Molecular Human Reproduction, Vol. 9, No. 3, 117-124,
March 2003
© 2003 European Society of Human Reproduction and Embryology
Article |
Expression of aromatase in human ejaculated spermatozoa: a putative marker of motility
Submitted on January 23, 2002; resubmitted on September 10, 2002. accepted on November 26, 2002
1 UPRES EA 2608USC INRA, University-IRBA and 2 Department of Genetic and Reproduction, CHRU Clemenceau, Caen 14032, France
3 To whom correspondence should be addressed at: UPRES EA 2608USC INRA, Université-IRBA, Esplanade de la Paix, 14032 Caen Cedex, France. e-mail: carreau{at}ibba.unicaen.fr
Cytochrome P450 aromatase (P450arom) is a key enzyme responsible for the irreversible transformation of androgens into estrogens. In the present study, we have analysed the ability of human ejaculated spermatozoa to produce estrogens and for that purpose we have looked for the expression of specific aromatase transcript and protein. We have confirmed the presence of P450arom transcript in all normospermic purified samples by nested PCR. The sequence of PCR products from purified spermatozoa shares 98% identity with published human P450arom sequence. Using a semi-quantitative approach, we have observed in immotile sperm a significant decrease (28%) of the aromatase/glyceraldehyde-3-phosphate dehydrogenase ratio compared with the motile sperm fraction. On Western blot with a monoclonal antibody directed against aromatase, we have detected two bands (53 and 49 kDa) in microsome preparations from purified spermatozoa. In total protein extracts of purified spermatozoa (with and without cytoplasmic droplets), we have only found the aromatase as a 49 kDa band with a stronger intensity when cytoplasmic droplets are present. Moreover, the band seems to be weaker in immotile spermatozoa (with and without cytoplasmic droplets). Our data demonstrate the expression of aromatase both in terms of mRNA and protein in each sample of human purified spermatozoa and in addition, our results suggest that aromatase could be concerned with the acquisition of sperm motility.
Key words: aromatase/human spermatozoa/motility/mRNA/protein
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