Ultrastructural localization of tubulin and actin in polyethylene glycol-embedded rat seminiferous epithelium by immunogold staining.
The polyethylene glycol (PEG) method for immunofluorescence localization of cytoskeletal antigens has been extended to the ultrastructural level using glutaraldehyde-fixed tissues and immunogold staining. Semithin sections of fixed tissue embedded in polyethylene glycol are divested of the PEG, exposed to purified antibodies (e.g., antiactin, antitubulin) and anti-IgG-colloidal gold. The sections may be processed by dehydration and critical-point drying, or reembedment in hydrophilic substances. Tubulin is demonstrated in the mitotic spindles of dividing spermatogonia, manchettes, axonemes and centrioles of developing spermatids, and in the Sertoli cell cytoplasm; actin localization is demonstrated in the myoid cells of the tunica propria, and smooth muscle cells of arterioles in the interstitial tissue. The results demonstrate the applicability and versatility of PEG embedding for immunocytochemistry.
D000199:Actins / Q000032:analysis*; D000818:Animals; D005455:Fluorescent Antibody Technique; D005976:Glutaral; D006046:Gold; D006651:Histocytochemistry / Q000379:methods*; D008297:Male; D008854:Microscopy, Electron; D011092:Polyethylene Glycols; D051381:Rats; D012670:Seminiferous Epithelium / Q000032:analysis* / Q000648:ultrastructure; D013194:Staining and Labeling; D013737:Testis / Q000032:analysis*; D014404:Tubulin / Q000032:analysis*
D000199:Actins; D014404:Tubulin; D011092:Polyethylene Glycols; D006046:Gold; D005976:Glutaral