Cell Adhesion and Spreading Affect Adipogenesis from Embryonic Stem Cells: The Role of Calreticulin

Abstract

Calreticulin is an endoplasmic reticulum-resident multifunctional protein, which has been shown to influence numerous cellular processes, including cell adhesion. In this study, we characterized the adhesive properties of embryonic stem cells (ESCs) lacking calreticulin and showed that adipogenesis from ESCs is directly and reciprocally controlled by the adhesive status of a cell, which in turn is modulated by calreticulin. Calreticulin-deficient ESCs are not only highly adipogenic but also show elevated calmodulin/CaMKII signaling and poor adhesiveness compared with the wild-type ESCs. Calreticulin deficiency leads to a disorganized cytoskeleton and low levels of focal adhesion-related proteins, such as vinculin, paxillin, and phosphorylated focal adhesion kinase, which cause limited focal adhesion formation and limited fibronectin deposition. Moreover, differentiation on nonadhesive substrata, which hinder cell spreading, promoted adipogenesis in the wild-type ESCs that normally have low adipogenic potential, causing a decrease in focal adhesion protein expression and an increase in calmodulin/CaMKII signaling. In contrast, inhibition of CaMKII effectively increased focal adhesion protein levels and inhibited adipogenesis in calreticulin-deficient ESCs, causing them to behave like the low adipogenic, wild-type ESCs. Thus, the adipogenic potential of ESCs is proportional to their calmodulin/CaMKII activity but is inversely related to their focal adhesion protein levels and degree of adhesiveness/spreading.