title: Anoctamin-1/TMEM16A is the major apical iodide channel of the thyrocyte.
abstract: Iodide is captured by thyrocytes through the Na(+)/I(-) symporter (NIS) before being released into the follicular lumen, where it is oxidized and incorporated into thyroglobulin for the production of thyroid hormones. Several reports point to pendrin as a candidate protein for iodide export from thyroid cells into the follicular lumen. Here, we show that a recently discovered Ca(2+)-activated anion channel, TMEM16A or anoctamin-1 (ANO1), also exports iodide from rat thyroid cell lines and from HEK 293T cells expressing human NIS and ANO1. The Ano1 mRNA is expressed in PCCl3 and FRTL-5 rat thyroid cell lines, and this expression is stimulated by thyrotropin (TSH) in rat in vivo, leading to the accumulation of the ANO1 protein at the apical membrane of thyroid follicles. Moreover, ANO1 properties, i.e., activation by intracellular calcium (i.e., by ionomycin or by ATP), low but positive affinity for pertechnetate, and nonrequirement for chloride, better fit with the iodide release characteristics of PCCl3 and FRTL-5 rat thyroid cell lines than the dissimilar properties of pendrin. Most importantly, iodide release by PCCl3 and FRTL-5 cells is efficiently blocked by T16Ainh-A01, an ANO1-specific inhibitor, and upon ANO1 knockdown by RNA interference. Finally, we show that the T16Ainh-A01 inhibitor efficiently blocks ATP-induced iodide efflux from in vitro-cultured human thyrocytes. In conclusion, our data strongly suggest that ANO1 is responsible for most of the iodide efflux across the apical membrane of thyroid cells.
authors.affiliation: Laboratoire de Biologie Moléculaire du Gène, Faculté des Sciences, Université libre de Bruxelles (ULB), Brussels, Belgium; Center for Microscopy and Molecular Imaging, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: L
authors.initials: L
authors.lastname: Twyffels
authors.affiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: A
authors.initials: A
authors.lastname: Strickaert
authors.affiliation: Laboratoire de Physiologie Moléculaire et Cellulaire, Faculté de Médecine, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: M
authors.initials: M
authors.lastname: Virreira
authors.affiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: C
authors.initials: C
authors.lastname: Massart
authors.affiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: J
authors.initials: J
authors.lastname: Van Sande
authors.affiliation: Laboratoire de Biologie Moléculaire du Gène, Faculté des Sciences, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: C
authors.initials: C
authors.lastname: Wauquier
authors.affiliation: Laboratoire de Physiologie Moléculaire et Cellulaire, Faculté de Médecine, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: R
authors.initials: R
authors.lastname: Beauwens
authors.affiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire, Université libre de Bruxelles (ULB), Brussels, Belgium; jedumont@ulb.ac.be.
authors.forename: J E
authors.initials: JE
authors.lastname: Dumont
authors.affiliation: Laboratory of Molecular Genetics, Istituto Giannina Gaslini, Genoa, Italy.
authors.forename: L J
authors.initials: LJ
authors.lastname: Galietta
authors.affiliation: Laboratoire de Physiologie Moléculaire et Cellulaire, Faculté de Médecine, Université libre de Bruxelles (ULB), Brussels, Belgium; Laboratoire d'Histologie, Histopathologie et Neuroanatomie, Faculté de Médecine, Université libre de Bruxelles (ULB), Brussels, Belgium; and.
authors.forename: A
authors.initials: A
authors.lastname: Boom
authors.affiliation: Laboratoire de Biologie Moléculaire du Gène, Faculté des Sciences, Université libre de Bruxelles (ULB), Brussels, Belgium; Center for Microscopy and Molecular Imaging, Université libre de Bruxelles (ULB), Brussels, Belgium;
authors.forename: V
authors.initials: V
authors.lastname: Kruys
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000075369:Anoctamin-1; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism; D016764:Cell Polarity; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D007454:Iodides / Q000378:metabolism*; D049990:Membrane Transport Modulators / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D051381:Rats; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013972:Thyrotropin / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: C476213:ANO1 protein, human; C552942:ANO1 protein, rat; D000075369:Anoctamin-1; D018118:Chloride Channels; D007454:Iodides; D049990:Membrane Transport Modulators; D009363:Neoplasm Proteins; D000255:Adenosine Triphosphate; D013972:Thyrotropin; D002118:Calcium
keywords: TMEM16A; anoctamin-1; iodide release; pendrin; thyrocyte