辅助生殖技术对小鼠脑组织及人胚胎中GABRB3基因表达的影响

项目名称： 辅助生殖技术对小鼠脑组织及人胚胎中GABRB3基因表达的影响

项目编号： No.81501323

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 付磊

作者单位： 首都医科大学

项目金额： 17万元

中文摘要： 辅助生殖技术的安全性问题受到人们的广泛关注。有报道指出，辅助生殖技术是自闭症的一个风险因素，但也有报道反对这一说法。自闭症是由多基因和环境共同决定的，其中一个候选基因是GABRB3，它编码γ –氨基丁酸（GABA）A受体的β3亚基。制备体内受精、体外受精（IVF）和单精子注射（ICSI）小鼠胚胎，通过自然妊娠或胚胎移植得到后代。利用RT-PCR比较三种小鼠大脑组织的GABRB3基因及相关基因MEPC2、UBE3A的转录水平，根据三种基因转录水平的直观变化推断辅助生殖技术可能对GABRB3基因产生的影响。利用重亚硫酸盐测序和ChIP分析，检测三种样本GABRB3基因启动子区的DNA甲基化水平和组蛋白修饰状态。另一方面，检测发育至不同时期的IVF和ICSI人类胚胎中GABRB3基因的表达情况及其所定位的染色体15q12的倍性，得到GABRB3基因表达的时间图谱。

中文关键词： 辅助生殖技术；自闭症谱系障碍；GABRB3；；小鼠；人

英文摘要： The security of assisted reproductive technology (ART) is attracting more and more attention nowadays. Some reports suggested ART as a risk factor of autism spectrum disorders (ASD), but there are also reports against it. Autism is determined by multiple genes and environment. One of the candidate genes is GABRB3, which encodes beta 3 subunit of the GABA A receptor. Prepare in vivo fertilization, in vitro fertilization and intracytoplasmic sperm injection mouse embryos. Fetus were got through natural pregnancy or embryo transfer. Compare the expression levels of GABRB3, MEPC2 and UBE3A genes in brains of three kinds of mice by RT-PCR. The effect of ART on GABRB3 gene could be inferred according to the visible results. Detect DNA methylation and histone modification status in promoter region of GABRB3 gene by bisulfite sequencing and ChIP analysis. Epigenetic changes could be reversed by epigenetic modification enzyme inhibitors. Change the medication time, dosage and duration of inhibitors, and try to investigate a method with the best inhibitory effect and minimum influence on animal. On the other hand, we investigate GABRB3 expression and chromosome 15q12 ploidy in human embryos produced by IVF or ICSI, and draw the map of GABRB3 expression during embryo development.

英文关键词： assisted reproductive technology;autism spectrum ;GABRB3 ;mouse;human