Abstract
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 α-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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 482-489 |
| Number of pages | 8 |
| Journal | Journal of Neurochemistry |
| Volume | 63 |
| Issue number | 2 |
| State | Published - Aug 1994 |
| Externally published | Yes |
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Keywords
- α-Amino-3- hydroxy-5-methyl-4-isoxazolepropionate
- Cell culture
- Excitotoxicity
- Glutamate receptor channels
- Kainate
- Neurons
- Patch clamp
- Reverse transcriptase- polymerase chain reaction
ASJC Scopus subject areas
- Biochemistry
- Cellular and Molecular Neuroscience
Cite this
Expression of non-NMDA glutamate receptor channel genes by clonal human neurons. / Hardy, Mattie; Younkin, Donald; Tang, Cha Min; Pleasure, Jeanette; Shi, Qing Yao; Williams, Margaret; Pleasure, David E.
In: Journal of Neurochemistry, Vol. 63, No. 2, 08.1994, p. 482-489.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Expression of non-NMDA glutamate receptor channel genes by clonal human neurons
AU - Hardy, Mattie
AU - Younkin, Donald
AU - Tang, Cha Min
AU - Pleasure, Jeanette
AU - Shi, Qing Yao
AU - Williams, Margaret
AU - Pleasure, David E
PY - 1994/8
Y1 - 1994/8
N2 - 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 α-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.
AB - 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 α-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.
KW - α-Amino-3- hydroxy-5-methyl-4-isoxazolepropionate
KW - Cell culture
KW - Excitotoxicity
KW - Glutamate receptor channels
KW - Kainate
KW - Neurons
KW - Patch clamp
KW - Reverse transcriptase- polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=0028133258&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028133258&partnerID=8YFLogxK
M3 - Article
C2 - 7518497
AN - SCOPUS:0028133258
VL - 63
SP - 482
EP - 489
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
SN - 0022-3042
IS - 2
ER -