端脑发育中骨发生形态蛋白对转录因子ISL-1调节作用的研究

发布时间：2018-04-12 20:29

本文选题：端脑 + 胆碱能神经元　；参考：《吉林大学》2006年硕士论文

【摘要】：胆碱能神经元参与机体学习、记忆和运动等功能。广泛分布于中枢神经系统脊髓、脑干、大脑皮质及基底神经核内。生物体内的多种行为功能紊乱均涉及到胆碱能系统病变。因此,认识和了解胆碱能神经元的发生发育规律是攻克这些相关疾病的基础。近年来,对端脑发育的研究无论是细胞水平还是分子水平均有很大进展,但是对于胆碱能神经元发生发育机制的研究知之甚少。前期工作证实在神经元发生期,端脑脑室区放射状胶质细胞(Radial glia)是胆碱能神经元的祖细胞。胆碱能神经元的标志性蛋白是乙酰胆碱转移酶(ChAT),在小鼠胚胎12天ChAT开始表达。2000年IgnacioLópez-Coviella等在SCIENCE杂志中报道了低浓度BMP9在体内和体外均可直接作用于E14小鼠端脑ChAT反应细胞,上调乙酰胆碱转移酶的表达,提示BMPs可以诱导胆碱能神经元的发生。我们认为BMPs作为细胞外信息分子与细胞内同源域转录因子基因共同调控端脑神经元的分化,决定神经前体细胞命运。根据资料分析,控制胆碱能神经元发生的主要同源域转录因子可能是ISL-1,因为ISL-1表达于发育中的胆碱能神经元内,维持发育中胆碱能神经元表型。此外ISL-1与特定DNA序列的结合位点存在于ChAT基因座启动子区内。因此,ISL1可能是与BMPs诱导胆碱能表型相关的同源域转录因子, BMPs可能于ISL-1共同作用进而诱导放射状胶质细胞向胆碱能神经元分化。为了证明此假设,本实验通过对E12、E14、E18大鼠胚胎端脑细胞进行体外培养观察,改变培养液中BMP4浓度,并采用RT-PCR技术,以确定BMP4对端脑细胞的影响及胆碱能前体细胞ISL-1 mRNA表达是否随之发生变化。细胞培养结果显示,BMP4对E14、E18大鼠端脑体外培养细胞均有促进其聚集成团以及促使细胞团之间伸出突起的作用。RT-PCR检测显示: 20ng/ml BMP4使细胞中ISL-1 mRNA表达量有明显增加, 40ng/ml BMP4使细胞中ISL-1 mRNA表达量减少,提示ISL-1可能是与BMPs共同诱导胆碱能表型的主要同源域转录因子。
[Abstract]:Cholinergic neurons are involved in learning, memory and motor functions.Widely distributed in the central nervous system spinal cord, brain stem, cerebral cortex and basal nucleus.A variety of behavioral disorders in organisms are associated with cholinergic system lesions.Therefore, understanding and understanding the occurrence and development of cholinergic neurons is the basis of these diseases.In recent years, much progress has been made in the study of telencephalon development, both at cellular and molecular levels, but little is known about the mechanism of cholinergic neuron development.It was confirmed that radiative glia was the progenitor of cholinergic neurons during neuronogenesis.The iconic protein of cholinergic neurons is acetylcholine transferase (acetylcholine transferase) ChATA, which is expressed at the 12th day of mouse embryo. In 2000, IgnacioL 贸 pez-Coviella et al reported in the SCIENCE journal that low concentration of BMP9 could directly act on ChAT response cells of the terminal brain of E14 mice, both in vivo and in vitro.Upregulation of acetylcholine transferase expression suggests that BMPs can induce cholinergic neurons.We believe that BMPs acts as an extracellular pheromone and an intracellular homologous domain transcription factor gene to regulate the telencephalon.The differentiation of neurons determines the fate of neural precursor cells.According to the data analysis, the main homologous transcription factor of cholinergic neurons is probably ISL-1, because ISL-1 is expressed in developing cholinergic neurons and maintains the phenotype of developing cholinergic neurons.In addition, the binding sites of ISL-1 to specific DNA sequences are located in the promoter region of ChAT locus.Therefore, ISL1 may be a homologous domain transcription factor associated with BMPs induced cholinergic phenotype, and BMPs may interact with ISL-1 to induce radiative glial cells to differentiate into cholinergic neurons.In order to prove this hypothesis, the BMP4 concentration in the culture medium was changed by in vitro culture of E12E14E18 rat embryonic brain cells, and RT-PCR technique was used.To determine the effect of BMP4 on the terminal brain cells and whether the expression of ISL-1 mRNA in cholinergic precursor cells changes with it.The results of cell culture showed that BMP4 could promote the aggregation and protruding of endencephalon of E14 / E18 rat in vitro. RT-PCR showed that 20ng/ml BMP4 increased the expression of ISL-1 mRNA and 40ng/ml.BMP4 reduced the expression of ISL-1 mRNA in cells.These results suggest that ISL-1 may be a major homologous domain transcription factor co-inducing cholinergic phenotype with BMPs.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329

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