title: Dual regulation of the native ClC-K2 chloride channel in the distal nephron by voltage and pH.
abstract: ClC-K2, a member of the ClC family of Cl(-) channels and transporters, forms the major basolateral Cl(-) conductance in distal nephron epithelial cells and therefore plays a central role in renal Cl(-) absorption. However, its regulation remains largely unknown because of the fact that recombinant ClC-K2 has not yet been studied at the single-channel level. In the present study, we investigate the effects of voltage, pH, Cl(-), and Ca(2+) on native ClC-K2 in the basolateral membrane of intercalated cells from the mouse connecting tubule. The ∼10-pS channel shows a steep voltage dependence such that channel activity increases with membrane depolarization. Intracellular pH (pHi) and extracellular pH (pHo) differentially modulate the voltage dependence curve: alkaline pHi flattens the curve by causing an increase in activity at negative voltages, whereas alkaline pHo shifts the curve toward negative voltages. In addition, pHi, pHo, and extracellular Ca(2+) strongly increase activity, mainly because of an increase in the number of active channels with a comparatively minor effect on channel open probability. Furthermore, voltage alters both the number of active channels and their open probability, whereas intracellular Cl(-) has little influence. We propose that changes in the number of active channels correspond to them entering or leaving an inactivated state, whereas modulation of open probability corresponds to common gating by these channels. We suggest that pH, through the combined effects of pHi and pHo on ClC-K2, might be a key regulator of NaCl absorption and Cl(-)/HCO3 (-) exchange in type B intercalated cells.
authors: Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 06, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Institut National de la Santé et de la Recherche Médicale, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Université Paris Descartes, Sorbonne Paris Cité, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Centre National de la Recherche Scientifique ERL 8228, Centre de Recherche des Cordeliers, F-75006 Paris, France.
authors: Laurent
authors: L
authors: Pinelli
authors: Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 06, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Institut National de la Santé et de la Recherche Médicale, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Université Paris Descartes, Sorbonne Paris Cité, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Centre National de la Recherche Scientifique ERL 8228, Centre de Recherche des Cordeliers, F-75006 Paris, France.
authors: Antoine
authors: A
authors: Nissant
authors: Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 06, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Institut National de la Santé et de la Recherche Médicale, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Université Paris Descartes, Sorbonne Paris Cité, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Centre National de la Recherche Scientifique ERL 8228, Centre de Recherche des Cordeliers, F-75006 Paris, France.
authors: Aurélie
authors: A
authors: Edwards
authors: Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 06, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Institut National de la Santé et de la Recherche Médicale, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Université Paris Descartes, Sorbonne Paris Cité, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Centre National de la Recherche Scientifique ERL 8228, Centre de Recherche des Cordeliers, F-75006 Paris, France.
authors: Stéphane
authors: 0000-0002-3626-7650
authors: S
authors: Lourdel
authors: Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 06, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Institut National de la Santé et de la Recherche Médicale, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Université Paris Descartes, Sorbonne Paris Cité, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Centre National de la Recherche Scientifique ERL 8228, Centre de Recherche des Cordeliers, F-75006 Paris, France jacques.teulon@upmc.fr.
authors: Jacques
authors: J
authors: Teulon
authors: Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 06, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Institut National de la Santé et de la Recherche Médicale, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Université Paris Descartes, Sorbonne Paris Cité, UMR_S 1138, Centre de Recherche des Cordeliers, F-75006 Paris, France Centre National de la Recherche Scientifique ERL 8228, Centre de Recherche des Cordeliers, F-75006 Paris, France.
authors: Marc
authors: M
authors: Paulais
mesh_terms: D000818:Animals; D027321:Anion Transport Proteins / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D018118:Chloride Channels / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D015640:Ion Channel Gating / Q000502:physiology; D008297:Male; D051379:Mice; D009399:Nephrons / Q000378:metabolism*; D012965:Sodium Chloride / Q000378:metabolism
chemical_list: D027321:Anion Transport Proteins; D018118:Chloride Channels; D002712:Chlorides; C486573:Clcnkb protein, mouse; D012965:Sodium Chloride; D002118:Calcium