ID of Drosophila proteins modified by neural activity

神经活动修饰的果蝇蛋白质的 ID

• 批准号: 6508673
• 负责人: MANI RAMASWAMI
• 金额: $ 15.15万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6508673
• 关键词: Drosophilidae; antibody; arthropod genetics; biological signal transduction; enzyme activity; gel electrophoresis; gene expression; mass spectrometry; mitogen activated protein kinase; neural plasticity; neurons; polymerase chain reaction; posttranscriptional RNA processing; posttranslational modifications; protein sequence; transcription factor; ubiquitin; western blottings

DESCRIPTION: (provided by applicant) Experience-dependent neural plasticity underlies drug addiction, complex, enduring behavioral changes; that result after drug use. Molecules rapidly regulated by neural activity are likely to be very important for initiating plasticity processes. Thus, their identification and characterization is of particular relevance for understanding cellular mechanisms that underlie behavioral change. This proposal seeks to combine novel methodologies to identify molecules subject to rapid, post-transcriptional regulation during establishment of long-term synaptic change. Recent technical advances made in the principal investigator's laboratory, in a: collaborator's group, and in the field of proteomics allow a new and direct approach to this largely unexplored, frontier in the study of plasticity. A novel procedure for inducing bursts of activity in the Drosophila nervous system is shown to initiate plasticity processes that lead to gene expression changes associated with-establishment of long-term plasticity. A technique called Differential Gel Electrophoresis (DIGE) has the ability to visualize and compare levels of about 5000 different protein elements from Drosophila heads. Combining these two techniques with mass spectrometry and more established molecular genetic approaches, the proposed experiments will identify proteins whose activities are rapidly regulated by post-translational covalent modification, by regulated protein turnover, or by induction of translation, following appropriate neural activity. Drosophila melanogaster is an excellent model organism for these analyses. The commonality of underlying mechanisms involved in plasticity regulation in mammals and insects is indicated by the functional conservation of almost all known regulators of plasticity in both phyla. However, the rate of progress of the proposed analyses in Drosophila is much faster, facilitated not only by its short generation time and facility for genetics, but also by novel resources from Drosophila genome projects, proteomic technologies and newly developed procedures for gene disruption, perturbation and replacement in vivo. The work is significant because it addresses a particularly poorly studied area, of fundamental, importance in synaptic remodeling events that underlie behavioral change. By identifying rapidly regulated, neuronal proteins the program may contribute new, early molecular markers of plasticity processes that underlie addiction. In addition, results from these experiments may identify new molecules to targets for pharmacological therapy.

描述：（申请人提供） 依赖经验的神经可塑性是吸毒成瘾的基础，复杂， 持久的行为改变；吸毒后的结果。分子迅速 受神经活动调节可能非常重要 可塑性过程。因此，他们的识别和特征是 与理解基础的细胞机制的特别相关 行为改变。该建议旨在将新颖方法结合起来 确定经过快速，转录后调节的分子 建立长期突触变化。最近的技术进步 首席调查员的实验室，A：合作者小组和 蛋白质组学领域允许采用新的直接方法来实现这一尚未探索的， 塑性研究的边界。诱导爆发的新型程序 果蝇神经系统中的活性显示出启动可塑性 导致基因表达变化的过程与建立有关 长期可塑性。一种称为差异凝胶电泳的技术 （DIGE）具有可视化和比较约5000种不同水平的能力 果蝇头的蛋白质元素。 将这两种技术与质谱和更建立的技术相结合 分子遗传方法，提出的实验将鉴定蛋白质 其活动受到翻译后共价迅速调节 通过调节的蛋白质更新或通过诱导翻译来修饰， 遵循适当的神经活动。果蝇Melanogaster是一个很好的 这些分析的模型生物体。基本机制的共同点 参与哺乳动物和昆虫的可塑性调节是由 两者中几乎所有已知可塑性调节剂的功能保护 门。但是，果蝇中提出的分析的进度率是 更快的速度，不仅可以促进其短期和设施 遗传学，以及果蝇基因组项目的新资源， 蛋白质组学技术和新开发的基因破坏程序， 体内扰动和替代。 这项工作很重要，因为它解决了一个特别糟糕的研究 基本的重要性，在突触重塑事件中的重要性 行为改变。通过识别快速调节的神经元蛋白 程序可能会贡献新的，早期的可塑性过程标记 那是成瘾的基础。此外，这些实验的结果可能 将新分子确定为药理治疗靶标。