摘要
目的研究Desert Hedgehog(DHH)对胚胎中脑神经前体细胞(NPCs)向多巴胺能神经元分化的诱导作用。方法大鼠DHH病毒上清感染COS7、NIH/3T3和9L细胞,用免疫荧光、实时定量PCR和Western blot方法检测DHH表达。分离培养大鼠胚胎中脑神经前体细胞,免疫荧光鉴定神经前体细胞的增殖分化特性。条件细胞表达DHH成功后分别与上述NPCs共培养10 d,免疫荧光检测酪氨酸羟化酶(TH)的表达。结果DHH在COS7、NIH/3T3和9L细胞中成功表达;同时,分离培养的大鼠胚胎中脑神经前体细胞具有良好的增殖分化特性,但两者共培养10 d后偶见TH阳性的细胞。结论DHH编码的蛋白不能或不能单独诱导NPCs定向分化为多巴胺能神经元。
Objective To investigate the effect of Desert Hedgehog on direct differentiation of neural progenitor cells （NPCs） cultured from embryonic mesencephalon in the rats. Methods We infected DHH into COS7, NIH/ 3T3 and 9L cells, and detected the expression of DHH in the cells with immunofluorescence, real-time PCR and Western blot. All of the three cells were co-cultured with NPCs isolated from ventral mesencephalon in embryonic SD rats （El3.5） respectively. Immunoreactivities of tyrosine hydroxylase （TH） was detected by immunocytochemistry after 10 days. Results The expression of DHH in COS7, NIH/3T3 and 9L cells was remarkably detected, but few TH-positive cells were found in the three co-cultral systems at the 10th day. Conclusion The protein derived from DHH itself does not show any inductive effect on the differentiation of NPCs to the dopaminergic neurons in vitro.
出处
《基础医学与临床》
CSCD
北大核心
2009年第4期360-365,共6页
Basic and Clinical Medicine
基金
国家重点基础研究发展计划(973)(2007CB947704)
