Apoptosis precedes detachment of germ cells from the seminiferous epithelium after hormone suppression by short-term oestradiol treatment of rats

Spermatogenesis is a highly synchronized process in which FSH and testosterone are considered the major regulators. Nevertheless, the mechanism by which these hormones act on germ cells is unclear. Cell adhesion has been proved to play an essential role in the regulation of programmed cell death in epithelial cells and it is now known that FSH and testosterone withdrawal results in the triggering of apoptosis as well as germ cell detachment from the seminiferous epithelium. Therefore, it seemed important to investigate whether the triggering of apoptosis in germ cells by experimental hormone suppression occurred as a result of their previous detachment from the epithelium. To achieve this goal, adult male rats were injected with 50 μg oestradiol benzoate for 1, 2, 3, 4, 5 or 10 days to suppress gonadotrophin secretion and thus intratesticular levels of testosterone. Germ cell apoptosis was assessed in testes from these animals by in situ 3' end-labelling of DNA fragments and quantified in seminiferous tubule sections at stages VII–VIII. Serial sections throughout the epididymides from these animals were analysed to search for immature germ cells detached from the epithelium. These cells were scored and quantified in non-consecutive randomly selected epididymal sections. Our data indicate that the triggering of apoptosis in germ cells precedes germ cell detachment, suggesting that detachment of germ cells from the epithelium, occurring after hormone suppression, is not necessary for germ cell apoptosis.