Int. J. Dev. Biol. 39: 485 - 491 (1995)
4-aminopyridine acts as a weak base and a Ca2+ mobilizing agent in triggering oocyte meiosis reinitiation and activation in the Japanese clam Ruditapes philippinarum
Published: 1 June 1995
Abstract
Ovarian oocytes of the prosobranch mollusc Patella vulgata and the pelecypod Ruditapes philippinarum are arrested during prophase of the first maturation division. Release from this blockade, which is revealed by germinal vesicle breakdown, drives these oocytes to a second arrest in metaphase I, at which time the oocytes become fertilizable. The respective roles of Ca2+ and H+ ion movements during this early step in meiosis reinitiation has not been fully established yet. In this work we reveal the presence of acidic vesicles and report that bafilomycin A1 and N,N'-dicyclohexylcarbodiimide, two inhibitors of the vacuolar-type H(+)-ATPase, applied to Ruditapes oocytes, produce a significant inhibition of their response to the natural neurohormone serotonin. Since sodium deprivation did not affect this response, this suggests that a v-type ATPase pump, possibly located in the membrane of these acidic vesicles, may play a subtle role in the cascade of events that releases oocytes from their prophase block. We then describe how 4-aminopyridine, a drug reputed to be a K+ channel antagonist, triggers both meiosis reinitiation and activation of Patella and Ruditapes oocytes. This agent acts as a weak base, its effect depending on external pH. Moreover, using the fluorescent probes BCECF and Fluo-3/AM, we observe that this drug both alkalinizes the endoplasm and promotes an intracellular Ca2+ surge. This dual effect may explain why Ruditapes oocytes no longer stop in metaphase under these conditions and behave like other bivalve species which are directly fertilizable at the germinal vesicle stage.