在端脑发育中骨形态发生蛋白-4对同源域转录因子Nkx2.1表达的影响

发布时间：2017-12-28 16:24

本文关键词：在端脑发育中骨形态发生蛋白-4对同源域转录因子Nkx2.1表达的影响　出处：《吉林大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的:研究在端脑发育中骨形态发生蛋白4(BMP-4)对同源域转录因子Nkx2.1表达的影响,探讨Nkx2.1在胆碱能神经元发生过程中的作用以及能否被BMP-4所诱导。方法:将E16大鼠胚胎端脑细胞做体外无血清原代培养,用不同浓度的BMP-4诱导胆碱能神经元的分化,采取免疫细胞化学染色实验观察不同浓度BMP-4对ChAT阳性免疫反应细胞数量的影响。在实验过程中采取对胆碱能神经元分化影响最大的BMP-4浓度,加入到体外培养的E16大鼠胚胎端脑细胞中,进行RT-PCR检测,以研究Nkx2.1在端脑发育中BMP-4对Nkx2.1基因表达的影响。结果:不同浓度BMP-4对胆碱能神经元发生的影响有所不同。低浓度(10ng/ml、20ng/ml)BMP-4可使ChAT阳性细胞数量增多;高浓度(40ng/ml)BMP-4可使ChAT阳性细胞数量减少。RT-PCR结果显示实验组和对照组Nkx2.1基因表达没有明显区别。结论:(1)在体外培养的E16端脑细胞中,低浓度(20ng/ml)BMP-4能促进胆碱能前体细胞的发生和分化;(2)在体外培养的E16端脑细胞中,高浓度(40ng/ml)BMP-4能抑制胆碱能前体细胞的发生和分化;(3)BMP-4细胞外信号分子不能调节E16端脑细胞Nkx2.1 mRNA的表达。
[Abstract]:Objective: To study the bone morphogenetic protein 4 in telencephalon development (BMP-4) on expression of homeodomain transcription factor Nkx2.1, to investigate the effects of Nkx2.1 on cholinergic neurons during the process of action and can be induced by BMP-4. Methods: embryonic stem cells from E16 rats were cultured in vitro without serum. Primary cultures of cholinergic neurons were induced by different concentrations of BMP-4. Immunocytochemical staining was used to observe the effect of different concentrations of BMP-4 on the number of ChAT positive immunoreactive cells. During the experiment, we took the most influential BMP-4 concentration on the cholinergic neurons, and added it to the embryonic brain cells of E16 rats cultured in vitro. RT-PCR was used to detect the effect of BMP-4 on Nkx2.1 gene expression in the development of Nkx2.1. Results: the effects of different concentrations of BMP-4 on the occurrence of cholinergic neurons were different. Low concentration (10ng/ml, 20ng/ml) BMP-4 can increase the number of ChAT positive cells, and high concentration (40ng/ml) BMP-4 can reduce the number of ChAT positive cells. The results of RT-PCR showed that there was no significant difference in the expression of Nkx2.1 gene between the experimental group and the control group. Conclusion: (1) in vitro of E16 brain cells, low concentration (20ng/ml) of BMP-4 can promote the occurrence and differentiation of cholinergic progenitor cells; (2) in vitro of E16 brain cells, high concentrations of BMP-4 (40ng/ml) can inhibit the occurrence and differentiation of cholinergic progenitor cells; (3) the expression of BMP-4 extracellular signal molecules cannot regulate E16 Nkx2.1 brain cell end mRNA.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R341

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