Regulation of ClC-K/barttin by endocytosis influences distal convoluted tubule hyperplasia
Por:
Mayayo-Vallverdú C, Gaitán-Peñas H, Armand-Ugon M, Muhaisen A, Prat E, Castellanos A, Elorza-Vidal X, de Heredia ML, Alonso-Gardón M, Pérez-Rius C, Vecino-Pérez M, Mallen A, Errasti-Murugarren E, Hueso M, Artuch-Iriberri R, Nunes V and Estévez R
Publicada:
1 sep 2024
Ahead of Print:
1 ago 2024
Resumen:
ClC-K/barttin channels are involved in the transepithelial transport of chloride in the kidney and inner ear. Their physiological role is crucial in humans because mutations in CLCNKB or BSND, encoding ClC-Kb and barttin, cause Bartter's syndrome types III and IV, respectively. In vitro experiments have shown that an amino acid change in a proline-tyrosine motif in the C-terminus of barttin stimulates ClC-K currents. The molecular mechanism of this enhancement and whether this potentiation has any in vivo relevance remains unknown. We performed electrophysiological and biochemical experiments in Xenopus oocytes and kidney cells co-expressing ClC-K and barttin constructs. We demonstrated that barttin possesses a Yxx & Oslash; motif and, when mutated, increases ClC-K plasma membrane stability, resulting in larger currents. To address the impact of mutating this motif in kidney physiology, we generated a knock-in mouse. Comparing wild-type (WT) and knock-in mice under a standard diet, we could not observe any difference in ClC-K and barttin protein levels or localization, either in urinary or plasma parameters. However, under a high-sodium low-potassium diet, known to induce hyperplasia of distal convoluted tubules, knock-in mice exhibit reduced hyperplasia compared to WT mice. In summary, our in vitro and in vivo studies demonstrate that the previously identified PY motif is indeed an endocytic Yxx & Oslash; motif in which mutations cause a gain of function of the channel.
Filiaciones:
Mayayo-Vallverdú C:
Genes, Disease and Therapy Program, Molecular Genetics Laboratory-IDIBELL, Genetics Section, Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
Gaitán-Peñas H:
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Armand-Ugon M:
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Muhaisen A:
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Prat E:
Genes, Disease and Therapy Program, Molecular Genetics Laboratory-IDIBELL, Genetics Section, Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
Castellanos A:
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
Elorza-Vidal X:
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
de Heredia ML:
Genes, Disease and Therapy Program, Molecular Genetics Laboratory-IDIBELL, Genetics Section, Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
Alonso-Gardón M:
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Pérez-Rius C:
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Vecino-Pérez M:
Genes, Disease and Therapy Program, Molecular Genetics Laboratory-IDIBELL, Genetics Section, Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
Mallen A:
Department of Nephrology, Hospital Universitart Bellvitge and Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Spain
Errasti-Murugarren E:
Genes, Disease and Therapy Program, Molecular Genetics Laboratory-IDIBELL, Genetics Section, Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
Hueso M:
Department of Nephrology, Hospital Universitart Bellvitge and Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Spain
Artuch-Iriberri R:
Institut de Recerca Sant Joan de Déu, Barcelona, Spain
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
Nunes V:
Genes, Disease and Therapy Program, Molecular Genetics Laboratory-IDIBELL, Genetics Section, Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
Estévez R:
Physiology Unit, Department of Physiological Sciences, School of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat, Spain
Genes, Disease and Therapy Program, Physiology and pathology of the functional relationship between glia and neurons-IDIBELL, L'Hospitalet de Llobregat, Spain
Centro de Investigación en red de enfermedades raras (CIBERER), Instituto de Salud Carlos III, Madrid, Spain
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