Ultrastructural localization of nuclear antigens during interphase in mouse 3T3 fibroblasts.
Thin sections of mouse 3T3 fibroblast nuclei labelled by immunoperoxidase with anti-nuclear antibodies I1, PI1, PI2, anti-peripherin, -lamin, and -centromere have been examined in the electron microscope. Staining was compared with the corresponding immunofluorescence labelling patterns, and was correlated with nuclear ultrastructure in conventionally fixed and uranyl-lead stained samples and in unlabelled immunoperoxidase controls. Peripherin was detected at the nuclear rim in a band broader and more irregular than the lamins/lamina. The peripheral components of PI1 and PI2 appear to be localized at nuclear pores and the nuclear envelope, respectively. The internal component of PI1 staining consisted of irregular patches and strands in the nucleoplasm, closely resembling snRNP staining as reported by others. Internal P12 labelling consisted of spots distributed apparently at random in interchromatinic regions. The spots resembled labelling by antibody I1, but were fewer and more irregular in size. Neither the PI2 nor the I1 spots were centromere associated, nor could they be correlated with specific interchromatinic structures in conventional preparations and in unlabelled controls. The results support the hypothesis that the nucleus is segregated into function-specific domains, distinguished by morphology and (or) composition from surrounding regions of the nucleus.
Chaly
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Ottawa-Carleton Resource Centre for Biological Microscopy, Department of Biology, Carleton University, Canada.
St Aubin
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Brown
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D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis*; D002467:Cell Nucleus / Q000276:immunology*; D002478:Cells, Cultured; D002503:Centromere / Q000276:immunology; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D007381:Intermediate Filament Proteins / Q000276:immunology; D007399:Interphase; D034882:Lamins; D008562:Membrane Glycoproteins; D051379:Mice; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins / Q000032:analysis / Q000276:immunology; D064531:Peripherins
D000911:Antibodies, Monoclonal; D000941:Antigens; D007381:Intermediate Filament Proteins; D034882:Lamins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D064531:Peripherins