Transcriptional program coordination by N-methyl-D-aspartate-sensitive glutamate receptor stimulation in primary cultures of cerebellar neurons.
In primary cultures of rat cerebellar neurons, a brief stimulation of glutamate receptors results in coordinated activation of a programmed early gene response involving increases in the amount of c-fos, c-jun, jun-B, and zif/268 mRNAs. Each of these genes was induced to a different extent and showed a temporal pattern characterized by either a monophasic "early" response, occurring within 30 min of glutamate addition, or a biphasic response (c-jun), lasting for up to 6 to 8 hr after the initial stimulus. The early phase of the glutamate-induced gene expression was prevented by 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid, a highly selective isosteric antagonist of the N-methyl-D-aspartate (NMDA)-sensitive glutamate receptor (NMDA receptor). The second phase of the c-jun response was not blocked when the NMDA receptors were completely inhibited after the initial pulse of agonist or when the quisqualate-kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione was added, suggesting that a brief NMDA receptor stimulation triggers a cascade of events critical for the manifestation of the delayed c-jun expression. Furthermore, gel retardation assays demonstrated that NMDA receptor activation results in a prolonged increase in nuclear DNA-binding activity specific for the AP-1 transcriptional regulatory element. Protein immunoblot analysis showed that the composition of this nucleoprotein complex changes as a function of time, reflecting a cascade that involves an increased translation of Fos and several Fos-related proteins. The coordinated induction of several different transcription factors and the variations in transcriptional complex formation initiated by NMDA receptor stimulation may be a key mechanism in the orchestration of specific target gene expression that underlies various aspects of neuronal function, including plasticity responses.
Szekely
A M
AM
FIDIA-Georgetown Institute for the Neurosciences, Georgetown University, Washington, D.C. 20007.
Costa
E
E
Grayson
D R
DR
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D000818:Animals; D002478:Cells, Cultured / Q000187:drug effects; D002531:Cerebellum / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects*; D015151:Immunoblotting; D016202:N-Methylaspartate / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010622:Phencyclidine / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D011810:Quinoxalines / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D017470:Receptors, Glutamate; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D017981:Receptors, Neurotransmitter / Q000187:drug effects* / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D010879:Piperazines; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D011810:Quinoxalines; D017470:Receptors, Glutamate; D016194:Receptors, N-Methyl-D-Aspartate; D017981:Receptors, Neurotransmitter; D014157:Transcription Factors; D016202:N-Methylaspartate; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; C050592:3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid; D010622:Phencyclidine