Confluence and tight junction dependence of volume regulation in epithelial tissue

Epithelial cell volume regulation is a key component to tissue stability and dynamics. In particular, how cells respond to osmotic stresses is of significant physiological interest in kidney epithelial tissue. For individual mammalian cells, it is well established that Na-K-2Cl cotransporter (NKCC) channels mediate cell volume homeostasis in response to hyperosmotic stress. However, whether mature epithelium responds similarly is not well known. Here we show that while small colonies of madin darby canine kidney (MDCK) epithelial cells behave similarly to single cells and exhibit volume homeostasis that is dependent on the NKCC channel function, mature epithelial tissue does not. Instead, the cell volume decreases by 33% when confluent monolayers or acini formed from MDCK cells are subjected to hyperosmotic stress. We show that the tight junction protein zonula occludins-1 (ZO-1), and Rho-associated kinase (ROCK) are essential for osmotic regulation of cell volume in mature epithelium. Because these both are known to be essential for tight junction assembly, this strongly suggests a role for tight junctions in changing volume response in mature epithelium. Thus, tight junctions act either directly or indirectly in osmotic pressure response of epithelial tissue to suppress volume homeostasis common to isolated epithelial cells.