Apparent intermediate K conductance channel hyposmotic activation in human lens epithelial cells

American Journal of Physiology. Cell Physiology
P K LaufNorma C Adragna

Abstract

This study explores the nature of K fluxes in human lens epithelial cells (LECs) in hyposmotic solutions. Total ion fluxes, Na-K pump, Cl-dependent Na-K-2Cl (NKCC), K-Cl (KCC) cotransport, and K channels were determined by 85Rb uptake and cell K (Kc) by atomic absorption spectrophotometry, and cell water gravimetrically after exposure to ouabain +/- bumetanide (Na-K pump and NKCC inhibitors), and ion channel inhibitors in varying osmolalities with Na, K, or methyl-d-glucamine and Cl, sulfamate, or nitrate. Reverse transcriptase polymerase chain reaction (RT-PCR), Western blot analyses, and immunochemistry were also performed. In isosmotic (300 mosM) media approximately 90% of the total Rb influx occurred through the Na-K pump and NKCC and approximately 10% through KCC and a residual leak. Hyposmotic media (150 mosM) decreased K(c) by a 16-fold higher K permeability and cell water, but failed to inactivate NKCC and activate KCC. Sucrose replacement or extracellular K to >57 mM, but not Rb or Cs, in hyposmotic media prevented Kc and water loss. Rb influx equaled Kc loss, both blocked by clotrimazole (IC50 approximately 25 microM) and partially by 1-[(2-chlorophenyl) diphenylmethyl]-1H-pyrazole (TRAM-34) inhibitors of the IK chann...Continue Reading

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Citations

Aug 7, 2012·Pflügers Archiv : European journal of physiology·Silvia Cruz-RangelHerminia Pasantes-Morales
May 7, 2010·American Journal of Physiology. Cell Physiology·Christian BarmeyerVazhaikkurichi M Rajendran
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Jan 17, 2019·American Journal of Physiology. Cell Physiology·Peter K LaufNorma C Adragna
Jan 6, 2010·Journal of Cellular Physiology·Ameet A ChimotePeter K Lauf
Jan 8, 2009·Physiological Reviews·Else K HoffmannStine F Pedersen
Oct 16, 2014·American Journal of Physiology. Cell Physiology·Peter K LaufNorma C Adragna

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