Islet-1在心脏发育的乙酰化调控网络中的枢纽作用

发布时间：2018-04-10 03:14

本文选题：Islet-1　切入点：心脏发育　出处：《重庆医科大学》2012年硕士论文

【摘要】：目的：阐明小鼠胚胎心脏发育过程中转录因子Islet-1的时序表达规律，并探讨其在组蛋白乙酰化调控网络中的枢纽作用。方法:选取胎龄11.5天（E11.5）、14.5天（E14.5）、17.5天（E17.5）的胎鼠心脏和新生鼠心脏，利用Western-blot方法定量分析小鼠胚胎心脏发育过程中，转录因子Islet-1的时序性表达规律，并分别利用蛋白质免疫共沉淀（Co-IP），染色质免疫共沉淀（Ch-IP），Q-PCR和聚丙烯酰胺凝胶电泳方法，探讨Islet-1与组蛋白乙酰化调控网络中其他相关蛋白、基因的关系。结果：（1）转录因子Islet-1的表达在小鼠胚胎心脏发育过程中呈先增强后减弱趋势，在E14.5表达最高，与其他时间点相比明显升高（P0.05），但其他时间点（E11.5， E17.5和新生鼠）之间表达差异没有统计学意义。（2）胚胎心脏发育过程中，转录因子Islet-1与组蛋白乙酰化酶p300、GCN5、PCAF和Ac-H3相互结合以蛋白复合物形式存在。（3）转录因子Islet-1结合于心脏发育相关基因GATA4，Mef2c和Tbx5的启动子区域，，可能参与调控基因的表达。结论：在小鼠胚胎心脏发育过程中，转录因子Islet-1可能作为枢纽因子，募集组蛋白乙酰化酶p300、GCN5、PCAF和Ac-H3，介导组蛋白乙酰化修饰对心脏发育相关基因GATA4，Mef2c和Tbx5的调控作用。
[Abstract]:Aim: to elucidate the temporal expression of transcription factor Islet-1 during the development of mouse embryonic heart and to explore its pivotal role in histone acetylation regulatory network.Methods: fetal rat heart and newborn rat heart were selected at 11.5 days of gestational age. The expression of transcription factor Islet-1 was quantitatively analyzed by Western-blot method during the development of mouse embryonic heart.The relationship between Islet-1 and other related proteins and genes in histone acetylation regulatory network was investigated by protein immunoprecipitation Co-IPN, chromatin immunoprecipitation Q-PCR and polyacrylamide gel electrophoresis.Results the expression of Islet-1 increased first and then decreased during the development of mouse embryonic heart, and the highest expression was found in E14.5.Compared with other time points, the expression of E11.5, E17.5 and newborn mice was significantly higher than that at other time points, but there was no significant difference in the expression of E11.5, E17.5 and newborn mice.Transcription factor Islet-1 binds to histone acetylase p300GCN5PCAF and Ac-H3 in the form of protein complex. The transcription factor Islet-1 binds to the promoter region of cardiac development-related gene GATA4Mef2c and Tbx5, which may be involved in the regulation of gene expression.Conclusion: during the development of mouse embryonic heart, transcription factor Islet-1 may act as a pivotal factor, and histone acetylase p300GCN5PCAF and Ac-H3 may mediate the regulation of cardiac development-related genes GATA4Mef2c and Tbx5 by histone acetylation modification.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R321

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