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1 November 2000 Molecular Pathway of Germ Cell Apoptosis Following Ischemia/Reperfusion of the Rat Testis
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The present study investigates the molecular apoptotic pathway in germ cells following acute ischemia of the rat testis. Rats were subjected to ischemia-inducing torsion and testes were harvested after reperfusion. Apoptotic cells were identified with an antibody to single-stranded DNA. Seminiferous tubule RNA was examined by RNase protection assay or by reverse transcriptase-polymerase chain reaction (RT-PCR) for the presence and regulation of apoptotic molecules. Proteins from seminiferous tubules were used for Western blot analysis of cytochrome c. Germ cell apoptosis was maximal at 24 h after repair of torsion. Germ cells in stages II–III of the seminiferous epithelium cycle were the predominant early responders. The RNase protection assays revealed that Bcl-XL was the prominent mRNA species. Caspases 1, 2, 3, and Bax mRNA were consistently upregulated; however, the time of upregulation after torsion was variable. The Bcl-XL and Bcl-XS mRNAs were less consistently upregulated and there was no evidence for upregulation of Fas or Bcl-2. Fas ligand (FasL) was not detected by RNase protection assay, but RT-PCR revealed a significant increase in FasL expression 4 h after the repair of torsion. Western blot analysis for cytochrome c release demonstrated a significant increase 4 h after the repair of torsion. Results suggest that germ cell apoptosis following ischemia/reperfusion of the rat testis is initiated through the mitochondria-associated molecule Bax as well as Fas-FasL interactions.

Jeffrey J. Lysiak, Stephen D. Turner, and Terry T. Turner "Molecular Pathway of Germ Cell Apoptosis Following Ischemia/Reperfusion of the Rat Testis," Biology of Reproduction 63(5), 1465-1472, (1 November 2000).
Received: 21 February 2000; Accepted: 1 June 2000; Published: 1 November 2000

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