title: Neurotrophin-3 promotes proliferation and cholinergic neuronal differentiation of bone marrow- derived neural stem cells via notch signaling pathway.
abstract: OBJECTIVE
  Recently, the potential for neural stem cells (NSCs) to be used in the treatment of Alzheimer's disease (AD) has been reported; however, the therapeutic effects are modest by virtue of the low neural differentiation rate. In our study, we transfected bone marrow-derived NSCs (BM-NSCs) with Neurotrophin-3 (NT-3), a superactive neurotrophic factor that promotes neuronal survival, differentiation, and migration of neuronal cells, to investigate the effects of NT-3 gene overexpression on the proliferation and differentiation into cholinergic neuron of BM-NSCs in vitro and its possible molecular mechanism.
  
  
  METHODS
  BM-NSCs were generated from BM mesenchymal cells of adult C57BL/6 mice and cultured in vitro. After transfected with NT-3 gene, immunofluorescence and RT-PCR method were used to determine the ability of BM-NSCs on proliferation and differentiation into cholinergic neuron; Acetylcholine Assay Kit was used for acetylcholine (Ach). RT-PCR and WB analysis were used to characterize mRNA and protein level related to the Notch signaling pathway.
  
  
  RESULTS
  We found that NT-3 can promote the proliferation and differentiation of BM-NSCs into cholinergic neurons and elevate the levels of acetylcholine (ACh) in the supernatant. Furthermore, NT-3 gene overexpression increase the expression of Hes1, decreased the expression of Mash1 and Ngn1 during proliferation of BM-NSCs. Whereas, the expression of Hes1 was down-regulated, and Mash1 and Ngn1 expression were up-regulated during differentiation of BM-NSCs.
  
  
  CONCLUSIONS
  Our findings support the prospect of using NT-3-transduced BM-NSCs in developing therapies for AD due to their equivalent therapeutic potential as subventricular zone-derived NSCs (SVZ-NSCs), greater accessibility, and autogenous attributes.
authors.affiliation: School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, PR China.
authors.forename: Yu-Hui
authors.initials: YH
authors.lastname: Yan
authors.affiliation: School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, PR China.
authors.forename: Shao-Heng
authors.initials: SH
authors.lastname: Li
authors.affiliation: Department of Interventional Therapy, Department of Rehabilitation, Dalian Municipal Central Hospital, Dalian 116033, PR China.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Department of Interventional Therapy, Department of Rehabilitation, Dalian Municipal Central Hospital, Dalian 116033, PR China.
authors.forename: Sa-Feng
authors.initials: SF
authors.lastname: Zou
authors.affiliation: School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, PR China.
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Li
authors.affiliation: School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, PR China.
authors.forename: Zhen-Yu
authors.initials: ZY
authors.lastname: Tao
authors.affiliation: School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Song
authors.affiliation: School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, PR China. Electronic address: jingxianyang0520@163.com.
authors.forename: Jing-Xian
authors.initials: JX
authors.lastname: Yang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D059329:Cholinergic Neurons / Q000166:cytology* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D055495:Neurogenesis; D020933:Neurotrophin 3 / Q000235:genetics / Q000378:metabolism*; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction; D014161:Transduction, Genetic; D015854:Up-Regulation
chemical_list: D020933:Neurotrophin 3; D051880:Receptors, Notch
keywords: Alzheimer's disease; Bone marrow-derived neural stem cells; Cholinergic neurons; Neurotrophin-3; Notch signaling pathway; Proliferation