Neurotrophin 3 gene overexpression promotes cholinergic differentiation in neural stem cells
10.3969/j.issn.1001-1978.2016.05.009
- VernacularTitle:NT-3高表达促进神经干细胞向胆碱能神经元分化
- Author:
Yuhui YAN
;
Shaoheng LI
;
Liang KONG
;
Yingjia YAO
;
Yanan JIAO
;
Zhenyu TAO
;
Jie SONG
;
Jingxian YANG
- Publication Type:Journal Article
- Keywords:
neural stem cells;
Alzheimer ’ s disease;
neurotrophin-3;
cholinergic neuron;
differentiation;
Notch signaling pathway
- From:
Chinese Pharmacological Bulletin
2016;32(5):631-637
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the effects of neurotro-phin-3 ( NT-3 ) gene overexpression on the differentia-tion into cholinergic neuron of neural stem cells ( NSCs) in vitro and its underlying mechanism. Meth-ods Brain-derived NSCs from newborn mice were iso-lated and cultured in vitro and determined by immuno-fluorescence. The NSCs were divided into three groups: NSCs, GFP-NSCs and NT-3-NSCs groups. The expression of NT-3 was detected by immunofluo-rescence and ELISA. Then, the ability of NSCs on dif-ferentiation into cholinergic neuron was detected by im-munofluorescence and RT-PCR, and the Acetylcholine Assay Kit was used for acetylcholine ( ACh) , and the expression of Hes1 , Mash1 and Ngn1 mRNA was de-termined by RT-PCR. Results The neurosphere dis-played Nestin and Sox 2-postive by immunofluores-cence, suggesting that the cultured cells were NSCs. The proportion of ChAT immunopositive cells was sig-nificantly higher in the NT-3-NSCs group than that in the other two groups ( P <0. 01 ) . Ach secretion in NT-3-NSCs was significantly elevated compared with the other two groups ( P <0. 01 ) . NSCs transfected with NT-3 increased the levels of Mash1 and Ngn1 mR-NA, and decreased the level of Hes1 mRNA ( P <0. 05 ) . Conclusion NT-3 can significantly promote the in vitro differentiation of NSCs into cholinergic neu-rons via probablly inhibiting Notch signaling pathway.
