Mechanism of intracellular pH increase during parthenogenetic activation of in vitro matured porcine oocytes

Parthenogenetic activation of porcine oocytes by using 7% ethanol, 50 or 100 mu M A23187 results in an increase in intracellular pH as does prolonged exposure to thimerosal. We attempt to specify which transporters or mechanisms are involved in the observed increase in intracellular pH during oocyte activation. Experiments were performed in the absence of sodium; the presence of 2.5 mM amiloride, a potent inhibitor of the Na+/ HC antiport; in the absence of bicarbonate; and in the presence of 4,4'-diisothiocyanafodihydrostilbene-2,2 acid, disodium salt (H2DIDS) for all three activation methods. These treatments had no effect on the increase in intracellular pH induced by the calcium ionophore or thimerosal, but all reduced the increase in pH (P < 0.001) in the 7% ethanol group. This suggests that the Na+/H+ antiport and the HCO3-/Cl- exchangers are not playing a role during treatment with calcium ionophore or thimerosal, and the pH increase observed during treatment with 7% ethanol may be dependent upon a sodium or bicarbonate flux (or both) into the oocyte. Bafilomycin Al (500 nm), an inhibitor of vacuolar-type H+ ATPases, had no effect on 7% ethanol or thimerosal treatments, but significantly reduced the increase in intracellular pH observed during calcium ionophore treatment. This may be the result of an initial local increase in intracellular free calcium levels.