Cx43基因敲除鼠胚心脏近端流出道组织中Galpha13信号通路基因的差异表达

Differentially Expressed Genes Related with Galpha13 Pathway in the Cardiac Outflow Tract Tissue of Cx43 Knockout Mice

目的研究Cx43基因剔除(Cx43KO)小鼠胚胎心脏近端流出道组织中基因表达谱的改变,筛选可能导致Cx43KO小鼠流出道梗阻的相关基因。方法以胎龄(embryonic day,ED)14.5天的Cx43KO和野生型(Cx43WT)鼠胚心脏近端流出道部分为研究对象,分别提取总RNA,逆转录成cDNA;并在体外转录为cRNA,同时进行生物素标记及片段化;再与Affymetrix-4302.0基因芯片进行杂交。杂交信号经扫描后,应用相关生物信息软件分析基因表达情况。结果与Cx43WT组相比,Cx43KO组中表达上调2倍以上的基因共有287个,表达下调2倍以上的基因有199个。其中表达差异的基因参与转录调控、细胞周期等主要生理过程。进一步筛查表达差异1.5倍以上的基因发现,Galpha13信号通路上的多个基因在Cx43KO组有明显变化。结论利用基因芯片技术初步筛选出与Cx43KO鼠胚心脏近端流出道发育有关的多个基因,其中Galpha13信号通路上的相关基因可能与Cx43KO小鼠流出道梗阻的发生有关。

Objective To investigate the changes of gene expression in the cardiac outflow tract （OFF） in Cx43 knockout mouse embryo, and to elucidate the genes involved in the pathogenesis of pulmonary outflow obstruction. Methods The cDNA was retrotranscripted from RNA extracted from OFF tissues of both Cx43 knockout and Cx43 wild type mouse embryos on embryonic day （ED） 14.5. The biotin-labeled cRNA derived from the transcription of cDNA was fragmented as probes. The probes were hybridized with Affymetrix Mouse Genome 430 2.0 Array. Gene Array Scanner was used to screen the signals of hybridization and the expression of genes was detected. Results Among the differentially expressed genes, there were 287 upregulated and 199 downregulated in Cx43 knockout OFF tissue as compared with those in Cx43 wild type heart. Functions of proteins encoded by the altered genes encompassed all functional categories, with largest percentage in genes involved in regulation of transcription, cell cycle, etc. Among the differentially expressed genes in the Cx43 knockout heart were those related to Galphal3 signaling pathway. Conclusions Genes related with Galphal3 signaling pathway differently expressed in ED14.5 Cx43 knockout OFF tissue may be involved in the pathogenesis of pulmonary outflow obstruction.