title: Exposure of colonic epithelial cells to oxidative and endoplasmic reticulum stress causes rapid potassium efflux and calcium influx.
abstract: Endoplasmic reticulum (ER) stress and oxidative stress have recently been linked to the pathogenesis of inflammatory bowel diseases. Under physiological conditions, intestinal epithelial cells are exposed to ER and oxidative stress affecting the cellular ionic homeostasis. However, these altered ion flux 'signatures' during these stress conditions are poorly characterized. We investigated the kinetics of K(+) , Ca(2+) and H(+) ion fluxes during ER and oxidative stress in a colonic epithelial cell line LS174T using a non-invasive microelectrode ion flux estimation technique. ER and oxidative stress were induced by cell exposure to tunicamycin (TM) and copper ascorbate (CuAsc), respectively, from 1 to 24 h. Dramatic K(+) efflux was observed following acute ER stress with peak K(+) efflux being -30·6 and -138·7 nmolm(-2)  s(-1) for 10 and 50 µg ml(-1) , respectively (p < 0·01). TM-dependent Ca(2+) uptake was more prolonged with peak values of 0·85 and 2·68 nmol m(-2)  s(-1) for 10 and 50 µg ml(-1) TM, respectively (p < 0·02). Ion homeostasis was also affected by the duration of ER stress. Increased duration of TM treatment from 0 to 18 h led to increases in both K(+) efflux and Ca(2+) uptake. While K(+) changes were significantly higher at each time point tested, Ca(2+) uptake was significantly higher only after prolonged treatment (18 h). CuAsc also led to an increased K(+) efflux and Ca(2+) uptake. Functional assays to investigate the effect of inhibiting K(+) efflux with tetraethylammonium resulted in increased cell viability. We conclude that ER/oxidative stress in colonic epithelial cells cause dramatic K(+) , Ca(2+) and H(+) ion flux changes, which may predispose this lineage to poor stress recovery reminiscent of that seen in inflammatory bowel diseases.
authors.affiliation: School of Agricultural Sciences, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Lana
authors.initials: L
authors.lastname: Shabala
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Walker
authors.forename: Annelie
authors.initials: A
authors.lastname: Eklund
authors.forename: Sarron
authors.initials: S
authors.lastname: Randall-Demllo
authors.forename: Sergey
authors.initials: S
authors.lastname: Shabala
authors.forename: Nuri
authors.initials: N
authors.lastname: Guven
authors.forename: Anthony L
authors.initials: AL
authors.lastname: Cook
authors.forename: Rajaraman D
authors.initials: RD
authors.lastname: Eri
mesh_terms: D002118:Calcium / Q000378:metabolism*; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D059865:Endoplasmic Reticulum Stress; D004847:Epithelial Cells / Q000378:metabolism*; D006031:Glycosylation; D006706:Homeostasis; D006801:Humans; D018384:Oxidative Stress; D011188:Potassium / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D011188:Potassium; D002118:Calcium
keywords: ER stress; colonic epithelial cells; inflammatory bowel disease; ion flux; oxidative stress