Claudin-10a deficiency shifts proximal tubular Cl- permeability to cation selectivity via claudin-2 translocation

The tight junction (TJ) proteins claudin-2 and -10a form paracellular cation and anion channels, respectively, and have both been reported to be expressed in the proximal tubule (PT). However, the physiological role of claudin-10a in the kidney is yet obscure. Mice deficient in claudin-10a were generated and successful knockout was confirmed by Southern Blot, Western Blot and immunofluorescence staining. The functionality of isolated PTs was investigated electrophysiologically. Compensatory regulation was studied by pharmacological intervention, RNA-Seq analysis, Western Blot and immunofluorescence staining. Overall design: 7 samples from proximal tubule (4 WT, 3 Cldn10a KO), 8 samples from distal convoluted tubule (4 WT, 4 Cldn10a KO)

紧密连接（tight junction, TJ）蛋白闭合蛋白（claudin）-2与-10a分别构成细胞旁阳离子通道与阴离子通道，且二者均已被报道在近端小管（proximal tubule, PT）中表达。然而，闭合蛋白-10a在肾脏中的生理功能仍尚不明确。本研究构建了claudin-10a基因敲除小鼠，并通过Southern印迹（Southern Blot）、蛋白质印迹（Western Blot）及免疫荧光染色证实了基因敲除成功。通过电生理学方法检测了分离得到的近端小管的功能特性，并通过药物干预、RNA测序（RNA-Seq）分析、蛋白质印迹及免疫荧光染色对代偿调控机制进行了研究。实验整体设计：共采集7份近端小管样本（4份野生型（WT）、3份claudin-10a基因敲除型（Cldn10a KO）），以及8份远曲小管（distal convoluted tubule）样本（4份野生型、4份claudin-10a基因敲除型）