Understanding the role of ZO-1 in cell phenotype and gene expression through myosin inhibition

Abstract

Zonula occludens-1 (ZO-1) is a tight junction protein contributing to junctional formation as well as mediating interactions with junctional proteins and the actomyosin cytoskeleton. To investigate the role of ZO-1, Madin-Darby canine kidney (MDCK) II knockout cells lacking ZO-1 were created utilizing a CRISPR gene editing platform. These ZO-1 KO cells display characteristic phenotypic changes including actin accumulation and junction linearization. We evaluated membrane tension and found ZO-1 loss leads to increased membrane tension and nuclear density. We hypothesized that due changes in cytoskeletal organization and membrane tension the ZO-1 KO cells would spread less following plating. It is suspected that phenotypic changes related to actin accumulation may be the result of myosin dysregulation in the absence of ZO-1. ZO-1 KO cells that are exposed to blebbistatin, a myosin II inhibitor, resemble wild type cells in cell height and junction linearization. RNAseq and qRT-PCR were used to investigate gene expression changes related to ZO-1 loss to further interrogate ZO-1’s role in regulating membrane tension. This study highlights and expands our understanding of how ZO-1 regulates the mechanical properties of epithelial cells. Trinity University, Mach Research Fellowship This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.