Expression of non-NMDA glutamate receptor channel genes by clonal human neurons.
Treatment of the human teratocarcinoma line NTera2/c1.D1 (NT2) with retinoic acid induces terminal neuronal differentiation. In a previous study, we found that the neurons obtained in this way express functional N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptor channels. We now show by reverse transcriptase-polymerase chain reaction and Southern blotting that these neurons transcribe each of the nine known non-NMDA glutamate receptor genes (GluR1-7, Ka-1, and Ka-2) and that four of these genes (GluR2, GluR6, GluR7, and Ka-1) are also transcribed by undifferentiated NT2 cells. Patch clamp studies demonstrate that individual non-NMDA glutamate receptor channels are readily isolated from NT2-derived neurons and that these channels are potently modulated by the desensitization blocker cyclothiazide. NT2-derived neurons are susceptible to kainate excitotoxicity but are not injured by prolonged exposure to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate. We expect that the NT2-derived human neuronal culture system will facilitate studies of human neuronal non-NMDA glutamate receptor channels and of the pathophysiology of neuronal excitotoxicity.
Hardy
M
M
Division of Neurology Research, Children's Hospital of Philadelphia, PA 19104.
Younkin
D
D
Tang
C M
CM
Pleasure
J
J
Shi
Q Y
QY
Williams
M
M
Pleasure
D
D
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D001483:Base Sequence; D015139:Blotting, Southern; D002454:Cell Differentiation / Q000187:drug effects; D002999:Clone Cells; D017931:DNA Primers; D015870:Gene Expression; D006801:Humans; D007473:Ion Channels / Q000096:biosynthesis* / Q000502:physiology; D007608:Kainic Acid / Q000633:toxicity; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011810:Quinoxalines / Q000494:pharmacology; D017470:Receptors, Glutamate / Q000096:biosynthesis*; D018243:Teratocarcinoma; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000633:toxicity
D017931:DNA Primers; D007473:Ion Channels; D011810:Quinoxalines; D017470:Receptors, Glutamate; D014212:Tretinoin; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D007770:L-Lactate Dehydrogenase; D007608:Kainic Acid