Abstract
We have demonstrated that testosterone-bovine serum albumin conjugate labeled with 2-nm colloidal gold (testosterone-BSA-gold) injected into the vascular system of rat becomes visible as silver deposits on the sections of tissues embedded in epoxy resin after silver enhancement and enters the androgen-target cell nuclei, e.g. round spermatids, but not the non-target cell nuclei. The diameter of the silver deposits depends on the duration of silver enhancement. In this study, to make clear the transfer route of testosterone-BSA-gold into the round spermatid nucleus, the testis of rat injected testosterone-BSA-gold was observed under electron microscope after silver enhancement for short periods of time. The small silver deposits were present on the cell membrane, vesicles, Golgi region, acrosome, subacrosomal space, both the post-acrosomal and the subacrosomal nuclear envelope, and the nucleoplasm in the cap-phase spermatids. The silver deposits were also found in the perinuclear cisterna of post-acrosomal nuclear envelope, but not in the nuclear pore. When the spermatids were observed at high-power magnification without the silver enhancement, the outer nuclear membrane showed many irregular invaginations toward the inner nuclear membrane in the post-acrosomal nuclear envelope. Furthermore, a double-membrane-like vesicle seemed to be present in the nuclear envelope. In the vesicle, the gold particles were present along the inner membrane. These results suggest that testosterone-BSA-gold can enter the nucleoplasm through some route provided by the nuclear double membranes in both the post-acrosomal and the subacrosomal nuclear enve-lope.
