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Ano6 functions as a positive modulator of volume‐regulated anion channels

Anoctamin 6 (Ano6) is a homologue of Ano1 that functions as a calcium‐activated chloride channel. Recently, Ano6 has been identified as an essential component of outwardly rectifying chloride channels. It has been also shown that Ano6 and Ano1 are activated by hypotonic cell swelling (HTS) and might be a part of an epithelial volume‐regulated anion channel (VRAC). Using quantitative PCR, we showed that Ano6 expression levels correlated with detection of large VRAC currents in smooth muscle‐like BC3H1 and colon carcinoma Caco‐2 cells. Overexpression of Ano6 in HEK293 cells resulted in significant increase of HTS‐evoked anion currents when compared to untransfected cells. In contrast to Ano1, Ano6 was not activated by elevation of intracellular Ca2+. Moreover, co‐expression with caveolin‐1, a main component of caveolae plasma membrane and a positive regulator of VRAC, significantly increases HTS‐evoked currents in Ano6‐overexpressing HEK293 cells. Similarly, transient Ano6 expression in Caco‐2 cells generated large increase of HTS‐evoked currents when compared to the untransfected cells. Recorded currents displayed known characteristics of VRAC such as a rapid deactivation at high positive potentials and block by tamoxifen and mibefradil. Our results suggest that Ano6 is a positive modulator of VRAC function. Supported by Interuniversity Poles of Attraction Program and Program Financing, KU Leuven.