Chloride Channel ClC- 2 Enhances Intestinal Epithelial Tight Junction Barrier Function Via Regulation of Caveolin-1 and Caveolar Trafficking of Occludin

Overview

Journal Exp Cell Res

Specialty Cell Biology

Date 2017 Feb 6

PMID 28161538

Citations 23

Authors

Prashant K Nighot

Lana Leung

Thomas Y Ma

Affiliations

Soon will be listed here.

Abstract

Previous studies have demonstrated that the chloride channel ClC-2 plays a critical role in intestinal epithelial tight junction (TJ) barrier function via intracellular trafficking of TJ protein occludin. To study the mechanism of ClC-2-mediated TJ barrier function and intracellular trafficking of occludin, we established ClC-2 over-expressing Caco-2 cell line (Caco-2) by full length ClC-2 ORF transfection. ClC-2 over-expression (Caco-2) significantly enhanced TJ barrier (increased TER by ≥2 times and reduced inulin flux by 50%) compared to control Caco-2 cells. ClC-2 over-expression (Caco-2) increased occludin protein level compared to control Caco-2 cells. Surface biotinylation assay revealed reduced steady state endocytosis of occludin in Caco-2 cells. Furthermore, ClC-2 over-expression led to reduction in caveolin-1 protein level and diminishment of caveolae assembly. Caveolae disruption increased TJ permeability in control but not ClC-2 over-expressing Caco-2 cells. Selective ClC-2 channel blocker GaTx2 caused an increase in caveolin-1 protein level and reduced occludin level. Delivery of cell permeable caveolin-1 scaffolding domain reduced the occludin protein level. Over all, these results suggest that ClC- 2 enhances TJ barrier function in intestinal epithelial cells via regulation of caveolin-1 and caveolae-mediated trafficking of occludin.

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References

Nighot M, Nighot P, Ma T, Malinowska D, Shull G, Cuppoletti J . Genetic Ablation of the ClC-2 Cl- Channel Disrupts Mouse Gastric Parietal Cell Acid Secretion. PLoS One. 2015; 10(9):e0138174. PMC: 4574764. DOI: 10.1371/journal.pone.0138174. View

Collins B, Davis M, Hancock J, Parton R . Structure-based reassessment of the caveolin signaling model: do caveolae regulate signaling through caveolin-protein interactions?. Dev Cell. 2012; 23(1):11-20. PMC: 3427029. DOI: 10.1016/j.devcel.2012.06.012. View

Weiss C, Guan Q, Ma Y, Qing G, Bernstein C, Warrington R . The potential protective role of caveolin-1 in intestinal inflammation in TNBS-induced murine colitis. PLoS One. 2015; 10(3):e0119004. PMC: 4355071. DOI: 10.1371/journal.pone.0119004. View

Mir H, Meena A, Chaudhry K, Shukla P, Gangwar R, Manda B . Occludin deficiency promotes ethanol-induced disruption of colonic epithelial junctions, gut barrier dysfunction and liver damage in mice. Biochim Biophys Acta. 2016; 1860(4):765-74. PMC: 4776745. DOI: 10.1016/j.bbagen.2015.12.013. View

Turner J . Intestinal mucosal barrier function in health and disease. Nat Rev Immunol. 2009; 9(11):799-809. DOI: 10.1038/nri2653. View

Nusrat A, Parkos C, Verkade P, Foley C, Liang T, Eastburn K . Tight junctions are membrane microdomains. J Cell Sci. 2000; 113 ( Pt 10):1771-81. DOI: 10.1242/jcs.113.10.1771. View

Wong V, Gumbiner B . A synthetic peptide corresponding to the extracellular domain of occludin perturbs the tight junction permeability barrier. J Cell Biol. 1997; 136(2):399-409. PMC: 2134825. DOI: 10.1083/jcb.136.2.399. View

Nighot P, Blikslager A . Chloride channel ClC-2 modulates tight junction barrier function via intracellular trafficking of occludin. Am J Physiol Cell Physiol. 2011; 302(1):C178-87. DOI: 10.1152/ajpcell.00072.2011. View

Moeser A, Haskell M, Shifflett D, Little D, Schultz B, Blikslager A . ClC-2 chloride secretion mediates prostaglandin-induced recovery of barrier function in ischemia-injured porcine ileum. Gastroenterology. 2004; 127(3):802-15. DOI: 10.1053/j.gastro.2004.06.004. View

10.

Casas E, Barron C, Francis S, McCormack J, McCarthy K, Schneeberger E . Cholesterol efflux stimulates metalloproteinase-mediated cleavage of occludin and release of extracellular membrane particles containing its C-terminal fragments. Exp Cell Res. 2009; 316(3):353-65. PMC: 2812682. DOI: 10.1016/j.yexcr.2009.10.020. View

11.

Parton R . Caveolae meet endosomes: a stable relationship?. Dev Cell. 2004; 7(4):458-60. DOI: 10.1016/j.devcel.2004.09.009. View

12.

Francis S, Kelly J, McCormack J, Rogers R, Lai J, Schneeberger E . Rapid reduction of MDCK cell cholesterol by methyl-beta-cyclodextrin alters steady state transepithelial electrical resistance. Eur J Cell Biol. 1999; 78(7):473-84. DOI: 10.1016/s0171-9335(99)80074-0. View

13.

McCarthy K, Skare I, Stankewich M, Furuse M, Tsukita S, Rogers R . Occludin is a functional component of the tight junction. J Cell Sci. 1996; 109 ( Pt 9):2287-98. DOI: 10.1242/jcs.109.9.2287. View

14.

Mandel L, Bacallao R, Zampighi G . Uncoupling of the molecular 'fence' and paracellular 'gate' functions in epithelial tight junctions. Nature. 1993; 361(6412):552-5. DOI: 10.1038/361552a0. View

15.

Bosl M, Stein V, Hubner C, Zdebik A, Jordt S, Mukhopadhyay A . Male germ cells and photoreceptors, both dependent on close cell-cell interactions, degenerate upon ClC-2 Cl(-) channel disruption. EMBO J. 2001; 20(6):1289-99. PMC: 145530. DOI: 10.1093/emboj/20.6.1289. View

16.

Nighot P, Moeser A, Ryan K, Ghashghaei T, Blikslager A . ClC-2 is required for rapid restoration of epithelial tight junctions in ischemic-injured murine jejunum. Exp Cell Res. 2008; 315(1):110-8. DOI: 10.1016/j.yexcr.2008.10.001. View

17.

Mruk D, Lau A, Conway A . Crosstalk between Rab GTPases and cell junctions. Contraception. 2005; 72(4):280-90. DOI: 10.1016/j.contraception.2005.03.013. View

18.

Xu S, Xue X, You K, Fu J . Caveolin-1 regulates the expression of tight junction proteins during hyperoxia-induced pulmonary epithelial barrier breakdown. Respir Res. 2016; 17(1):50. PMC: 4866358. DOI: 10.1186/s12931-016-0364-1. View

19.

Xie L, Xue X, Taylor M, Ramakrishnan S, Nagaoka K, Hao C . Hypoxia-inducible factor/MAZ-dependent induction of caveolin-1 regulates colon permeability through suppression of occludin, leading to hypoxia-induced inflammation. Mol Cell Biol. 2014; 34(16):3013-23. PMC: 4135598. DOI: 10.1128/MCB.00324-14. View

20.

Xia W, Wong E, Mruk D, Cheng C . TGF-beta3 and TNFalpha perturb blood-testis barrier (BTB) dynamics by accelerating the clathrin-mediated endocytosis of integral membrane proteins: a new concept of BTB regulation during spermatogenesis. Dev Biol. 2008; 327(1):48-61. PMC: 2758298. DOI: 10.1016/j.ydbio.2008.11.028. View