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Mol Reprod Dev ; 59(3): 314-21, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11424217

ABSTRACT

The nonchromatin structure or nuclear matrix in developing spermatogenic cells of the rat was studied using a biochemical fractionation in concert with resinless section electron microscopy. Observations demonstrated that the nuclear matrix of spermatogenic cells consisted of a three-dimensional network of filaments of variable thicknesses. In spermatogonia and spermatocytes the nuclear matrix consisted of relatively thin filaments, while that of round spermatids consisted of a thicker interconnecting network of filament. In elongating spermatids, the interior of the nuclear matrix consisted of a network of dense filaments bounded by a peripheral lamina. The protein composition of the nuclear matrix in spermatogenic cells was examined by high-resolution two-dimensional gel electrophoresis and correlated with morphological changes characteristic of each stage. The results showed that the proteins of nuclear matrix changed in a cell stage-specific manner. These stage-specific changes corresponded to the major transitions of chromatin structure and function during spermatogenesis. Furthermore, immunocytochemical and immunoblotting analysis of DNA topoisomerase II (topo II) revealed that this enzyme exhibited stage-specific variations and was associated with the nuclear matrix. These results suggest that the nuclear matrix in spermatogenic cells may be involved in mediating DNA modifications and maintaining nuclear organization during spermatogenesis. Mol. Reprod. Dev. 59:314-321, 2001.


Subject(s)
Nuclear Matrix/ultrastructure , Spermatogenesis , Spermatozoa/ultrastructure , Testis/cytology , Animals , Antigens, Nuclear , Cell Fractionation , DNA Topoisomerases, Type II/metabolism , Electrophoresis, Gel, Two-Dimensional , Histocytological Preparation Techniques/methods , Immunoblotting , Male , Microscopy, Immunoelectron , Nuclear Matrix/chemistry , Nuclear Proteins/analysis , Rats , Spermatids/chemistry , Spermatids/enzymology , Spermatids/ultrastructure , Spermatocytes/chemistry , Spermatocytes/enzymology , Spermatocytes/ultrastructure , Spermatogonia/chemistry , Spermatogonia/enzymology , Spermatogonia/ultrastructure , Spermatozoa/chemistry , Spermatozoa/enzymology , Testis/enzymology
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