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Zinc‐finger nuclease knockout of dual‐specificity protein phosphatase‐5 protects against DOCA‐salt induced chronic renal injury
Author(s) -
Fan Fan,
Murphy Sydney R,
Ge Ying,
Mims Paige N,
Maeda Kenji J,
Williams Jan M,
Roman Richard J
Publication year - 2017
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.31.1_supplement.1015.11
Subject(s) - medicine , knockout mouse , endocrinology , chemistry , kidney , nuclease , receptor , gene , biochemistry
We recently reported that zinc‐finger nuclease knockout (KO) of dual‐specificity protein phosphatase‐5 (Dusp5) enhances the myogenic response and autoregulation of cerebral blood flow in FHH.1 BN rats. We identified 4 single nucleotide polymorphisms in Dusp5 in FHH as compared with BN rats, two of which alter CpG sites and another one causes G155R mutation. In the present study, we evaluated whether Dusp5 also plays a role in the regulation of autoregulation of renal blood flow and renal function in a Dusp5 knockout (KO) rat in the FHH.1 BN genetic background, which an 11 bp frame‐shift deletion is introduced by zinc‐finger nuclease and a premature stop codon at AA 121 of Dusp5 protein is produced. The expression of Dusp5 was lower in FHH and in Dusp5 KO rats in comparison to the FHH.1 BN rats. The level of phosphorylated ERK1/2 (p‐ERK1/2), a substrate of Dusp5, was enhanced in the KO rats. The blood pressure and urinary protein excretion was similar in Dusp5 KO (n = 43) and FHH.1 BN rats (n = 30) when they were at 12‐week of age (114 ± 2 vs. 115 ± 1 mmHg and 32 ± 2 vs. 29 ± 3 mg/day). In response to an elevation in pressure from 60 to 120 mm Hg, the renal afferent arterioles (Af‐arts) constricted by 29 ± 2% and 11 ± 1%, and the vascular tone was increased to 36 ± 2% vs 19 ± 1%, respectively in Dusp5 KO (n = 6) in comparison to FHH.1 BN rats (n = 17). Blood pressure was 169 ± 5 vs. 177 ± 5 mmHg in Dusp5 KO (n = 11) vs. FHH.1 BN rats (n = 14) after 3‐week DOCA‐salt treatment. However, proteinuria was significantly reduced in Dusp5 KO rats (258 ± 22 mg/day, n = 12) in comparison to FHH.1 BN rats (338 ± 30 mg/day, n = 12) with the same treatment. The renal injury induced by DOCA‐salt was markedly reduced in Dusp5 KO vs. FHH.1 BN rats as the glomerular injury scores were 2.49 ± 0.03 vs. 2.54 ± 0.01; the areas of renal fibrosis were 5.5 ± 0.5% vs. 8.3 ± 0.3%; the areas of renal protein casts were 0.5 ± 0.2% vs. 2.4 ± 1.0% and renal arteriolar media‐to‐lumen ratios were 0.64 ± 0.03 vs. 0.81 ± 0.04, respectively. These results indicate that Dusp5 modulates myogenic reactivity in the renal circulation and protects against DOCA‐salt hypertension induced chronic renal injury. It supports the view that upregulation of the expression of Dusp5 and/or an activating mutation in Dusp5 may contribute to the impaired myogenic response and autoregulation of renal blood flow and promote the development of chronic renal injury in FHH rats. Support or Funding Information This study was supported by grants HL36279 (RJR) and DK104184 (RJR), AG050049 (FF), P20GM104357 (RJR, FF and JMW) from the National Institutes of Health; 16GRNT31200036 (FF) and 12SDG9440034 (JMW) from the American Heart Association. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.