Lack of coupling between membrane stretching and pannexin-1 hemichannels
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We investigated whether pannexin-1, a carbenoxolone-sensitive hemichannel activated in erythrocytes by swelling, could be activated by swelling stress and contribute to swelling-activated chloride currents (ICl,swell) in HEK-293 cells. We used ethidium bromide uptake as an index of pannexin-1 activation and IC,swell activation as an index of plasma membrane stretching. ICl,swell activated by a hypotonic solution was reversible inhibited by carbenoxolone (IC50 98 ± 5 μM). However, the hypotonic solution that activated ICl,swell did not induce ethidium bromide uptake indicating that pannexin-1 was not activated by cell swelling. The mimetic peptide 10panx1, a pannexin-1 antagonist, did not affect ICl,swell activation but completely inhibited the ATP-induced ethidium bromide uptake coupled to P2X7 receptors activation. We conclude that carbenoxolone directly inhibited ICl,swell independent of pannexin-1 and that pannexin-1 hemichannels are not activated by swelling in HEK-293 cells. © 2009 Elsevier Inc. All rights reserved.
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ATP release; Carbenoxolone; Cell swelling; Dye uptake; Pannexin-1; Swelling-activated chloride current adenosine triphosphate; carbenoxolone; ethidium bromide; hypotonic solution; pannexin 1; purinergic P2X7 receptor; unclassified drug; article; binding site; cell membrane; cell swelling; chloride current; controlled study; erythrocyte; human; human cell; priority journal; protein function; Biological Transport; Carbenoxolone; Cell Line; Cell Membrane; Connexins; Ethidium; Humans; Membrane Potentials; Nerve Tissue Proteins; Osmosis; Stress, Mechanical
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