title: Fyn Kinase regulates GluN2B subunit-dominant NMDA receptors in human induced pluripotent stem cell-derived neurons.
abstract: NMDA receptor (NMDAR)-mediated fast excitatory neurotransmission is implicated in a broad range of physiological and pathological processes in the mammalian central nervous system. The function and regulation of NMDARs have been extensively studied in neurons from rodents and other non-human species, and in recombinant expression systems. Here, we investigated human NMDARs in situ by using neurons produced by directed differentiation of human induced pluripotent stem cells (iPSCs). The resultant cells showed electrophysiological characteristics demonstrating that they are bona fide neurons. In particular, human iPSC-derived neurons expressed functional ligand-gated ion channels, including NMDARs, AMPA receptors, GABAA receptors, as well as glycine receptors. Pharmacological and electrophysiological properties of NMDAR-mediated currents indicated that these were dominated by receptors containing GluN2B subunits. The NMDAR currents were suppressed by genistein, a broad-spectrum tyrosine kinase inhibitor. The NMDAR currents were also inhibited by a Fyn-interfering peptide, Fyn(39-57), but not a Src-interfering peptide, Src(40-58). Together, these findings are the first evidence that tyrosine phosphorylation regulates the function of NMDARs in human iPSC-derived neurons. Our findings provide a basis for utilizing human iPSC-derived neurons in screening for drugs targeting NMDARs in neurological disorders.
authors.affiliation: Program in Neurosciences &Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.|Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Wen-Bo
authors.initials: WB
authors.lastname: Zhang
authors.affiliation: Program in Developmental &Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.
authors.forename: P Joel
authors.initials: PJ
authors.lastname: Ross
authors.affiliation: Program in Neurosciences &Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.
authors.forename: YuShan
authors.initials: Y
authors.lastname: Tu
authors.affiliation: Program in Neurosciences &Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.
authors.forename: Yongqian
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Program in Neurosciences &Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.|Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Simon
authors.initials: S
authors.lastname: Beggs
authors.affiliation: Program in Neurosciences &Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.
authors.forename: Ameet S
authors.initials: AS
authors.lastname: Sengar
authors.affiliation: Program in Developmental &Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.|Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
authors.forename: James
authors.initials: J
authors.lastname: Ellis
authors.affiliation: Program in Neurosciences &Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A4, Canada.|Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Salter
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D009473:Neuronal Plasticity; D009474:Neurons / Q000378:metabolism / Q000502:physiology*; D051076:Proto-Oncogene Proteins c-fyn / Q000378:metabolism*; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D009435:Synaptic Transmission / Q000187:drug effects
chemical_list: C121001:NR2B NMDA receptor; D016194:Receptors, N-Methyl-D-Aspartate; D019833:Genistein; C494957:FYN protein, human; D051076:Proto-Oncogene Proteins c-fyn