Intracellular chloride activity and pH during polar lobe formation and cytokinesis in eggs and embryos of Ilyanassa obsoleta.

Abstract

The objectives of this study were to compare membrane potential, intracellular chloride activity, and intracellular pH in unfertilized and fertilized eggs of Ilyanassa obsoleta throughout polar lobe formation and cytokinesis. Membrane potential, psi m, was measured with single- and double-barrel microelectrodes containing 3 M KCl. Chloride activity, (Cl)c, was measured with single-barrel, chloride-sensitive microelectrodes. Intracellular pH, pHc, was measured with double-barrel microelectrodes, utilizing an antimony-filled barrel to monitor pHc. Unfertilized eggs having a psi m of -13.7 mV displayed an average (Cl)c of 29.9 mM, whereas at electrochemical equilibrium, (Cl)ce, it would be 155.7 mM. This suggests that Cl is pumped out of the cell in eggs that are not fertilized. Fertilized eggs had a psi m of -78.2 mV and a (Cl)c of 18.6 mM. At electrochemical equilibrium, (Cl)ce would be 12.1 mM, thus suggesting that Cl is pumped into the cell in fertilized eggs. Double-barrel microelectrodes measured a psi m of -69.0 mV and a pHc of 7.3 in fertilized eggs. The psi m, (Cl)c, and pHc remained stable as fertilized eggs underwent the cellular shape changes involved in forming first, second, and third polar lobes and first cleavage.