DO SEIZURES DISRUPT CORTICAL GENE EXPRESSION

癫痫发作会破坏皮质基因表达吗

• 批准号: 2703053
• 负责人: KENNETH J MACK
• 金额: $ 17.42万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2703053
• 关键词: calcineurin; cerebral cortex; gel mobility shift assay; gene induction /repression; genetic promoter element; kindling; laboratory rat; messenger RNA; nerve /myelin protein; neural plasticity; neurogenetics; neurons; neurotrophic factors; polymerase chain reaction; protein structure function; sensory mechanism; synapsins; synaptic vesicles; tissue /cell culture; transcription factor; transfection; transmission electron microscopy; western blottings

Data from this lab has demonstrated that sensory experience can produce changes in the expression of transcriptional regulatory factors (such as NGFI-A) and their potential target genes (such as glutamic acid decarboxylase (67 kD form) and synaptophysin) in the barrel cortex. This system serves as a model to study how sensory experience can effect plasticity in neuronal cells, particularly since many of the identified target genes for NGFI-A have a functional role at the synapse. In contrast to a physiological stimulus, this lab has shown that seizures inhibit the transcriptional activation of these synaptic target genes. Based on these preliminary studies, this proposal seeks to test the hypothesis that seizures will interfere with the physiological transcriptional activation that occurs in the cortex after sensory experience, and therefore may interfere with the plasticity that occurs after sensory experience. Predictions based on this hypothesis will be tested by studying animals exposed to both physiological and seizure stimuli. Gene expression will be monitored by a combination of mRNA (RT-PCR and in situ hybridization) and protein (Western blot and immunohistochemical) techniques. The interactions of the transcription factors with their target genes will be addressed using gel shift analysis and transient co-transfection assays. Long term measures of synaptic plasticity will be studied at both the light microscopic and electron microscopic level. The results of these studies extend our current understanding of both the changes in transcriptional activity that occur after physiological neuronal stimulation, as well as the effects of seizure activity on these pathways.

该实验室的数据表明，感官体验可以产生 转录调节因素表达的变化（例如 NGFI-A）及其潜在靶基因（例如谷氨酸 枪管皮层中的脱羧酶（67 kD形式）和突触素）。这 系统是研究感官体验如何影响的模型 神经元细胞中的可塑性，特别是因为许多已识别 NGFI-A的靶基因在突触中具有功能作用。在 与生理刺激形成对比，该实验室表明癫痫发作 抑制这些突触靶基因的转录激活。 基于这些初步研究，该建议旨在测试 假设癫痫发作会干扰生理 感官后在皮层中发生的转录激活 经验，因此可能会干扰发生的可塑性 经过感官经验。 基于此假设的预测将通过研究动物来检验 暴露于生理和癫痫发作刺激。基因表达会 通过mRNA（RT-PCR和原位杂交）的组合来监测 和蛋白质（蛋白质印迹和免疫组织化学）技术。这 转录因子与目标基因的相互作用将 使用凝胶移位分析和瞬态共转染来解决 测定。将研究突触可塑性的长期测量 光显微镜和电子显微镜水平。 这些研究的结果扩展了我们目前对 生理后发生的转录活性的变化 神经元刺激以及癫痫活性对这些刺激的影响 途径。