Fyn Kinase regulates GluN2B subunit-dominant NMDA receptors in human induced pluripotent stem cell-derived neurons.

Wen-Bo Zhang,James Ellis,YuShan Tu,Yongqian Wang,Michael W. Salter,P. Joel Ross,Simon Beggs,Ameet S. Sengar

doi:10.1038/srep23837

Wen-Bo Zhang, James Ellis + Show 6 more

Open Access

https://doi.org/10.1038/srep23837

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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.

Highlights

NMDA receptors (NMDARs) are a prominent subtype of ionotropic glutamate receptor mediating fast excitatory synaptic transmission in the central nervous system (CNS)[1]
We found that GluN2B subunit-containing NMDARs were dominant and that tyrosine-protein kinase Fyn potentiated the function of GluN2B subunit-containing NMDARs in human Induced pluripotent stem cells (iPSCs)-derived neurons
We investigated the properties of human NMDARs in situ in iPSC-derived neurons

Summary

Introduction

NMDA receptors (NMDARs) are a prominent subtype of ionotropic glutamate receptor mediating fast excitatory synaptic transmission in the central nervous system (CNS)[1]. The GluN2 subunits have a key role in determining NMDAR properties, such as receptor gating, pharmacology, Ca2+ permeability, and blockade by extracellular Mg2+, leading to heterogeneity in NMDAR function across the CNS. From studies using non-human neurons, it is well-known that the function of NMDARs is regulated by members of the Src family of non-receptor protein tyrosine kinases[5,6]. Src family kinases contribute to physiological and pathological changes in synaptic strength by modulating NMDAR function and are involved in brain development, learning and memory formation, and pathogenesis of neurological disorders, such as chronic pain and schizophrenia[7,8]. In the present study we investigated the function and regulation of human NMDARs using human iPSC-derived neurons. We found that GluN2B subunit-containing NMDARs were dominant and that tyrosine-protein kinase Fyn potentiated the function of GluN2B subunit-containing NMDARs in human iPSC-derived neurons

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