Direct and indirect podocyte injury manifested in a partial podocytectomy mouse model. Direct and indirect podocyte injury manifested in a partial podocytectomy mouse model

Injured podocytes indirectly influence neighboring podocytes. To study the indirect effect, we generated mosaic mice, Pod-hCD25T/G, in which podocytes express either hCD25 + tdTomato or EGFP alone. A hCD25-targetting immunotoxin LMB2 was injected into the mosaic mice to induce injury selectively in tdTomato(+) podocytes. We analyzed RNA expressing profile in tdTomato(+) (hCD25 expressing) and EGFP(+) (hCD25 not expressing) podocytes before and after LMB2 injection. Overall design: Glomeruli were harvested from Pod-hCD25T/G mouse before and 1, 4, and 7 days after LBM2 treatment. tdTomato(+) and EGFP(+) podocytes were collected by cell sorter and RNA expressing profile was analyzed.

受损足细胞（podocytes）可间接影响邻近的足细胞。为探究该间接作用，我们构建了嵌合小鼠Pod-hCD25T/G，该小鼠的足细胞仅表达人CD25（hCD25）与tdTomato，或仅表达增强绿色荧光蛋白（EGFP）。我们向该嵌合小鼠注射靶向hCD25的免疫毒素LMB2，以选择性诱导tdTomato(+)足细胞发生损伤。我们分别在LMB2注射前后，对tdTomato(+)（表达hCD25）与EGFP(+)（不表达hCD25）足细胞的RNA表达谱进行了分析。整体实验设计：在LBM2处理前及处理后1、4、7天，从Pod-hCD25T/G小鼠体内分离获取肾小球；通过细胞分选仪收集tdTomato(+)与EGFP(+)足细胞，并对其RNA表达谱开展检测分析。