Generation and functional characterization of epithelial cells with stable expression of SLC26A9 Cl- channels

American Journal of Physiology. Lung Cellular and Molecular Physiology
Johanna J SalomonM Mall

Abstract

Recent studies identified the SLC26A9 Cl(-) channel as a modifier and potential therapeutic target in cystic fibrosis (CF). However, understanding of the regulation of SLC26A9 in epithelia remains limited and cellular models with stable expression for biochemical and functional studies are missing. We, therefore, generated Fisher rat thyroid (FRT) epithelial cells with stable expression of HA-tagged SLC26A9 via retroviral transfection and characterized SLC26A9 expression and function using Western blotting, immunolocalization, whole cell patch-clamp, and transepithelial bioelectric studies in Ussing chambers. We demonstrate stable expression of SLC26A9 in transfected FRT (SLC26A9-FRT) cells on the mRNA and protein level. Immunolocalization and Western blotting detected SLC26A9 in different intracellular compartments and to a lesser extent at the cell surface. Whole cell patch-clamp recordings demonstrated significantly increased constitutive Cl(-) currents in SLC26A9-FRT compared with control-transduced FRT (Control-FRT) cells (P < 0.01). Similar, transepithelial measurements showed that the basal short circuit current was significantly increased in SLC26A9-FRT vs. Control-FRT cell monolayers (P < 0.01). SLC26A9-mediated Cl(-) ...Continue Reading

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Citations

Apr 30, 2016·Pflügers Archiv : European journal of physiology·Meng WuSeth L Alper
Jan 22, 2017·Cell and Tissue Research·Zhe Zhou-SuckowMarcus A Mall
Jul 28, 2016·American Journal of Physiology. Lung Cellular and Molecular Physiology·Jie JiaAli Önder Yildirim
Apr 1, 2017·American Journal of Physiology. Lung Cellular and Molecular Physiology·Carol A BertrandRaymond A Frizzell
Nov 16, 2018·Annals of the American Thoracic Society·Marcus A MallRichard C Boucher
Oct 4, 2019·The Journal of Clinical Investigation·Anh-Thu N LamGarry R Cutting
Oct 14, 2017·American Journal of Physiology. Lung Cellular and Molecular Physiology·Rafal BartoszewskiJames F Collawn
Aug 5, 2017·American Journal of Physiology. Lung Cellular and Molecular Physiology·James David LondinoSadis Matalon
Jul 31, 2018·Human Molecular Genetics·Lisa J StrugAnn Harris
Nov 26, 2020·American Journal of Physiology. Lung Cellular and Molecular Physiology·Chloé GrebertClarisse Vandebrouck
Aug 4, 2020·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Onofrio LaselvaChristine E Bear
Feb 21, 2021·The Journal of Allergy and Clinical Immunology·Johanna J SalomonMarcus A Mall
Apr 4, 2021·International Journal of Molecular Sciences·Miroslaw ZajacIsabelle Sermet-Gaudelus
Jul 31, 2021·Journal of Experimental Pharmacology·Madalena C PintoMiquéias Lopes-Pacheco
Oct 21, 2021·American Journal of Physiology. Lung Cellular and Molecular Physiology·Mads B LarsenCarol A Bertrand

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