Gene expression profiling of Wt1 mutant renal anlagen

Wt1 is required for renal development and homozyogus knockout mice show renal agenesis caused by apoptosis of the metanepric mesnchyme. To identify genes regulated by WT1, we performed comparative gene expression profiling on kidney rudiments of Wt1 heterozygous and homozygous mutant E10.25 embryos. To identify deregulated genes, timed matings were performed and embryos isolated at E10.25, a time point before the onset of apoptosis in Wt1 mutants. The region containing the metanephric mesenchyme was microdissected, RNA isolated and after labeling, hybridized to Mouse Genome Survey Microarrays. Deregulated genes were identified using GeneSpring (GSX) software.Wt1 是肾脏发育所必需的基因，纯合敲除小鼠会出现因后肾间充质（metanephric mesenchyme）凋亡导致的肾发育不全。为鉴定受Wt1调控的基因，本研究针对Wt1杂合及纯合突变的E10.25胚胎的肾原基开展了比较基因表达谱分析。为筛选表达失调的基因，研究人员通过定时交配获得胚胎，并在Wt1突变体凋亡发生前的时间节点E10.25处分离胚胎。随后对包含后肾间充质的区域进行显微切割，提取RNA并完成标记后，将其与小鼠基因组探查微阵列（Mouse Genome Survey Microarrays）进行杂交。最终利用GeneSpring (GSX) 软件鉴定出了表达失调的基因。