title: Kainate exposure suppresses activation of GluR2 subunit promoter in primary cultured cerebral cortical neurons through induction of RE1-silencing transcription factor.
abstract: The AMPA receptor subunit GluR2 is downregulated in neurons following a wide range of neurological insults. Here we report that suppression of GluR2 gene promoter activity is associated with kainate (KA)-induced downregulation of GluR2 subunit levels in primary cultured cortical neurons. RT-PCR and Northern blotting showed a significant decrease in GluR2 mRNA in cultured neurons after KA exposure. Transfection of cultured neurons with an expression vector pGL3-GluR2(-298/+283), where the reporter gene firefly luciferase was driven by the GluR2 promoter, revealed that KA exposure suppressed the transcriptional activation of the GluR2 promoter. Furthermore, the expression of the RE1-silencing transcription factor (REST) was increased in KA-exposed cortical neurons; enhanced binding of REST to RE1-like silencer element in the proximal promoter of the GluR2 subunit gene was evidenced by electrophoresis mobility shift assay. Chromatin immunoprecipitation showed that suppressed activity of the GluR2 promoter in cultured neurons after KA exposure was related to deacetylation of histone H4. These results indicate that REST as a crucial factor binds to RE1-like silencer element in the GluR2 promoter, suppressing transcription of the GluR2 subunit gene during KA exposure. Our data suggest that transcriptional suppression of the GluR2 subunit gene may contribute at least in part to downregulation of GluR2 subunit protein in neurons during KA exposure. Because our experiments showed a reduction of glutamate release in KA-exposed cortical neurons, REST may play a latent role in delayed neuronal death or in seizure-induced tolerance.
authors.affiliation: Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Xue Yuan Road 38, Beijing 100083, PR China.
authors.forename: Yu-Hong
authors.initials: YH
authors.lastname: Jia
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhu
authors.forename: Shu-Yan
authors.initials: SY
authors.lastname: Li
authors.forename: Ju-Hua
authors.initials: JH
authors.lastname: Ni
authors.forename: Hong-Ti
authors.initials: HT
authors.lastname: Jia
mesh_terms: D000107:Acetylation; D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D015536:Down-Regulation; D024202:Electrophoretic Mobility Shift Assay; D017930:Genes, Reporter; D006657:Histones / Q000378:metabolism; D007608:Kainic Acid / Q000378:metabolism* / Q000633:toxicity; D049409:Luciferases, Firefly / Q000037:antagonists & inhibitors / Q000235:genetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000235:genetics; D012333:RNA, Messenger / Q000037:antagonists & inhibitors; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000037:antagonists & inhibitors* / Q000235:genetics; D012097:Repressor Proteins / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D037941:Silencer Elements, Transcriptional; D014157:Transcription Factors / Q000096:biosynthesis*; D000099229:RE1-Silencing Transcription Factor
chemical_list: D006657:Histones; D007608:Kainic Acid; D049409:Luciferases, Firefly; D021122:Protein Subunits; D012333:RNA, Messenger; D018091:Receptors, AMPA; D012097:Repressor Proteins; D014157:Transcription Factors; D000099229:RE1-Silencing Transcription Factor; C104722:glutamate receptor ionotropic, AMPA 2