mGluR5 is involved in dendrite differentiation and excitatory synaptic transmission in NTERA2 human embryonic carcinoma cell-derived neurons.
The pluripotent human embryonic carcinoma cell line NTERA2 readily differentiates into neurons when exposed to retinoic acid in vitro. These neurons show characteristic morphology with long processes and they express neuronal markers TUJ-1 and NeuN. NTERA2-derived neurons can regulate Ca2+ signalling through ionotropic glutamate (iGluR) and muscarinic receptors (mAChRs). Little is known, however, about the role of metabotropic glutamate receptors (mGluRs) in these neurons. Here we show that NTERA2-derived neurons express functional mGluR5, which is involved in Ca2+ signalling. Blocking mGluR5 activity at early stages of differentiation leads to fewer dendrites and a reduction in miniature excitatory postsynaptic currents (mEPSCs). Furthermore, cells cultured in the presence of the mGluR5 antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP) show reduced N-methyl-D-aspartate (NMDA) receptor-mediated Ca2+ mobilisation but increased alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor Ca2+ permeability. During normal neuronal development, the edited GluR2 renders AMPARs Ca2+ impermeable. The increased Ca2+ permeability of AMPARs in MPEP-treated neurons is due to the reduced expression of GluR2 subunit protein. Thus, mGluR5 activity at early stages of differentiation is likely to play a role in the development of multipotent cell-derived neurons.
Park
Hyeonseon
H
Biomedical Science, School of Medicine and Biomedical Science, University of Sheffield, Sheffield S10 2TN, UK.
Váradi
Anikó
A
Seok
Heon
H
Jo
Jihoon
J
Gilpin
Heather
H
Liew
Chee Gee
CG
Jung
Sungjun
S
Andrews
Peter W
PW
Molnár
Elek
E
Cho
Kwangwook
K
D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D045744:Cell Line, Tumor; D003712:Dendrites / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D004594:Electrophysiology; D018846:Excitatory Amino Acids / Q000502:physiology*; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D064529:Receptor, Metabotropic Glutamate 5; D018091:Receptors, AMPA / Q000096:biosynthesis / Q000187:drug effects; D018094:Receptors, Metabotropic Glutamate / Q000502:physiology*; D016194:Receptors, N-Methyl-D-Aspartate / Q000096:biosynthesis / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D009435:Synaptic Transmission / Q000502:physiology*
D018846:Excitatory Amino Acids; C578935:GRM5 protein, human; D064529:Receptor, Metabotropic Glutamate 5; D018091:Receptors, AMPA; D018094:Receptors, Metabotropic Glutamate; D016194:Receptors, N-Methyl-D-Aspartate; D002118:Calcium