title: Direct conversion of mouse fibroblasts to GABAergic neurons with combined medium without the introduction of transcription factors or miRNAs.
abstract: Degeneration or loss of GABAergic neurons frequently may lead to many neuropsychiatric disorders such as epilepsy and autism spectrum disorders. So far no clinically effective therapies can slow and halt the progression of these diseases. Cell-replacement therapy is a promising strategy for treatment of these neuropsychiatric diseases. Although increasing evidence showed that mammalian somatic cells can be directly converted into functional neurons using specific transcription factors or miRNAs via virus delivery, the application of these induced neurons is potentially problematic, due to integration of vectors into the host genome, which results in the disruption or dysfunction of nearby genes. Here, we show that mouse fibroblasts could be efficiently reprogrammed into GABAergic neurons in a combined medium composed of conditioned medium from neurotrophin-3 modified Olfactory Ensheathing Cells (NT3-OECs) plus SB431542, GDNF and RA. Following 3 weeks of induction, these cells derived from fibroblasts acquired the morphological and phenotypical GABAerigic neuronal properties, as demonstrated by the expression of neuronal markers including Tuj1, NeuN, Neurofilament-L, GABA, GABA receptors and GABA transporter 1. More importantly, these converted cells acquired neuronal functional properties such as synapse formation and increasing intracellular free calcium influx when treated with BayK, a specific activator of L-type calcium channel. Therefore, our findings demonstrate for the first time that fibroblasts can be directly converted into GABAergic neurons without ectopic expression of specific transcription factors or miRNA. This study may provide a promising cell source for the application of cell replacement therapy in neuropsychiatric disorders.
authors.affiliation: a State Key Laboratory of Oncogenes and Related Genes; Renji-MedX Clinical Stem Cell Research Center; Ren Ji Hospital; School of Medicine; Shanghai Jiao Tong University ; Shangha , China.
authors.forename: Huiming
authors.initials: H
authors.lastname: Xu
authors.forename: Yonghui
authors.initials: Y
authors.lastname: Wang
authors.forename: Zuping
authors.initials: Z
authors.lastname: He
authors.forename: Hao
authors.initials: H
authors.lastname: Yang
authors.forename: Wei-Qiang
authors.initials: WQ
authors.lastname: Gao
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000378:metabolism; D064987:Cell- and Tissue-Based Therapy / Q000379:methods; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000502:physiology*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004268:DNA-Binding Proteins; D004149:Dioxoles / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology*; D050485:GABA Plasma Membrane Transport Proteins / Q000096:biosynthesis; D059330:GABAergic Neurons / Q000166:cytology*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009422:Nervous System Diseases / Q000473:pathology / Q000628:therapy; D016900:Neurofilament Proteins / Q000096:biosynthesis; D055495:Neurogenesis / Q000502:physiology*; D009687:Nuclear Proteins / Q000096:biosynthesis; D018079:Receptors, GABA / Q000096:biosynthesis; D014404:Tubulin / Q000096:biosynthesis; D005680:gamma-Aminobutyric Acid / Q000096:biosynthesis
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D020746:Calcium Channels, L-Type; D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; D004149:Dioxoles; D050485:GABA Plasma Membrane Transport Proteins; D051100:Glial Cell Line-Derived Neurotrophic Factor; D035683:MicroRNAs; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D016900:Neurofilament Proteins; D009687:Nuclear Proteins; D018079:Receptors, GABA; D014404:Tubulin; C462941:beta3 tubulin, mouse; C044882:neurofilament protein L; C518911:neurotropin 3, mouse; D005680:gamma-Aminobutyric Acid
keywords: GABAergic neurons; combined medium; direct conversion; fibroblasts