Journal of Andrology, Vol. 24, No. 1, January/February 2003
Copyright © American Society of Andrology
Implication of Calmodulin-Dependent Phosphodiesterase Type 1 During Bovine Sperm Capacitation
VALÉRIE FOURNIER*,
PIERRE LECLERC
,
NATHALY CORMIER* AND
JANICE L. BAILEY*
From the * Centre de Recherche en Biologie de la
Reproduction, Département des sciences animales, and
Département d'obstétrique et
gynécologie, Université Laval, Québec, Québec
Canada.
| Correspondence to: Janice L. Bailey PhD, Centre de Recherche en Biologie de la
Reproduction, Département des sciences animales, Université
Laval, Québec, Québec G1K 7P4, Canada (e-mail:
janice.bailey{at}crbr.ulaval.ca). |
Phosphodiesterases (PDEs) are enzymes that degrade cyclic nucleotides. The
calcium-calmodulin dependent PDE type 1 (PDE 1) and the cyclic adenosine
monophosphate (cAMP)-specific PDE type 4 (PDE 4) have been implicated in sperm
function. We tested the hypothesis that specific PDEs regulate capacitation of
bovine sperm in a manner independent of those that mediate motility. Our
objectives were to determine the effects of inhibiting PDE 1 and PDE 4 on
capacitation and motility, and to compare these effects to those of heparin,
which is necessary for capacitation of bull sperm in vitro. Fresh sperm were
supplemented either with 15 µg/mL heparin (positive control) or the PDE
inhibitors vinpocetine (specific for PDE 1) and rolipram (specific for PDE 4),
and then incubated for 5 hours. At 0, 3, and 5 hours, samples were assayed for
capacitation and motility parameters according to the chlortetracycline (CTC)
fluorescent pattern B and computer-assisted sperm analysis, respectively. A
higher percentage of CTC pattern B sperm relative to heparin controls was
observed at 0 and 3 hours when sperm were incubated with vinpocetine. After 5
hours, the percentage of heparin- and vinpocetine-treated sperm showing
pattern B did not differ (P > .05). Rolipram did not affect CTC
patterns (P > .05; n = 4). Vinpocetine and heparin both reduced
the percentage of progressively motile sperm after 3 and 5 hours, but
vinpocetine reduced it more than heparin (P < .05; n = 4).
Rolipram transiently increased linearity versus sperm with heparin (P
< .05; n = 4). To further test the hypothesis that PDE 1 inhibition permits
capacitation, we conducted in vitro fertilization. Vinpocetine did not support
the ability of sperm to penetrate homologous oocytes (n = 5). Although cAMP
regulation by PDE 1 may occur early during capacitation, downstream events
appear to prevent full capacitation from occurring prematurely.
Key words: Vinpocetine, rolipram, cyclic AMP, in vitro fertilization, motility
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Copyright © 2003 by The American Society of Andrology.
