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Abstract
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 to 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'-diisothiocyanostilbene-2,2'-disulfonic acid). We
remark that bicarbonate acts primarily through activation
of sAC in human spermatozoa, to stimulate tyrosine
phosphorylation of AKAP3 and sperm motility since 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 increased
recruitment of PKA to AKAP3.
Key words:
Gamete Biology •
Cyclic adenosine monophosphate •
Kinases •
Signal transduction •
Sperm motility and transport
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