Experimental Nephrology, Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Fetscherstr. 74, 01307, Dresden, Germany.
Pflugers Arch. 2017 Oct;469(10):1349-1357. doi: 10.1007/s00424-017-1994-y. Epub 2017 May 22.
We reported earlier that PPAR-gamma regulates renin transcription through a human-specific atypical binding sequence termed hRen-Pal3. Here we developed a mouse model to investigate the functional relevance of the hRen-Pal3 sequence in vivo since it might be responsible for the increased renin production in obesity and thus for the development of accompanying arterial hypertension. We used bacterial artificial chromosome construct and co-placement strategy to generate two transgenic mouse lines expressing the human renin gene from identical genomic locus without affecting the intrinsic mouse renin expression. One line carried a wild-type hRen-Pal3 in the transgene (Pal3wt strain) and the other a mutated non-functional Pal3 (Pal3mut strain). Human renin expression was correctly targeted to the renin-producing juxtaglomerular (JG) cells of kidney in both lines. However, Pal3mut mice had lower basal human renin expression. Since human renin does not recognize mouse angiotensinogen as substrate, the blood pressure was not different between the strains. Stimulation of renin production with the angiotensin-converting enzyme inhibitor enalapril equipotentially stimulated the human renin expression in Pal3wt and Pal3mut mice. High-fat diet for 10 weeks which is known to activate PPAR-gamma failed to increase human renin mRNA in kidneys of either strain. These findings showed that the human renin PPAR-gamma-binding sequence hRen-Pal3 is essential for basal renin expression but dispensable for the cell-specific and high-fat diet regulated renin expression in the kidney.
我们之前报道过,过氧化物酶体增殖物激活受体-γ(PPAR-γ)通过一种称为 hRen-Pal3 的人类特异性非典型结合序列来调节肾素转录。在这里,我们开发了一种小鼠模型来研究 hRen-Pal3 序列在体内的功能相关性,因为它可能是肥胖症中肾素产生增加的原因,从而导致伴随的动脉高血压的发生。我们使用细菌人工染色体构建体和共定位策略,在不影响内在的小鼠肾素表达的情况下,从相同的基因组位点产生表达人类肾素基因的两个转基因小鼠品系。一条线携带野生型 hRen-Pal3 转基因(Pal3wt 品系),另一条携带突变的无功能 Pal3(Pal3mut 品系)。人类肾素在两条线的肾素产生的肾小球旁器(JG)细胞中被正确靶向表达。然而,Pal3mut 小鼠的基础人类肾素表达较低。由于人类肾素不识别小鼠血管紧张素原作为底物,因此两种品系之间的血压没有差异。血管紧张素转换酶抑制剂依那普利的刺激等效地刺激了 Pal3wt 和 Pal3mut 小鼠的人类肾素表达。已知激活过氧化物酶体增殖物激活受体-γ的高脂肪饮食在 10 周内未能增加两种品系肾脏中的人类肾素 mRNA。这些发现表明,人类肾素 PPAR-γ 结合序列 hRen-Pal3 对于基础肾素表达是必需的,但对于肾脏中细胞特异性和高脂肪饮食调节的肾素表达是可有可无的。