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BOR - Papers in Press, published online ahead of print September 1, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.032490
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BIOLOGY OF REPRODUCTION 72, 22–32 (2005)
DOI: 10.1095/biolreprod.104.032490
© 2005 by the Society for the Study of Reproduction, Inc.

Tyrosine Phosphorylation of the A Kinase Anchoring Protein 3 (AKAP3) and Soluble Adenylate Cyclase Are Involved in the Increase of Human Sperm Motility by Bicarbonate1

M. Luconi2,3, I. Porazzi2, P. Ferruzzi4, S. Marchiani3, G. Forti3, and E. Baldi3

Departments of Clinical Physiopathology, Andrology,3 Endocrinology4 Units, Center for Research, Transfer and High Education MCIDNENT, University of Florence, I-50139 Florence, Italy

Mammalian testicular spermatozoa are immotile, thus, to reach the oocyte, they need to acquire swimming ability under the control of different factors acting during the sperm transit through the epididymis and the female genital tract. Although bicarbonate is known to physiologically increase motility by stimulating soluble adenylate cyclase (sAC) activity of mammalian spermatozoa, no extensive studies in human sperm have been performed yet to elucidate the additional molecular mechanisms involved. In this light, we investigated the effect of in vitro addition of bicarbonate to human spermatozoa on the main intracellular signaling pathways involved in regulation of motility, namely, intracellular cAMP production and protein tyrosine phosphorylation. Bicarbonate effects were compared with those of the phosphatidyl-inositol-3 kinase inhibitor, LY294002, previously demonstrated to be a pharmacological stimulus for sperm motility. Bicarbonate addition to spermatozoa results in a significant increase in sperm motility as well as in several hyperactivation parameters. This stimulatory effect of bicarbonate and LY294002 is mediated by an increase in cAMP production and tyrosine phosphorylation of the A kinase anchoring protein, AKAP3. The specificity of bicarbonate effects was confirmed by inhibition with 4,4'-di-isothiocyanostilbene-2,2'-disulfonic acid. We remark that, in human spermatozoa, bicarbonate acts primarily through activation of sAC to stimulate tyrosine phosphorylation of AKAP3 and sperm motility because both effects are blunted by the sAC inhibitor 2OH-estradiol. In conclusion, our data provide the first evidence that bicarbonate stimulates human sperm motility and hyperactivation through activation of sAC and tyrosine phosphorylation of AKAP3, finally leading to an increased recruitment of PKA to AKAP3.

1 Supported by grants from Italian Ministry of Education and Research (MIUR-COPIN) and the University of Florence.

2 Correspondence: Michaela Luconi, Unità di Andrologia, Dipartimento di Fisiopatologia Clinica, Università di Firenze, Viale Pieraccini 6, I-50139 Florence, Italy. FAX: 39 055 427 1371; m.luconi{at}dfc.unifi.it




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