazin.
Ultrastructural localization of epidermal growth factor (EGF)-receptor transcripts in the cell nucleus using pre-embedding in situ hybridization in combination with ultra-small gold probes and silver enhancement.
A high-resolution in situ hybridization method is described for localizing epidermal growth factor (EGF)-receptor transcripts in nuclei of A431 epidermoid carcinoma cells. The method is based upon the use of ultra-small gold particles in combination with silver enhancement. The RNA of the EGF-receptor was detected mainly around the nucleoli. After removal of the DNA using nucleases and high salt extraction, the RNA of the EGF-receptor appears to be associated with the nuclear matrix. The RNA of the EGF-receptor was observed in close contact with the SC-35 splicing protein, but no exact colocalization was observed. These results demonstrate that high resolution pre-embedding in situ hybridization in combination with immunocytochemistry, both using ultra-small gold as a detection method, provides a powerful tool to unravel the organization of nuclear processes.
3T3 Cells, Animals, Carcinoma, Squamous Cell, Cell Line, Cell Nucleolus/metabolism, Cell Nucleolus/ultrastructure, Cell Nucleus/metabolism, Cell Nucleus/ultrastructure, Gold, Humans, In Situ Hybridization/methods, Mice, Microscopy, Electron/methods, Nuclear Matrix/metabolism, Nuclear Matrix/ultrastructure, RNA Probes, Receptor, Epidermal Growth Factor/analysis, Receptor, Epidermal Growth Factor/biosynthesis, Silver, Staining and Labeling, Tumor Cells, Cultured
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