本文關(guān)鍵詞： 端腦發(fā)育 膽堿能神經(jīng)元 骨發(fā)生形態(tài)蛋白-4 胰島素基因1增強(qiáng)子蛋白　出處：《吉林大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 由于基底前腦膽堿能神經(jīng)元參與人類運(yùn)動(dòng),學(xué)習(xí)和記憶,又是臨床上多種中樞神經(jīng)系統(tǒng)退行性疾病的靶細(xì)胞,因此,研究誘導(dǎo)膽堿能發(fā)生的關(guān)鍵細(xì)胞外信息分子和控制膽堿能基因座轉(zhuǎn)錄的同源域蛋白對(duì)于揭示膽堿能神經(jīng)元的發(fā)生不僅具有重要的理論意義,而且還有巨大的應(yīng)用價(jià)值。本課題組在前期工作中發(fā)現(xiàn),在E14大鼠端腦細(xì)胞體外培養(yǎng)體系中,20ng／ml BMP4可增加乙酰膽堿轉(zhuǎn)移酶(ChAT)免疫反應(yīng)陽性細(xì)胞數(shù),40ng／ml BMP4可減少ChAT陽性細(xì)胞數(shù)量。此結(jié)果表明不同濃度BMP4對(duì)膽堿能神經(jīng)元發(fā)生和分化具有不同影響。為進(jìn)一步證實(shí)在端腦體外培養(yǎng)體系中不同濃度BMP4對(duì)同源域蛋白ISL-1表達(dá)是否與ChAT的表達(dá)相一致,以推斷膽堿能神經(jīng)元發(fā)生是否通過BMP4信號(hào)傳導(dǎo)途徑作用于同源域蛋白ISL-1而起作用。本實(shí)驗(yàn)采用免疫細(xì)胞化學(xué)和定量分析技術(shù),檢測(cè)了不同濃度BMP4誘導(dǎo)ISL-1蛋白的表達(dá),其結(jié)果顯示,實(shí)驗(yàn)各組均可見同源域蛋白ISL-1在細(xì)胞核中表達(dá)。其中10ng／ml BMP4組ISL-1免疫反應(yīng)陽性細(xì)胞數(shù)與對(duì)照組相比無顯著性差異；20ng／ml BMP4組ISL-1陽性細(xì)胞數(shù)與其它組相比明顯增加,此作用可被BMP抑制劑Noggin阻斷；40ng／ml BMP4組ISL-1陽性細(xì)胞數(shù)顯著減少。此結(jié)果不僅與前期工作中不同濃度BMP4對(duì)ChAT陽性細(xì)胞數(shù)量的影響相一致,而且與不同濃度BMP4誘導(dǎo)ISL-1mRNA表達(dá)的結(jié)果相吻合。說明BMP4信號(hào)傳導(dǎo)途徑不但參與了膽堿能神經(jīng)元發(fā)生過程,同時(shí)可誘導(dǎo)同源域蛋白ISL-1表達(dá)。此結(jié)果還提示BMP4細(xì)胞外信息分子可能通過誘導(dǎo)同源域ISL-1蛋白來調(diào)控端腦神經(jīng)祖細(xì)胞向膽堿能神經(jīng)元分化。
[Abstract]:Because basal forebrain cholinergic neurons are involved in human movement, learning and memory, they are also the target cells for many degenerative diseases of the central nervous system. The study of key extracellular pheromones and homologous proteins that control the transcription of cholinergic loci are not only of theoretical significance in revealing the occurrence of cholinergic neurons, but also in the pathogenesis of cholinergic neurons. Moreover, there is great application value. In our earlier work, we found that, In E14 rat terminal brain cells cultured in vitro, 20ng / ml BMP4 increased the number of acetylcholine transferase (BMP4) immunoreactive cells and 40ng / ml BMP4 decreased the number of ChAT positive cells. In order to further confirm whether the ISL-1 expression of homologous domain protein ISL-1 is consistent with that of ChAT in the culture system of endencephalon in vitro with different concentrations of BMP4. In order to infer whether cholinergic neuronogenesis acts on homologous domain protein ISL-1 through BMP4 signal transduction pathway, the expression of ISL-1 protein induced by different concentrations of BMP4 was detected by immunocytochemistry and quantitative analysis. The results showed that homologous domain protein ISL-1 was expressed in the nucleus of all groups, and the number of ISL-1 immunoreactive cells in 10ng / ml BMP4 group was not significantly different from that in control group. The number of ISL-1 positive cells in 20ng / ml BMP4 group was significantly higher than that in other groups. The number of ISL-1 positive cells in 40 ng / ml BMP4 group blocked by BMP inhibitor Noggin was significantly decreased. The results were not only consistent with the effect of different concentrations of BMP4 on the number of ChAT positive cells in the previous work. These results are consistent with the results of ISL-1mRNA expression induced by different concentrations of BMP4, indicating that BMP4 signaling pathway is not only involved in the process of cholinergic neuronal development, but also in the process of cholinergic neurons. The expression of homologous domain protein ISL-1 was also induced, which suggested that the extracellular signaling molecules of BMP4 might regulate the differentiation of terminal neural progenitor cells into cholinergic neurons by inducing homologous domain ISL-1 protein.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329.1

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