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TMEM16A contributes to Ca 2+ ‐activated Cl − secretion in the renal collecting duct
Author(s) -
Svenningsen Per
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.608.16
Subject(s) - purinergic receptor , secretion , extracellular , gene knockdown , microbiology and biotechnology , chemistry , small hairpin rna , cell culture , medicine , biology , biochemistry , gene , genetics
Extracellular ATP has been shown to activate purinergic signaling in epithelial cells and thereby regulate electrogenic ion transport. In the mouse renal collecting duct cell line M‐1, we found that extracellular ATP (100 nM) elicited Ca 2+ ‐activated Cl − secretion, which was blocked with the purinergic P2‐receptor antagonist suramin and the Cl − inhibitor DPC; however, the channel responsible for the Cl − is unknown. Recently, TMEM16a has been identified as a Ca 2+ ‐activated Cl − channel. We found that TMEM16a was expressed in M‐1 cells at the mRNA and protein level. We generated a M‐1 cell line with stable expression of a Dox‐inducible shRNA against TMEM16a. Induction of shRNA expression led to a knockdown of TMEM16a mRNA and protein. Knockdown of TMEM16a expression had no significant effect on ATP induced Ca 2+ in M‐1 cells; however, the Ca 2+ ‐activated Cl − secretion was significantly inhibited. Moreover, we found TMEM16a to be expressed at the apical membrane in the collecting duct of human and mouse kidneys. Taken together, the data suggest that TMEM16a is expressed in the renal collecting duct and is involved in Ca 2+ ‐activated Cl − secretion in human and mouse kidneys.