摘要
采用BDNF、GDNF、PDGF和forskolin对培养不同时间的大鼠脑神经干细胞(NSC)进行诱导分化,通过免疫荧光染色观察这些诱导剂对NSC分化为神经元的作用,以RTPCR和Westernblot检测体外培养不同时间的NSC表达TrkB受体的水平,探讨它与NSC诱导分化反应性的关系。结果表明,各种神经营养因子诱导的MAP2阳性细胞数均高于未加神经营养因子的对照组(P<0.001)。在所使用的神经营养因子中,以BDNF促进NSC分化成神经元的效果为著,MAP2阳性细胞数约为空白对照组的6倍。而体外培养3个月的NSC单用BDNF诱导分化后未见MAP2阳性细胞,仅在联合使用BDNF和forskolin时方可诱导出少数MAP2阳性细胞。RTPCR和Westernblot检测结果显示,体外培养3个月的NSC表达TrkB受体的mRNA及蛋白的量较培养1个月的NSC减少3倍,提示培养3个月的NSC对BDNF诱导反应性的减弱可能与TrkB受体表达下调有关。
Neuronal induction on NSCs using BDNF, GDNF, PDGF and forskolin was examined by immunofluoresence staining. The change of TrkB receptor expression in cultured NSCs at different culture time was detected by RT-PCR and Western blot assay. The results showed that the percentage of MAP2 positive cells in l-month-old NSCs increased about 6 times compared to the control after neumtrophic factors induction （ P 〈0. 001 ）, and BDNF was the most effective one among them. Only a few MAP2-positive cells were seen in 3-monthold NSCs by combination of BDNF and forskolin treatment. The resuhs of RT-PCR and Western blot analysis showed that the expression of the TrkB receptor in 3-month-old NSCs decreased 3 times compared to that of 1-month-old NSCs. The present results suggest that reduction of response to BDNF of NSCs cultured for 3 months may relate to the decline of TrkB receptor expression.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2006年第2期214-218,共5页
Chinese Journal of Neuroanatomy
基金
福建省科技计划项目(2001Z020)
福建省自然科学基金(C0310022)资助项目
作者简介
林玲：通讯作者 电话：0591-83569300 E-mail：llinzheng@163.com
