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Division of Endocrinology, Departments of Medicine (A.P.S.H., Y.L., C.M.Y., Y.V., S.R., K.S., C.W., R.S.S.) and Pediatrics (P.H.Y.), Harbor-University of California-Los Angeles Medical Center and Research and Education Institute, Torrance, California 90509
Address all correspondence and requests for reprints to: Dr. Amiya P. Sinha Hikim, Division of Endocrinology, Harbor-University of California-Los Angeles Medical Center, Box 446, 1000 West Carson Street, Torrance, California 90509. E-mail: hikim{at}gcrc.rei.edu.
Short-term exposure (43 C for 15 min) of the rat testis to mild heat results within 6 h in stage- and cell-specific activation of germ cell apoptosis. Initiation of apoptosis was preceded by a redistribution of Bax from a cytoplasmic to paranuclear localization in heat-susceptible germ cells. Here we show that the relocation of Bax is accompanied by 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 poly(ADP) ribose polymerase. Furthermore, early in apoptosis, a significant amount of Bax also accumulates in endoplasmic reticulum, as assessed by Western blot analyses of fractionated testicular lysates. In additional studies using the FasL-defective gld mice, we have shown that heat-induced germ cell apoptosis is not blocked, thus providing evidence that the Fas signaling system may be dispensable for heat-induced germ cell apoptosis in the testis. Taken together, these results demonstrate that the mitochondria- and possibly also endoplasmic reticulum-dependent pathways are the key apoptotic pathways for heat-induced germ cell death in the testis.
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