Glutamate receptor-mediated currents and toxicity in embryonal carcinoma cells.
While primary neuronal cell cultures have been used to investigate excitotoxicity, development of cell lines exhibiting glutamate receptor-mediated death is desirable. P19 mouse embryonal carcinoma cells, exposed to retinoic acid and plated onto a layer of cultured mouse cortical glial cells, differentiated into neuron-like elements immunoreactive for neurofilaments and neuron-specific enolase. Whole-cell recordings revealed inward currents in response to extracellular application of either NMDA or kainate. The NMDA-induced currents exhibited a voltage-dependent blockade by magnesium, required glycine for maximal activation, and were blocked by the NMDA antagonist dizocilpine. Kainate-induced currents were blocked by the AMPA/kainate receptor antagonist CNQX. Exposure to 500 microM NMDA for 24 h destroyed most P19 cells (EC50 approximately 70 microM); death was prevented by dizocilpine or D-APV. Exposure to 500 microM kainate also resulted in widespread death reduced by CNQX. Thus differentiated P19 cells exhibited both excitatory amino acid responses and vulnerability to excitotoxicity, characteristic of CNS neurons. These cells may provide a genetically open system useful for studying glutamate receptor-mediated phenomena at a molecular level.
Turetsky
D M
DM
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri 63110.
Huettner
J E
JE
Gottlieb
D I
DI
Goldberg
M P
MP
Choi
D W
DW
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D000596:Amino Acids / Q000633:toxicity; D000818:Animals; D018236:Carcinoma, Embryonal / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004594:Electrophysiology; D005260:Female; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D007473:Ion Channels / Q000378:metabolism; D007608:Kainic Acid / Q000633:toxicity; D051379:Mice; D016202:N-Methylaspartate / Q000633:toxicity; D009457:Neuroglia / Q000502:physiology; D009474:Neurons / Q000378:metabolism*; D011247:Pregnancy; D011810:Quinoxalines / Q000494:pharmacology; D017470:Receptors, Glutamate / Q000187:drug effects / Q000502:physiology*; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors; D014212:Tretinoin / Q000633:toxicity; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000596:Amino Acids; D007202:Indicators and Reagents; D007473:Ion Channels; D011810:Quinoxalines; D017470:Receptors, Glutamate; D016194:Receptors, N-Methyl-D-Aspartate; D014212:Tretinoin; D016202:N-Methylaspartate; D016291:Dizocilpine Maleate; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D007608:Kainic Acid