Partial oxygen pressure and mitochondrial permeability transition affect germ cell apoptosis in the human testis.

ABSTRACT

During regular spermatogenesis, a number of testicular germ cells degenerate by an apoptotic process that is under hormonal control. Oxidative and mitochondrial changes have been proposed to play a role in apoptosis of many cell types. Previously, whether human germ cell survival is controlled by oxygen or by effectors of the mitochondrial permeability transition has not been investigated. In the present study, apoptosis was induced in human testicular germ cells by incubating segments of seminiferous tubules without survival factors (i.e. serum or hormones; 21% oxygen). Apoptosis was significantly suppressed in an inversely dose-dependent fashion at partial oxygen pressures below 10%, as detected by Southern blot analysis of DNA fragmentation, DNA labeling in situ, and electron microscopy. Cyclosporin A and its nonimmunosuppressive derivative N-methyl-Val4-cyclosporin A prevented cell death, suggesting a key role for the mitochondrial permeability transition in apoptosis. Apoptotic cells were identified as mainly spermatocytes and spermatids, the mitochondria of which underwent morphological changes during the apoptotic process. The present results imply that to improve germ cell viability in in vitro fertilization techniques, the partial oxygen pressure should be lowered.