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The proton channel HV1 is expressed in the medullary thick ascending limb of the kidney (1134.1)
Author(s) -
Stilphen Carly,
Sun Jingping,
Lambert Nevin,
O’Connor Paul
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1134.1
Subject(s) - endocrinology , medicine , western blot , kidney , renal medulla , superoxide , messenger rna , northern blot , chemistry , biology , microbiology and biotechnology , gene , biochemistry , enzyme
Oxidative stress in the renal outer‐medulla contributes to the development of hypertension and renal disease in Dahl salt‐sensitive (SS) rats. We have previously demonstrated that an unknown proton channel is responsible for enhanced superoxide production in medullary thick ascending limb (mTAL) of SS rats. We hypothesized that HV1 is responsible for superoxide producing H + currents and is expressed in mTAL of SS rats. In order to test our hypothesis we quantitated HV1 mRNA (RT‐PCR) and protein (immunohistochemistry) specifically within mTAL of wild‐type SS rats and in a HV1 ‐/‐ strain of Dahl SS rats in which we mutated the HV1 gene using a Zn‐finger nuclease. We confirmed complete loss of Hv1 protein in HV1 ‐/‐ rats both by whole‐cell voltage clamp of peritoneal macrophages and by Western blot/anti‐HV1 staining in spleen (positive control tissue). Using the same antibody, HV1 protein expression was found to be significantly greater in mTAL of wild‐type rats compared to presumed background in HV1 ‐/‐ rats (4382 ± 921 vs 1459 ± 708AU respectively, n=5/p=0.036). In addition, mRNA for HV1 was expressed 6.4 fold higher in mTAL of wild‐type rats (n=4/p=.04). We conclude that HV1 is expressed in the mTAL of Dahl SS rats. Our data indicates that HV1 could potentially be responsible for superoxide producing H + currents in mTAL of Dahl SS rats, which are thought to contribute to the development of hypertension and renal disease.