Pd1 regulates proliferation and differentiation of neural progenitors during brain development

To gain a total insight into how Wt1 regulates embryonic neurogenesis, RNA-seq was performed to compare the different expressed genes at E13. Approximately approximately one thousand transcripts showed differential expression between the Pd1fl/fl and Pd1CKO mice brain cortex, with a fold change >=1.5 and p value <0.05. These results indicated the importance of Pd1 in cortical development. RNA-seq based transcriptome characterization would provide a global understanding how Pd1 gene contribute to brain cortical development. Total RNA were extracted from E13 telencephalic tissue of 2 Pd1fl/fl and 2 Pd1CKO mice. Then the total RNA was quality controlled and quantified using an Agilent 2100 Bioanalyzer. After converting to cDNA and building library, high-throughput sequencing was performed using the Ptc 225 Peltier Thermal Cycler.为全面解析Wt1调控胚胎神经发生的具体机制，本研究采用RNA测序（RNA-seq）技术，比较胚胎期13天（E13）样本间的差异表达基因。在Pd1fl/fl与Pd1CKO小鼠的大脑皮层中，约有1000个转录本呈现显著表达差异，其表达折叠倍数≥1.5且p值<0.05。上述结果证实了Pd1在大脑皮层发育过程中的关键作用。基于RNA-seq的转录组特征分析，可为解析Pd1基因参与大脑皮层发育的分子机制提供全局视角。本研究从2只Pd1fl/fl与2只Pd1CKO小鼠的E13端脑组织中提取总RNA，随后采用安捷伦2100生物分析仪（Agilent 2100 Bioanalyzer）对总RNA进行质量质控与定量检测。经反转录合成cDNA并构建测序文库后，使用Ptc 225珀尔帖热循环仪（Ptc 225 Peltier Thermal Cycler）完成高通量测序。