Hum. Reprod. Advance Access published online on August 25, 2005
Human Reproduction, doi:10.1093/humrep/dei267
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1 National Hellenic Research Foundation, Institute of Biological Research and Biotechnology, Laboratory of Molecular Endocrinology, 48 Vas Constantinou Ave, 11635 Athens, Greece
* To whom correspondence should be addressed. BACKGROUND: The central role of estrogens and androgens in the male reproductive system has focused attention on the presence and distribution of their cognate receptors [estrogen receptor (ER)
Received March 2, 2005
Revised July 8, 2005
Accepted July 12, 2005
Article
Estrogen receptors
and 
(ER and ER) and androgen receptor (AR) in human sperm: localization of ER and AR in mitochondria of the midpiece
2 Foundation for Biomedical Research of the Academy of Athens, Center for Basic Research, Laboratory of Biochemistry, 4 Soranou Efesiou, 11527 Athens, Greece
3 IVF and Genetics, 296 Kifisias Ave and 40 Navarinou St, 15232 Athens, Greece
C.E. Sekeris, E-mail: csekeris{at}eie.gr
Abstract 
, ER
and androgen receptor (AR)] in male reproductive tissues and cells. Since the presence of steroid hormone receptors in mitochondria of mammalian cells has been well documented, we investigated the possibility of mitochondrial localization of sex steroid hormone receptors in sperm. METHODS AND RESULTS: Applying immunofluorescence labelling and confocal laser scanning microscopy we show that the estrogen receptor and the AR of human sperm are specifically enriched in the midpiece, at the site of the mitochondria, which were visualized by labelling with the vital dye CMX. Nuclear and mitochondrial localization of AR was also detected in LnCap human prostate cancer cells. Differentially, most of the ER immunostaining is in the form of a compact zone at a region corresponding to the equatorial segment of the upper post-acrosomal region of the sperm head. Immunoblotting experiments using sperm extracts revealed the presence of a 66 and a 45 kDa protein reacting with the ER antibody, one 64 kDa protein reacting with the ER antibody and a 110 and a 90 kDa protein reacting with the antibody against AR. CONCLUSIONS: Our findings suggest that the differential localization of AR and ER isoforms in human sperm reveals distinct roles of these receptors in the physiology of sperm cells and, perhaps, also in the process of fertilization./estrogen receptor /mitochondrion/sperm.
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