Translator Disclaimer
6 April 2011 Guanine-Nucleotide Exchange Factors (RAPGEF3/RAPGEF4) Induce Sperm Membrane Depolarization and Acrosomal Exocytosis in Capacitated Stallion Sperm
L.A. McPartlin, P.E. Visconti, S.J. Bedford-Guaus
Author Affiliations +

Capacitation encompasses the molecular changes sperm undergo to fertilize an oocyte, some of which are postulated to occur via a cAMP-PRKACA (protein kinase A)-mediated pathway. Due to the recent discovery of cAMP-activated guanine nucleotide exchange factors RAPGEF3 and RAPGEF4, we sought to investigate the separate roles of PRKACA and RAPGEF3/RAPGEF4 in modulating capacitation and acrosomal exocytosis. Indirect immunofluorescence localized RAPGEF3 to the acrosome and subacrosomal ring and RAPGEF4 to the midpiece in equine sperm. Addition of the RAPGEF3/RAPGEF4-specific cAMP analogue 8-(p-chlorophenylthio)-2′-O-methyladenosine-3′,5′-cyclic monophosphate (8pCPT) to sperm incubated under both noncapacitating and capacitating conditions had no effect on protein tyrosine phosphorylation, thus supporting a PRKACA-mediated event. Conversely, activation of RAPGEF3/RAPGEF4 with 8pCPT induced acrosomal exocytosis in capacitated equine sperm at rates (34%) similar (P > 0.05) to those obtained in progesterone- and calcium ionophore-treated sperm. In the mouse, capacitation-dependent hyperpolarization of the sperm plasma membrane has been shown to recruit low voltage-activated T-type Ca2 channels, which later open in response to zona pellucida-induced membrane depolarization. We hypothesized that RAPGEF3 may be inducing acrosomal exocytosis via depolarization-dependent Ca2 influx, as RAPGEF3/RAPGEF4 have been demonstrated to play a role in the regulation of ion channels in somatic cells. We first compared the membrane potential (Em) of noncapacitated (−37.11 mV) and capacitated (−53.74 mV; P = 0.002) equine sperm. Interestingly, when sperm were incubated (6 h) under capacitating conditions in the presence of 8pCPT, Em remained depolarized (−32.06 mV). Altogether, these experiments support the hypothesis that RAPGEF3/RAPGEF4 activation regulates acrosomal exocytosis via its modulation of Em, a novel role for RAPGEF3/RAPGEF4 in the series of events required to achieve fertilization.

L.A. McPartlin, P.E. Visconti, and S.J. Bedford-Guaus "Guanine-Nucleotide Exchange Factors (RAPGEF3/RAPGEF4) Induce Sperm Membrane Depolarization and Acrosomal Exocytosis in Capacitated Stallion Sperm," Biology of Reproduction 85(1), 179-188, (6 April 2011).
Received: 26 May 2010; Accepted: 1 March 2011; Published: 6 April 2011

acrosome reaction
sperm capacitation
stallion sperm
Get copyright permission
Back to Top