Endocardial Notch Signaling Promotes Cardiomyocyte Proliferation in the Regenerating Zebrafish Heart through Wnt Pathway Antagonism

The goal of this study is to understand the molecular mechanisms underlying this Notch-induced muscle cell proliferation defect, we sequenced the transcriptome of control ventricles and Notch-suppressed ventricle at 5 days post-injury, when cardiomyocyte proliferation is normally robust. In order to identify downstream targets of Notch signaling of cardimyotyes proliferation during zebrafish heart regeneration, we compared transcriptome profiles between wildtype hearts and DN-MAML hearts by RNA sequencing strategy. The heart samples were collected, RNA extracted, and cDNA generated for library construction. The samples were collected in triplicate and sequenced at the PCPGM on an Illumina HiSeq 2000.

本研究旨在阐明Notch介导的心肌细胞增殖缺陷背后的分子机制。我们于损伤后5天——该时间点心肌细胞增殖本应处于旺盛水平——对对照组心室与Notch抑制型心室的转录组（transcriptome）进行了测序。为鉴定斑马鱼心脏再生过程中调控心肌细胞增殖的Notch信号通路下游靶标，我们通过RNA测序（RNA sequencing）技术比较了野生型心脏与DN-MAML心脏的转录组特征。实验中我们收集心脏样本、提取总RNA并反转录生成cDNA以构建测序文库，所有样本均设置三次生物学重复，并于PCPGM实验室使用Illumina HiSeq 2000测序平台完成测序。