PRDM16 orchestrates neuron-vascular communication for angiogenesis during brain development

Purpose:To gain a deeper insight into how PRDM16 regulates neuron-vascular communication for angiogenesis, RNA-sequencing (RNA-seq) was performed to analyze the genome-wide changes by PRDM16 deletion at E15. Methods: Total RNA was extracted from E15 telencephalic tissue of Prdm16cKO and Prdm16fl/fl mice. Then 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 Illumina HiSeq 2500 platform in Annoroad Genomics. Results: Approximately approximately one thousand transcripts showed differential expression between the Prdm16fl/fl and Prdm16cKO brain cortex, with a fold change ≥1.5 and p value <0.05. Gene ontology (GO) analysis showed that the down-regulated genes were enriched in the terms related to neurogenesis, blood vessel development and secretion by cells. Up-regulated genes showed a significant enrichment of terms involved in negative regulation of cell communication and negative regulation of angiogenesis. These results reflected the importance of PRDM16 in cortical development. Conclusions: We conclude that RNA-seq based transcriptome characterization would provide a framework for understanding how Prdm16 gene contribute to brain cortical development. Total RNA was extracted from E15 telencephalic tissue of Prdm16cKO and Prdm16fl/fl mice and generated by deep sequencing, in triplicate, using Illumina HiSeq 2500.

研究目的：为深入解析PRDM16如何调控神经元-血管通讯以促进血管生成，本研究通过RNA测序（RNA-seq）分析了胚胎第15天（E15）时点PRDM16敲除后全基因组的表达变化。实验方法：从Prdm16cKO与Prdm16fl/fl小鼠的E15端脑组织中提取总RNA，采用安捷伦2100生物分析仪对总RNA进行质量检测与定量。将总RNA反转录为cDNA并构建测序文库后，于诺禾致源（Annoroad Genomics）使用Illumina HiSeq 2500平台完成高通量测序。实验结果：在Prdm16fl/fl与Prdm16cKO小鼠大脑皮层中，约有1000个转录本呈现出差异表达（倍数变化≥1.5，p值<0.05）。基因本体（Gene Ontology, GO）富集分析显示，下调基因显著富集于神经发生、血管发育及细胞分泌相关的功能条目；而上调基因则显著富集于细胞通讯负调控与血管生成负调控相关的功能条目。上述结果揭示了PRDM16在大脑皮层发育过程中的重要作用。研究结论：本研究通过基于RNA测序的转录组表征，为解析PRDM16基因如何参与大脑皮层发育提供了研究框架。本次测序以Prdm16cKO与Prdm16fl/fl小鼠的E15端脑组织为材料，采用Illumina HiSeq 2500平台完成了三次生物学重复的深度测序。