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TMEM16A confers receptor-activated calcium-dependent chloride conductance

Abstract

Calcium (Ca2+)-activated chloride channels are fundamental mediators in numerous physiological processes including transepithelial secretion, cardiac and neuronal excitation, sensory transduction, smooth muscle contraction and fertilization. Despite their physiological importance, their molecular identity has remained largely unknown. Here we show that transmembrane protein 16A (TMEM16A, which we also call anoctamin 1 (ANO1)) is a bona fide Ca2+-activated chloride channel that is activated by intracellular Ca2+ and Ca2+-mobilizing stimuli. With eight putative transmembrane domains and no apparent similarity to previously characterized channels, ANO1 defines a new family of ionic channels. The biophysical properties as well as the pharmacological profile of ANO1 are in full agreement with native Ca2+-activated chloride currents. ANO1 is expressed in various secretory epithelia, the retina and sensory neurons. Furthermore, knockdown of mouse Ano1 markedly reduced native Ca2+-activated chloride currents as well as saliva production in mice. We conclude that ANO1 is a candidate Ca2+-activated chloride channel that mediates receptor-activated chloride currents in diverse physiological processes.

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Figure 1: ANO1-transfected cells respond with robust inward currents to GPCR stimulation.
Figure 2: ANO1 is a Cl - channel.
Figure 3: ANO1 is activated by intracellular Ca 2+ in a voltage-dependent manner.
Figure 4: ANO1 is expressed in transport epithelia and other tissues.
Figure 5: Mouse Ano1 siRNA reduces pilocarpine-induced salivary output.

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Acknowledgements

We thank R. MacKinnon and J. Wood for a review of themanuscript. We also thank B. Hille for his technical comment on ion permeability. This work was supported by Acceleration Research of MOEST/KOSEF of Korea. This work was also supported by the Brain Korea 21 Project and by the Wellcome Trust Foundation, UK (R.R.).

Author Contributions Y.D.Y. cloned the channel and its mutants and GPCRs. W.-S.S. worked on bioinformatics. Y.D.Y. recorded currents in oocytes. H.C., B.L. and M.H.T. recorded whole-cell currents. J.Y.K. recorded single-channel currents. J.L. and Y.C. worked on siRNA in vivo. B.-M.K. worked on western blot. R.R. measured Ca2+ and currents. S.P.P. measured electrolytes in saliva. Y.K.S. worked on immunohistochemistry. U.O. designed and supervised experiments, and wrote the manuscript.

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Correspondence to Uhtaek Oh.

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Yang, Y., Cho, H., Koo, J. et al. TMEM16A confers receptor-activated calcium-dependent chloride conductance. Nature 455, 1210–1215 (2008). https://doi.org/10.1038/nature07313

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