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BOR - Papers in Press, published online ahead of print December 10, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.023085
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Submitted September 11, 2003
Returned for revision October 8, 2003
Accepted December 1, 2003

Male Reproductive Tract


Expression and Localization of Hypoxia-Inducible Factor-1 Subunits in the Adult Rat Epididymis

M. A. Palladino *, J. D. Powell , N. Korah , and L. Hermo

* To whom correspondence should be addressed. E-mail: mpalladi{at}monmouth.edu.

Abstract
The epididymal epithelium contributes to formation of a luminal fluid that is essential for the protection of spermatozoa from a variety of insults including changes in oxygen tension. A key regulator of the response to oxygen debt in many cells is hypoxia-inducible factor-1 (HIF-1). A transcription factor composed of {alpha} and {beta} subunits, HIF-1 activates genes that mediate oxygen homeostasis and cell survival pathways or trigger cell death responses. Previously we have shown that HIF-1{alpha} mRNA is expressed in the adult rat epididymis. Goals of this study were to determine if HIF-1{alpha} protein is activated by ischemia in the rat epididymis, to determine if epididymal HIF-1{alpha} mRNA expression is androgen-dependent, and to identify epididymal cell types expressing HIF-1{alpha} and {beta}. Immunoblot analysis revealed that HIF-1{alpha} protein is primarily present in corpus and cauda of the normoxic epididymis and unaffected by ischemia, while HIF-1{beta} was detected equally in all regions and also unaffected by ischemia. HIF-1{alpha} mRNA expression in all regions was not affected by 15 days bilateral orchiectomy. Principal cells stained positive for HIF-1{alpha} by immunocytochemistry, with the epithelium of initial segment and caput epididymidis staining less intensely than corpus and cauda. HIF-1{beta} immunoreactivity was equally present in principal cells in all regions. Clear, narrow and basal cells were unreactive for HIF-1{alpha} and {beta}. The presence of HIF-1 in normoxic epididymis and the regional distribution of HIF-1{beta} suggests fundamental differences in how proximal and distal regions of the epididymis maintain oxygen homeostasis to protect the epithelium and spermatozoa from hypoxia.

Key words: Male Reproductive Tract • Epididymis • Gene regulation • Sperm • Stress


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