Research resource: Monitoring endoplasmic reticulum membrane integrity in beta-cells at the single-cell level

Kohsuke Kanekura, Washington University School of Medicine
Jianhong Ou, University of Massachusetts Medical School
Takashi Hara, Washington University School of Medicine
Lihua Julie Zhu, University of Massachusetts Medical School
Fumihiko Urano, Washington University School of Medicine

Abstract

Endoplasmic reticulum (ER) membrane integrity is an emerging target for human chronic diseases associated with ER stress. Despite the underlying importance of compromised ER membrane integrity in disease states, the entire process leading to ER membrane permeabilization and cell death is still not clear due to technical limitations. Here we describe a novel method for monitoring ER membrane integrity at the single-cell level in real time. Using a beta-cell line expressing ER-targeted redox sensitive green fluorescent protein, we could identify a beta-cell population undergoing ER membrane permeabilization induced by palmitate and could monitor cell fate and ER stress of these cells at the single-cell level. Our method could be used to develop a novel therapeutic modality targeting the ER membrane for ER-associated disorders, including beta-cell death in diabetes, neurodegeneration, and Wolfram syndrome.