V-type H⁺ ATPase Activity is Required for Embryonic Dorsal-Ventral Symmetry Breaking

Abstract

The mechanism for embryonic dorsal-ventral (DV) symmetry breaking is idiosyncratic to the species, then converges on polarized expression of BMP signaling ligands. Here, we show that V-ATPase (VHA) activity is an early requirement for DV symmetry breaking in sea urchin embryos. In these basal deuterostomes, DV specification is mediated by ventral Nodal expression that leads to the establishment of a BMP signaling gradient. Nodal expression occurs downstream from p38 MAPK, which is transiently asymmetrically active. We show that VHA activity is required for DV symmetry breaking upstream from both p38 MAPK and Nodal. We rescue VHA-mediated ventralization by enforcing Nodal signaling asymmetry. We identify a VHA-dependent DV voltage gradient and also find that VHA activity is required for hypoxia inducible factor (HIF) activation. However, neither hyperpolarization nor HIF activation account for the dorsalizing effects of VHA, implicating a third unknown pathway that connects VHA activity to p38 MAPK symmetry breaking.