Deciphering the pathways of germ cell apoptosis in the testis.
A growing body of evidence demonstrates that germ cell death both spontaneous (during normal spermatogenesis) and that induced by suppression of hormonal support or increased scrotal temperature occurs via apoptosis. The mechanisms by which these proapoptotic stimuli activate germ cell apoptosis are not well understood. In order to provide some insight, here we report the key molecular components of the effector pathways leading to caspase activation and increased germ cells apoptosis triggered by mildly increased scrotal temperature. Short-term exposure (43 degrees C for 15 min) of the testis to mild heat results, within 6h, in stage- and cell-specific activation of germ cell apoptosis in rats. Initiation of apoptosis was preceded by a redistribution of Bax from a cytoplasmic to paranuclear localization in heat-susceptible germ cells. Such relocation of Bax is further accompanied by sequestration of mitochondria and endoplasmic reticulum (ER) into paranuclear areas, cytosolic translocation of cytochrome c and is associated with activation of the initiator caspase 9 and the executioner caspases 3, 6, and 7, and cleavage of PARP. Furthermore, Bax is co-localized with ER in the susceptible germ cells as assessed by combined two-photon and confocal microscopy and Western blot analyses of fractionated testicular lysates. In additional studies, using gld and lpr(cg) mice, which harbor loss-of-function mutations in Fas-ligand (FasL) and Fas, respectively, we demonstrated that heat-induced germ cell apoptosis is not blocked, thus providing further evidence that the Fas signaling system is dispensable for heat-induced germ cell apoptosis in the testis. Taken together, these results demonstrate that the mitochondria- and possibly also ER-dependent pathways are the key apoptotic pathways for heat induced germ cell death in the testis.
Sinha Hikim
Amiya P
AP
Division of Endocrinology, Department of Medicine, Harbor-UCLA Medical Center and Research and Education Institute, Box 446, 1000W Carson Street, Torrance, CA 90509, USA. hilim@gcrc.rei.edu
Lue
Yanhe
Y
Diaz-Romero
Maruja
M
Yen
Pauline H
PH
Wang
Christina
C
Swerdloff
Ronald S
RS
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D006801:Humans; D008297:Male; D013090:Spermatocytes / Q000648:ultrastructure; D013094:Spermatozoa / Q000166:cytology* / Q000502:physiology; D013737:Testis / Q000166:cytology* / Q000502:physiology; D019014:fas Receptor / Q000502:physiology
D019014:fas Receptor