Division of Nephrology, Beth Israel Medical Center, 330 Brookline Avenue, Boston, MA 02115, USA.
J Am Soc Nephrol. 2012 Nov;23(11):1879-90. doi: 10.1681/ASN.2012030323. Epub 2012 Sep 20.
Rare loss-of-function mutations in the calcium-sensing receptor (Casr) gene lead to decreased urinary calcium excretion in the context of parathyroid hormone (PTH)-dependent hypercalcemia, but the role of Casr in the kidney is unknown. Using animals expressing Cre recombinase driven by the Six2 promoter, we generated mice that appeared grossly normal but had undetectable levels of Casr mRNA and protein in the kidney. Baseline serum calcium, phosphorus, magnesium, and PTH levels were similar to control mice. When challenged with dietary calcium supplementation, however, these mice had significantly lower urinary calcium excretion than controls (urinary calcium to creatinine, 0.31±0.03 versus 0.63±0.14; P=0.001). Western blot analysis on whole-kidney lysates suggested an approximately four-fold increase in activated Na(+)-K(+)-2Cl(-) cotransporter (NKCC2). In addition, experimental animals exhibited significant downregulation of Claudin14, a negative regulator of paracellular cation permeability in the thick ascending limb, and small but significant upregulation of Claudin16, a positive regulator of paracellular cation permeability. Taken together, these data suggest that renal Casr regulates calcium reabsorption in the thick ascending limb, independent of any change in PTH, by increasing the lumen-positive driving force for paracellular Ca(2+) transport.
钙敏感受体 (Casr) 基因的罕见失功能突变导致甲状旁腺激素 (PTH) 依赖性高钙血症时尿钙排泄减少,但 Casr 在肾脏中的作用尚不清楚。使用受 Six2 启动子驱动的 Cre 重组酶表达的动物,我们生成了 Casr 在肾脏中几乎检测不到 mRNA 和蛋白的小鼠。这些小鼠的基础血清钙、磷、镁和 PTH 水平与对照小鼠相似。然而,当用膳食钙补充剂挑战时,与对照组相比,这些小鼠的尿钙排泄明显减少(尿钙与肌酐的比值为 0.31±0.03 对 0.63±0.14;P=0.001)。对全肾裂解物进行的 Western blot 分析表明激活的 Na(+)-K(+)-2Cl(-)共转运蛋白 (NKCC2) 增加了约四倍。此外,实验动物还表现出 Claudin14 的显著下调,Claudin14 是厚升支中细胞旁阳离子通透性的负调节剂,以及 Claudin16 的微小但显著上调,Claudin16 是细胞旁阳离子通透性的正调节剂。这些数据表明,肾脏 Casr 通过增加细胞旁 Ca(2+)转运的管腔正驱动力,独立于 PTH 的任何变化,调节厚升支中的钙重吸收。