HV1 Acts as a Sodium Sensor and Promotes Superoxide Production in Medullary Thick Ascending Limb of Dahl Salt-Sensitive Rats Chunhua Jin, Jingping Sun, Carly A. Stilphen, Susan M.E. Smith, Hiram Ocasio, Brent Bermingham, Sandip Darji, Avirup Guha, Roshan Patel, Aron M. Geurts, Howard J. Jacob, Nevin A. Lambert and Paul M. O'Connor Hypertension. published online June 16, 2014; Hypertension is published by the American Heart Association, 7272 Greenville Avenue, Dallas, TX 75231 Copyright © 2014 American Heart Association, Inc. All rights reserved. Print ISSN: 0194-911X. Online ISSN: 1524-4563
The online version of this article, along with updated information and services, is located on the World Wide Web at: http://hyper.ahajournals.org/content/early/2014/06/16/HYPERTENSIONAHA.114.03549
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Original Article HV1 Acts as a Sodium Sensor and Promotes Superoxide Production in Medullary Thick Ascending Limb of Dahl Salt-Sensitive Rats Chunhua Jin, Jingping Sun, Carly A. Stilphen, Susan M.E. Smith, Hiram Ocasio, Brent Bermingham, Sandip Darji, Avirup Guha, Roshan Patel, Aron M. Geurts, Howard J. Jacob, Nevin A. Lambert, Paul M. O’Connor Abstract—We previously characterized a H+ transport pathway in medullary thick ascending limb nephron segments that when activated stimulated the production of superoxide by nicotinamide adenine dinucleotide phosphate oxidase. Importantly, the activity of this pathway was greater in Dahl salt-sensitive rats than salt-resistant (SS.13BN) rats, and superoxide production was enhanced in low Na+ media. The goal of this study was to determine the molecular identity of this pathway and its relationship to Na+. We hypothesized that the voltage-gated proton channel, HV1, was the source of superoxide-stimulating H+ currents. To test this hypothesis, we developed HV1−/− null mutant rats on the Dahl saltsensitive rat genetic background using zinc-finger nuclease gene targeting. HV1 could be detected in medullary thick limb from wild-type rats. Intracellular acidification using an NH4Cl prepulse in 0 sodium/BaCl2 containing media resulted in superoxide production in thick limb from wild-type but not HV1−/− rats (P
The online version of this article, along with updated information and services, is located on the World Wide Web at: http://hyper.ahajournals.org/content/early/2014/06/16/HYPERTENSIONAHA.114.03549
Data Supplement (unedited) at: http://hyper.ahajournals.org/content/suppl/2014/06/16/HYPERTENSIONAHA.114.03549.DC1.html
Permissions: Requests for permissions to reproduce figures, tables, or portions of articles originally published in Hypertension can be obtained via RightsLink, a service of the Copyright Clearance Center, not the Editorial Office. Once the online version of the published article for which permission is being requested is located, click Request Permissions in the middle column of the Web page under Services. Further information about this process is available in the Permissions and Rights Question and Answer document. Reprints: Information about reprints can be found online at: http://www.lww.com/reprints Subscriptions: Information about subscribing to Hypertension is online at: http://hyper.ahajournals.org//subscriptions/
Downloaded from http://hyper.ahajournals.org/ at Queen Mary, University of London on July 3, 2014
Original Article HV1 Acts as a Sodium Sensor and Promotes Superoxide Production in Medullary Thick Ascending Limb of Dahl Salt-Sensitive Rats Chunhua Jin, Jingping Sun, Carly A. Stilphen, Susan M.E. Smith, Hiram Ocasio, Brent Bermingham, Sandip Darji, Avirup Guha, Roshan Patel, Aron M. Geurts, Howard J. Jacob, Nevin A. Lambert, Paul M. O’Connor Abstract—We previously characterized a H+ transport pathway in medullary thick ascending limb nephron segments that when activated stimulated the production of superoxide by nicotinamide adenine dinucleotide phosphate oxidase. Importantly, the activity of this pathway was greater in Dahl salt-sensitive rats than salt-resistant (SS.13BN) rats, and superoxide production was enhanced in low Na+ media. The goal of this study was to determine the molecular identity of this pathway and its relationship to Na+. We hypothesized that the voltage-gated proton channel, HV1, was the source of superoxide-stimulating H+ currents. To test this hypothesis, we developed HV1−/− null mutant rats on the Dahl saltsensitive rat genetic background using zinc-finger nuclease gene targeting. HV1 could be detected in medullary thick limb from wild-type rats. Intracellular acidification using an NH4Cl prepulse in 0 sodium/BaCl2 containing media resulted in superoxide production in thick limb from wild-type but not HV1−/− rats (P
