Ca2+ released via IP3 receptors is required for furrow deepening during cytokinesis in zebrafish embryos

Abstract

We have previously visualized three Ca2+ transients, generated by release from intracellular stores, which are associated with cytokinesis during the early cell division cycles of zebrafish embryos: the furrow positioning, propagation and deepening transients. Here we demonstrate the requirement of the latter for furrow deepening, and identify the Ca2+ release channels responsible for generating the deepening transient. The introduction of the Ca2+ buffer 5,5'-dibromo-BAPTA, at an appropriate time to challenge only the deepening transient, resulted in the dissipation of this transient and an inhibition of furrow deepening. Introduction of antagonists of the inositol 1,4,5-trisphosphate (IP3) receptor (heparin and 2-aminoethoxydiphenylborate; 2-APB) at the appropriate time, blocked the furrow deepening transient and resulted in an inhibition of furrow deepening. In contrast, antagonists of the ryanodine receptor and the NAADP-sensitive channel had no effect on either the furrow deepening transient or on furrow deepening. In addition, microinjection of IP3 led to the release of calcium from IP3-sensitive stores, whereas the introduction of caffeine or cADPR failed to induce any increase in intracellular Ca2+. Our new data thus support the idea that Ca2+ released via IP3 receptors is essential for generating the furrow deepening transient and demonstrate a requirement for a localized cytosolic Ca2+ riseforthe furrow deepening process. We also present data to show that the endoplasmic reticulum and IP3 receptors are localized on either side of the cleavage furrow, thus providing the intracellular Ca2+ store and release mechanism for generating the deepening transient.