regulator of g protein signaling 3 modulates wnt5b calcium dynamics and somite patterning监管机构的3 g蛋白信号调节wnt5b钙动力学和体节模式.pdfVIP

Regulator of G Protein Signaling 3 Modulates Wnt5b Calcium Dynamics and Somite Patterning 1 2 1 Christina M. Freisinger , Rory A. Fisher , Diane C. Slusarski * 1 Department of Biology, University of Iowa, Iowa City, Iowa, United States of America, 2 Department of Pharmacology, University of Iowa College of Medicine, Iowa City, Iowa, United States of America Abstract Vertebrate development requires communication among cells of the embryo in order to define the body axis, and the Wnt- signaling network plays a key role in axis formation as well as in a vast array of other cellular processes. One arm of the Wnt- signaling network, the non-canonical Wnt pathway, mediates intracellular calcium release via activation of heterotrimeric G proteins. Regulator of G protein Signaling (RGS) proteins can accelerate inactivation of G proteins by acting as G protein GTPase-activating proteins (GAPs), however, the possible role of RGS proteins in non-canonical Wnt signaling and development is not known. Here, we identify rgs3 as having an overlapping expression pattern with wnt5b in zebrafish and reveal that individual knockdown of either rgs3 or wnt5b gene function produces similar somite patterning defects. Additionally, we describe endogenous calcium release dynamics in developing zebrafish somites and determine that both rgs3 and wnt5b function are required for appropriate frequency and amplitude of calcium release activity. Using rescue of gene knockdown and in vivo calcium imaging assays, we demonstrate that the activity of Rgs3 requires its ability to interact with Ga subunits and function as a G protein GAP. Thus, Rgs3 function is necessary for appropriate frequency and amplitude of calcium release during