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BOR - Papers in Press, published online ahead of print October 27, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.034520
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Submitted July 22, 2004
Returned for revision August 27, 2004
Accepted October 13, 2004

Testis


Functional Role of Caspases in Heat-Induced Testicular Germ Cell Apoptosis

Yanira Vera , Susana Rodriguez , Mark Castanares , Yanhe Lue , Vince Atienza , Christina Wang , Ronald S. Swerdloff , and Amiya P. Sinha Hikim *

* To whom correspondence should be addressed. E-mail: hikim{at}gcrc.rei.edu.

Abstract
In the present study, we determined whether a pan caspase inhibitor could prevent or attenuate heat-induced germ cell apoptosis. Groups of 5 adult (8 weeks old) C57BL/6 mice pre-treated with vehicle (DMSO) or Quinoline-Val-Asp (Ome)-CH2-O-Ph (Q-VD-OPH), a new generation broad-spectrum caspase inhibitor were exposed once to local testicular heating (43° for 15 min) and killed 6 h later. The inhibitor (40 mg/kg BW) or vehicle was administered intraperitoneally (ip) 1 h prior to local testicular heating. Germ cell apoptosis was detected by TUNEL assay and quantitated as number of apoptotic germ cells per 100 Sertoli cells at stages XI-XII. Compared with controls (16.8 ± 3.1), mild testicular hyperthermia within 6 h resulted in a marked activation (277.3 ± 21.6) of germ cell apoptosis, as previously reported by us. Q-VD-OPH at this dose markedly inhibited caspase 3 activation and significantly prevented (by 67.0%) heat-induced germ cell apoptosis. Q-VD-OPH-mediated rescue of germ cells was independent of cytosolic translocation of mitochondrial cytochrome c and DIABLO. Electron microscopy further revealed normal appearance of these rescued cells. Similar protection form heat-induced germ cell apoptosis was also noted after pretreatment with minocycline, a second generation tetracycline that effectively inhibits cytochrome c release and, in turn, caspase activation. Collectively, the present study emphasizes the role of caspases in heat-induced germ cell apoptosis.

Key words: Gamete Biology • Testis • Apoptosis • Spermatogenesis


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