室间隔缺损ALK3下游相关基因的初步研究

Preliminary Study of ALK3 Downstream Genes Related to Ventricular Septum Defect

了解ALK3在心脏发育中的作用 ,探索室间隔缺损的特异相关基因及其信号传导途径。应用α MHC Cre loxP系统 ,建立了心脏ALK3基因敲除小鼠模型 ,利用PCR选择性cDNA差异显示法和基因芯片扫描 (RNAmicroar ray)的方法 ,比较对照组和试验组mRNA表达水平 ,筛选ALK3下游基因。对照组的mRNA来自α MHC Cre+ -ALK3F + 的 11.5d小鼠胚胎心脏 ,试验组的mRNA来自α MHCCre+ - ALK3F - 的 11.5d小鼠胚胎心脏。心脏特异的ALK3基因敲除后 ,血小板激活因子乙酰水解酶及转录因子Pax 8等基因的表达水平下降 ,β亚类 14 3 3蛋白及蛋白酪氨酸激酶等基因的表达水平上调。血小板激活因子乙酰水解酶及转录因子Pax 8等基因可能是ALK3重要的下游基因 ,与室间隔缺损的形成有关 ;β亚类 14 3

To investigate the function of ALK3 gene, the gene regulation and the signaling pathway related to ventricular septum defect during heart development. The model mice with ALK3 gene knock out via α MHC Cre/lox P system were bred. The mRNA expression level of control group was compared with that of experiment group and ALK3 downstream genes were screened using PCR select cDNA subtraction microarray. The mRNA of control group was extracted from E11.5 normal mouse hearts, and that of experiment group, from E11.5 hearts of mice with α MHC Cre +/- ALK3 F/+ genotype. It was found that the mice with ALK3 gene knock out produced heart defects involving the interventricular septum. The platelet activating factors acetylhydrolase and the transcription factor Pax 8 and so on, were down regulated. However, the Protein Tyrosine Kinase (PTK) of Focal Adhesion Kinase (FAK) subfamily and beta subtype protein 14 3 3 were up regulated in the α MHC Cre +/- ALK3 F/- mice. These data provide support that ALK3 gene played an important role during heart development. The platelet activating factors acetylhydrolase and Pax 8 genes could be important ALK3 downstream genes in the BMP signaling pathway during interventricular septum development. PTK and beta subtype protein 14 3 3 might be regulatory factors in this pathway.