title: Over-expression of Mash1 improves the GABAergic differentiation of bone marrow mesenchymal stem cells in vitro.
abstract: Bone marrow mesenchymal stem cells (BMSCs) have been shown to be a promising cell type for the study of neuronal differentiation; however, few attempts had been made to differentiate these cells into inhibitory gamma-aminobutyric acid (GABA)ergic neurons. In this study, we over-expressed mammalian achaete-scute homologue-1 (Mash1), a basic helix-loop-helix (bHLH) transcription factor, in Sprague-Dawley rat BMSCs via lentiviral vectors, and then induced neuronal differentiation of these cells using conditioned medium. Our Western blot results show that, under conditions of differentiation, Mash1-overexpressing BMSCs exhibit an increased expression of neuronal markers and a greater degree of neuronal morphology compared to control, non-Mash1-overexpressing cells. Using immunocytochemistry, we observed increased expression of glutamic acid decarboxylase 67 (GAD67), as well as neuron-specific nuclear protein (NeuN) and β3-tubulin, in Mash1-overexpressing BMSCs compared to control cells. Moreover, we also found the differentiated cells showed representative traces of action potentials in electrophysiological characterization. In conclusion, our study demonstrated that over-expression of Mash1 can improve GABAergic differentiation of BMSCs in vitro.
authors.affiliation: Department of Neurosurgery, Xijing Institute of Clinical Neuroscience, Fourth Military Medical University, No.17 Chang-le West Road, Xi'an 710032, China; Department of Neurosurgery, Qingdao 401 Hospital of PLA, No. 22 Minjiang Road, Qingdao 266071, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.forename: Qianfa
authors.initials: Q
authors.lastname: Long
authors.forename: Chenguang
authors.initials: C
authors.lastname: Jia
authors.forename: Yuanyang
authors.initials: Y
authors.lastname: Liu
authors.forename: Xicai
authors.initials: X
authors.lastname: Yi
authors.forename: Hao
authors.initials: H
authors.lastname: Yang
authors.forename: Zhou
authors.initials: Z
authors.lastname: Fei
authors.forename: Weiping
authors.initials: W
authors.lastname: Liu
mesh_terms: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D002452:Cell Count; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D058787:GABA-A Receptor Antagonists / Q000494:pharmacology; D059330:GABAergic Neurons / Q000502:physiology*; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000378:metabolism; D010852:Picrotoxin / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D054830:Sesterterpenes; D014633:Valine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D051792:Basic Helix-Loop-Helix Proteins; D018691:Excitatory Amino Acid Antagonists; D058787:GABA-A Receptor Antagonists; D005968:Glutamate Decarboxylase; D010751:Phosphopyruvate Hydratase; D010852:Picrotoxin; D054830:Sesterterpenes; D014633:Valine; C499318:Ascl1 protein, rat; C056639:2-amino-5-phosphopentanoic acid; C022961:picrotoxinin; C402565:glutamate decarboxylase 1
keywords: BMSCs; Bone marrow mesenchymal stem cell; Differentiation; GABA; GAD67; Mash1; NeuN; bHLH; basic helix-loop-helix; bone marrow mesenchymal stem cells; gamma-aminobutyric acid; glutamic acid decarboxylase 67; mammalian achaete-scute homologue-1; neuron-specific nuclear protein