胶质细胞源性神经营养因子对培养的胚鼠脑室下区神经干细胞表达Pitx3及酪氨酸羟化酶的影响

Effect of glial cell line-derived neurotrophic factors on expression of Pitx3 and tyrosine hydroxyase of cultured neural stem cells subventricular zone in rat brain

目的:观察胶质细胞源性神经营养因子(GDNF)对培养的胚鼠脑室下区(SVZ)神经干细胞(NSCs)表达Pitx3、酪氨酸羟化酶(TH)的影响。方法:取胚鼠SVZ组织,进行细胞培养。分为GDNF+NSCs共培养组和NSCs单独培养对照组。用Pitx3、TH、Pitx3/TH免疫荧光单标和双标及Nestin免疫组化法进行检测。结果:共培养组5 d时形成神经球,10 d时球体积不断增大,12 d后开始贴壁生长,并向四周伸出突起并开始分化。对照组神经球明显少于共培养组。14 d共培养组Pitx3、TH、Pitx3/TH阳性细胞明显高于对照组。结论:GDNF能上调SVZ区NSCs表达Pitx3和TH,诱导NSCs向多巴胺(DA)能神经元分化。

Objective： To investigate the effect of glial cell line-derived neurotrophic factors （GDNF） on expression of Pitx3 and TH in neural stem cells （NSCs） of subventricular zone （SVZ） in rat brain. Methods： NSCs were separated from SVZ of ED14 rat and cultured. The cells were randomly divided into two groups：NSCs co-cultured with GDNF and NSCs cultured solely as the control. The proliferation and differentiation of the neural stem cells were respectively identified using Nestin stained immunohistochemistry and Pitx3, TH single or double-stained immunofluorescence. Results： The co-cultured groups grew to form typical floaty neurospheres at 5 d after seeding. At 12 d, the neurospheres in co-cuhrured and NSCs sole cuhrured groups were 12.40±3. 03, 9.20±2.97（P〈0. 01）respectively. （）n the 14th day, Pitx3, TH and Pitx3/TH positive cells in the co-cultured group（8.60%, 3.09% and 2.21%, respectively）were significantly more than those in the control group（0. 67%, 0. 76% and 0.48%, respectively）（P〈0. 01）. Conclusion： GDNF can increase the expression of Pitx3 and TH of NSCs in SVZ, and induce differentiation of NSCs to dopaminergic neuron.