A cytochemical and immunocytochemical study of DNA distribution in spermatid nuclei of mouse, rabbit, and bull.
DNA distribution in mouse, rabbit and bull spermatids was analyzed by electron microscopy, after using a Feulgen-like HCl-osmium ammine procedure, and after immunocytochemistry with anti-DNA antibodies. In addition, nucleic acids were visualized with the intercalating dye ethidium bromide and phosphotungstic acid. The parts of DNA displaying a beta helix configuration (possibly A-T rich parts) were identified by epifluorescence microscopy after staining with Hoechst 33258. In all 3 species, young spermatid nuclei were seen to have large areas poor in DNA, as well as DNA-rich areas, which were mostly concentrated into a peripheral layer close to the acrosome and into one or several masses, displaying species-specific locations. These DNA-rich areas were stained with Hoechst 33258. Elongating spermatid nucleic contained homogeneously distributed DNA, and this was evident following both immunocytochemistry and nucleic acid histochemistry in all 3 species. However, the distribution appeared more heterogeneous after the Feulgen-like procedure, and was accompanied by a disappearance of Hoechst-fluorescence. In fully elongated spermatids, all nuclear areas stained with Hoechst 33258, while the 3 other techniques labeled either all or species-specific parts of the condensed chromatin. The reasons for these variable reactions are discussed in terms of technique specificities, DNA configuration and nucleoprotein moiety replacements.
Courtens
J L
JL
INRA, Physiologie de la Reproduction, Monnaie, France.
Biggiogera
M
M
Fakan
S
S
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000648:ultrastructure*; D004247:DNA / Q000032:analysis*; D006056:Golgi Apparatus / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D011817:Rabbits; D013087:Spermatids / Q000166:cytology / Q000648:ultrastructure*; D013194:Staining and Labeling
D004247:DNA