Simultaneous functional expression of swelling and forskolin-activated chloride currents in primary cultures of rabbit distal convoluted tubule |
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| Institution: | 1. Division of Nephrology and Hypertension, Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee, USA;2. Vanderbilt Center for Kidney Disease, Vanderbilt University School of Medicine, Nashville, Tennessee, USA;3. Division of Nephrology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China;4. Department of Veterans Affairs, Nashville, Tennessee, USA;1. Welch Center for Prevention, Epidemiology, and Clinical Research, Baltimore, MD;2. Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD;3. Division of Nephrology, Department of Medicine, Johns Hopkins School of Medicine, Baltimore, MD;4. Division of Epidemiology & Community Health, University of Minnesota School of Public Health, Minneapolis, MN;5. William B. Schwartz Division of Nephrology, Department of Medicine, Tufts Medical Center, Boston, MA;6. Division of General Internal Medicine, Department of Medicine, Johns Hopkins School of Medicine, Baltimore, MD;1. Department of Kinesiology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1;2. School of Public Health and Health Systems, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1;1. Department of Molecular Cell Biology, Institute for Cancer Research, Oslo University Hospital-The Norwegian Radium Hospital, 0379 Oslo, Norway;2. Centre for Cancer Biomedicine, Faculty of Medicine, University of Oslo, 0379 Oslo, Norway;3. Zora Biosciences Oy, 02150 Espoo, Finland;4. Division of Radiology and Nuclear Medicine, Oslo University Hospital, 0379 Oslo, Norway;5. Department of Urology, Oslo University Hospital, 0586 Oslo, Norway;6. Department of Biosciences, University of Oslo, 0316 Oslo, Norway;1. The Farr Institute of Health Informatics Research, London, UK;2. Epidemiology and Public Health, University College London, London, UK;3. National Centre for Cardiovascular Prevention and Outcomes, Institute of Cardiovascular Science, University College London, London, UK;4. Barts and The London National Institute for Health Research Cardiovascular Biomedical Research Unit, Queen Mary University of London, London, UK;5. William Harvey Research Institute and the Barts National Institute for Health Research Biomedical Research Unit, Queen Mary University of London, London, UK;6. Epidemiology and Population Health, London School of Hygiene & Tropical Medicine, London, UK;7. University College London and National Institute for Health Research UCL Hospitals Biomedical Research Centre, London, UK;8. MRC Biostatistics Unit, Cambridge, UK |
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| Abstract: | Ionic Cl? currents induced by cell swelling and forskolin were studied in primary cultures of rabbit distal convoluted tubule (DCTb) by the whole-cell patch clamp technique. We identified a Cl? conductance activated by cell swelling with an hyperosmotic pipette solution. The initial current exhibited an outwardly rectifying I-V relationship, whereas steady state current showed strong decay at depolarized membrane potentials. The ion selectivity was I? > Br? > Cl? > > glutamate. The forskolin-activated Cl? conductance demonstrated a linear I-V relationship and its ion selectivity was Br? > Cl? > I? > glutamate. This last conductance could be related to the CFTR (cystic fibrosis transmembrane conductance regulator) previously identified in these cells. NPPB inhibited both Cl? currents, and DIDS inhibited only the swelling-activated Cl? current. Forskolin had no effect on the activation of the swelling-activated Cl? current. In DCTb cells which exhibited swelling-activated Cl? currents subsequently inhibited by DIDS, forskolin could activate CFTR related Cl? currents. In the continuous presence of I? which inhibited CFTR conductance, forskolin did not modify the swelling-activated current. The results suggest that both Cl? conductances could be co-expressed in the same DCTb cell and that CFTR did not modulate the swelling-activated conductance. |
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