Cowley Allen W, Yang Chun, Kumar Vikash, Lazar Jozef, Jacob Howard, Geurts Aron M, Liu Pengyuan, Dayton Alex, Kurth Theresa, Liang Mingyu
Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin;
Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin;
Physiol Genomics. 2016 Jan;48(1):62-72. doi: 10.1152/physiolgenomics.00097.2015. Epub 2015 Nov 3.
A 1.37 Mbp region of chromosome 13 previously identified by exclusion mapping was consistently associated with a reduction of salt-induced hypertension in the Dahl salt-sensitive (SS) rat. This region contained five genes that were introgressed from the salt-insensitive Brown Norway (BN) rat. The goal of the present study was to further narrow that region to identify the gene(s) most likely to protect from salt-induced hypertension. The studies yielded a subcongenic SS rat strain containing a 0.71 Mbp insert from BN (26-P strain) in which salt-induced hypertension was reduced by 24 mmHg. The region contained two protein-coding genes (Astn1 and Pappa2) and a microRNA (miR-488). Pappa2 mRNA in the renal cortex of the protected 26-P was 6- to 10-fold greater than in SS fed a 0.4% NaCl diet but was reduced to levels observed in SS when fed 8.0% NaCl diet for 7 days. Compared with brain nuclei (NTS, RVLM, CVLM) and the adrenal gland, Pappa2 in the renal cortex was the only gene found to be differentially expressed between SS and 26-P and that responded to changes of salt diet. Immunohistochemistry studies found Pappa2 localized in the cytosol of the epithelial cells of the cortical thick ascending limbs. In more distal segments of the renal tubules, it was observed within tubular lumens and most notably bound to the apical membranes of the intercalated cells of collecting ducts. We conclude that we have identified a variant form of Pappa2 that can protect against salt-induced hypertension in the Dahl S rat.
先前通过排除定位法确定的13号染色体上一个1.37兆碱基对的区域,始终与降低Dahl盐敏感(SS)大鼠的盐诱导高血压有关。该区域包含五个从盐不敏感的棕色挪威(BN)大鼠导入的基因。本研究的目的是进一步缩小该区域,以确定最有可能预防盐诱导高血压的基因。这些研究产生了一种亚同源SS大鼠品系,其含有来自BN的0.71兆碱基对插入片段(26-P品系),其中盐诱导的高血压降低了24毫米汞柱。该区域包含两个蛋白质编码基因(Astn1和Pappa2)和一个微小RNA(miR-488)。受保护的26-P品系肾皮质中的Pappa2 mRNA比喂食0.4% NaCl饮食的SS大鼠高6至10倍,但在喂食8.0% NaCl饮食7天后降至SS大鼠中观察到的水平。与脑核(NTS、RVLM、CVLM)和肾上腺相比,肾皮质中的Pappa2是唯一在SS和26-P之间差异表达且对盐饮食变化有反应的基因。免疫组织化学研究发现Pappa2定位于皮质厚升支上皮细胞的细胞质中。在肾小管的更远端部分,在管腔内观察到它,最显著的是它与集合管闰细胞的顶端膜结合。我们得出结论,我们已经鉴定出一种Pappa2的变体形式,它可以预防Dahl S大鼠的盐诱导高血压。