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J. Blanco-Rodríguez and C. Martínez-García

It is widely assumed that oestrogen administration in the male mimics hypophysectomy by suppressing gonadotrophin secretion. Nevertheless, oestradiol treatment can increase germ-cell apoptosis mainly at stages IV–X of the spermatogenic cycle, rather than at stage VII when apoptotic germ-cell death is mainly triggered by gonadotrophin withdrawal caused by hypophysectomy. Since the roles of testicular oestrogens in spermatogenic regulation, if any, are unknown, we re-evaluated the germ-cell types that undergo apoptosis after oestradiol treatment. Adult male rats were injected daily with 50 μg oestradiol, oestradiol plus testosterone propionate (25 mg every 3 days) or oestradiol plus human menopausal gonadotrophin (equivalent to 25 iu FSH plus 25 iu LH) for 15 days. Apoptosis was assessed by in situ 3′-end labelling of internucleosomal DNA fragments in plastic semithin sections; the germ-cell types involved were identified by high-resolution light microscopy. The quantitative analysis of our results shows that the apoptosis pattern elicited by oestradiol treatment of the seminiferous epithelium differs from that reported to be caused by gonadotrophin or testosterone withdrawal, suggesting a possible role for oestradiol in the modulation of germ-cell death in the adult testis of the rat.

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J Blanco-Rodriguez, C Martinez-Garcia and A Porras

In the seminiferous epithelium, both DNA synthesis and apoptosis occur at equivalent stages in various species, with apoptosis taking place mainly at the same stages as DNA replication in the second, third and fourth spermatogonial generations. As preservation of the cellular associations found at these stages may have some functional significance, it is important to determine whether there is a correlation between these cellular events. In this study, pairs of immunoperoxidase-stained adjacent testis sections from rats, mice, rabbits and cats in which either bromodeoxyuridine incorporated into the newly synthesized DNA strand (BrdU labelling) or DNA 3' end labelling of the apoptotic DNA fragments (TUNEL assay) were detected were compared. In addition, both events were analysed in double-labelled sections. These two methods revealed a clear correlation between the occurrence of DNA replication in the second to fourth generations of spermatogonia and most physiological apoptosis taking place in both spermatogonia and spermatocytes in the three different mammalian orders (Rodentia, Lagomorpha and Carnivora). This correlation may result from the synchronization of mitotic spermatogonial and meiotic spermatocyte cell cycle checkpoints operating at these stages.