title: Research resource: Monitoring endoplasmic reticulum membrane integrity in β-cells at the single-cell level.
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 β-cell line expressing ER-targeted redox sensitive green fluorescent protein, we could identify a β-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 β-cell death in diabetes, neurodegeneration, and Wolfram syndrome.
authors.affiliation: Department of Medicine (K.K., T.H., F.U.), Division of Endocrinology, Metabolism, and Lipid Research, and Department of Pathology and Immunology (F.U.), Washington University School of Medicine, St Louis, Missouri 63110; and Programs in Gene Function and Expression, Molecular Medicine, and Bioinformatics and Integrative Biology (J.O., L.J.Z.), University of Massachusetts Medical School, Worcester, Massachusetts 01655.
authors.forename: Kohsuke
authors.initials: K
authors.lastname: Kanekura
authors.forename: Jianhong
authors.initials: J
authors.lastname: Ou
authors.forename: Takashi
authors.initials: T
authors.lastname: Hara
authors.forename: Lihua J
authors.initials: LJ
authors.lastname: Zhu
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Urano
mesh_terms: D000818:Animals; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism*; D008563:Membrane Lipids / Q000737:chemistry; D051379:Mice; D010168:Palmitates / Q000494:pharmacology; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D008563:Membrane Lipids; D010168:Palmitates; D049452:Green Fluorescent Proteins