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Title: 神经营养因子一3与成纤维生长因子一2在神经前体细胞的增殖和分化中的相互作用及其机制研究
Author: 金璐
Degree Level: 博士
Issued Date: 2005
Degree Grantor: 中国科学院神经科学研究所
Place of Degree Grantor: 中国科学院神经科学研究所
Supervisor: 冯林音
Keyword: 神经干细胞 ; 碱性成纤维生长因子-2(FGF2) ; 神经营养因子-3(NT3) ; 抑制作用 ; 增殖 ; 少突胶质细胞 ; 分化
Alternative Title: Interaction between FGF2 and NT3 on Neural Stem Cell Proliferation and Differentiation
Abstract: 神经干细胞是一类具有自我增殖能力和多分化潜能的细胞,其增殖和分化受各类因子的调控。神经干细胞的增殖和分化是个极其复杂的过程,迄今为止,因子对这一过程调控的详细机制还不清楚,因此,研究因子在神经干细胞增殖分化中的的调控作用,尤其是多个因子的一协同调控作用及其机制,对于深入了解神经干细胞在体内的发育和功能具有重要意义。本研究应用体外培养的新生大鼠海马区神经前体细胞进行多个因子的协同作用的的研究,发现神经营养因子-3(NT3)和碱性成纤维生一氏因子-2(FGF2)共同作用于神经干细胞时,NT3可抑制FGF2引起的神经细胞球生长和澳脱氧尿普(BrdU)的掺入。抑制MEK和P13K信号传导通路都能抑制FGF2引起的BrdU的掺入,说明这两条通路都为FGF2引起神经前体细胞增殖所必需。进一步研究表明NT3抑制了FGFZ引起的Akt及其下游分子GSK3p的磷酸化,而对ERK1/2的磷酸化水平没有影响。NT3对FGF2依赖的神经干细胞增殖的抑制作用可被PI3K抑制剂LY294002所阻断。GSK3的抑制剂SB216763也能阻断NT3的抑制作用,恢复神经细胞球的生长和BrdU的掺入。抑制内源性NT3可促进FGF2引起的神经干细胞的增殖;FGF2存在的情况下,抗NT3血清的加入使Akt/GSK3β的磷酸化水平升高。本研究结果表明:NT3通过P13幻GSK3日信号通路抑制FGF2在神经前体细胞中的增殖作用。同时,本研究还考察了NT3与FGF2在神经前体细胞分化中的相互作用。来源于生后海马区域的神经前体细胞具有多潜能性,能分化为神经元,星状胶质细胞和少突胶质细胞。利用少突胶质细胞特异表达蛋白POGFR以和脂类分子O4的抗体进行免疫细胞染色分析,结果表明,单独加入NT3或FGF2都可促进神经前体细胞向少突胶质细胞方向分化,同时加入NT3和FGF2,少突胶质细胞的分化比例反呈下降趋势,说明NT3和FGF2在少突胶质细胞的分化上存在拮抗作用。我们的研究表明,多种因子对神经前体细胞增殖和分化的相互作用是一个复杂的过程。
English Abstract: During development, NP cells in the hippocampus are exposed to a variety of cues with different effects, which are integrated by NP cells so that an appropriate number of cells are ultimately generated. It is therefore important to understand the antagonism and cooperation between cellular factors and the mechanism underlying. In this dissertation, using an FGF2-dependent rat neurosphere culture system, we found that NT3 inhibited both FGF2-induced neurosphere growth and bromodeoxyuridine (BrdU) incorporation in a dose-dependent manner. U0126, a MEK1/2 inhibitor, and LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, both inhibited BrdU incorporation induced by FGF2, suggesting that the extracellular signal-regulated kinasel/2 (ERK1/2) and PI3K pathways are required for FGF2-induced neural progenitor cell proliferation. We found that NT3 significantly inhibited FGF2-induced phosphoryiation of Akt and glycogen synthase kinase3p (GSK3(3), a downstream kinase of Akt, whereas NT3 did not affect phosphoryiation of ERK1/2. The inhibitory effect of NT3 on FGF2-induced neural progenitor cell proliferation was abolished by LY294002 and treatment of SB216763, a specific glycogen synthase kinase 3 (GSK3) inhibitor, prevented the inhibitory effect of NT3 on FGF2-induced NP cell proliferation and rescued both neurosphere growth and BrdU incorporation. Moreover, experiments with Anti-NT3 antibody showed that endogenous NT3 also played a role in inhibiting FGF2-induced NP cell proliferation and that the Anti-NT3 antibody enhanced phospho-Akt and phospho-GSK3(3 levels in the presence of FGF2. These findings indicate that FGF2-induced NP cell proliferation is inhibited by NT3 via the PI3K /GSK3 pathway. The second set of experiment is to study the role of NT3 and FGF2 in the differentiation of neural stem cell. Under appripriate conditions, multipotential neural stem cell can differentiate to neuron, astrocyte, or oligodendrocyte. Experiment results show that the addition of NT3 or FGF2 alone induces neural stem cell to differentiate to oligodendrocyte. While the ratio of oligodendrocyte decreased to the base level when NT3 and FGF2 were added together. Thus an antagonism exits between the actions of NT3 and FGF2 on oligodendrocyte differentiation.
Language: 中文
Content Type: 学位论文
URI: http://ir.sibs.ac.cn/handle/331001/2262
Appears in Collections:神经所(总)_学位论文

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Recommended Citation:
神经营养因子一3与成纤维生长因子一2在神经前体细胞的增殖和分化中的相互作用及其机制研究.金璐[d].中国科学院神经科学研究所,2005.20-25
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