Transcription profiling of mouse Clcn5 knockout in proximal tubules of the kidney

Dent disease has multiple defects attributed to proximal tubule malfunction including low molecular weight proteinuria, aminoaciduria, phosphaturia and glycosuria. In order to understand the changes in kidney function of the Clc5 transporter gene knockout mouse model of Dent disease, we examined gene expression profiles from proximal tubules of mouse kidneys. Overall 720 genes are expressed differentially in the proximal tubules of the Dent Clcn5 knockout mouse model compared to those of control wild type mice. The fingerprint of these gene changes may help us to understand the phenotype of Dent disease. Experiment Overall Design: Renal proximal tubules were dissected from wild type and Clcn5 knockout mice. Mice were anesthetized with halothane, the abdominal aorta of each animal was accessed and the left kidney was perfused with an ice-cold salt. Proximal tubule dissection was performed in an ice-cold salt solution. After dissection of approximately 80-100 segments of 2 mm in length per kidney, the RNA for 3-4 mice was combined to have enough RNA per chip. Experiment Overall Design: 3 microarrays each of wild type and knockout mouse proximal tubule were processed丹特病（Dent disease）是一类因近端肾小管功能异常导致的多缺陷综合征，临床表现包括低分子量蛋白尿（low molecular weight proteinuria）、氨基酸尿（aminoaciduria）、磷酸盐尿（phosphaturia）及糖尿（glycosuria）。为探究丹特病Clc5转运蛋白（Clc5 transporter）基因敲除小鼠模型的肾功能变化，我们对该模型小鼠肾脏近端肾小管的基因表达谱进行了检测。相较于野生型（wild type）对照小鼠，丹特病Clcn5基因敲除小鼠的近端肾小管中共有720个基因呈现差异表达。上述基因表达变化特征或可为解析丹特病的表型提供重要参考。 实验整体设计： 从野生型与Clcn5基因敲除小鼠体内分离肾近端肾小管。小鼠经氟烷（halothane）麻醉后，暴露腹主动脉，以冰冷盐溶液灌注左侧肾脏。随后在冰冷盐溶液中进行近端肾小管分离操作：每只肾脏分离得到约80~100段长度为2mm的肾小管片段后，将3~4只小鼠的样本RNA合并，以满足单张芯片检测所需的RNA量。 实验整体设计： 分别对野生型与基因敲除小鼠的近端肾小管样本各制备3张微阵列（microarrays）芯片并完成检测流程。