FUNCTION OF GENES AND PROTEINS IN LEARNING AND MEMORY

基因和蛋白质在学习和记忆中的功能

• 批准号: 2891771
• 负责人: ARNOLD none ESKIN
• 金额: $ 18.5万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2891771
• 关键词: Aplysia; RNase protection assay; alternatives to animals in research; conditioning; gel electrophoresis; genetic transcription; genetic translation; immunocytochemistry; immunologic assay /test; long term memory; molecular cloning; molecular psychobiology; nerve /myelin protein; neural information processing; neural plasticity; neurogenetics; nucleic acid sequence; protein purification; protein sequence; reflex; serotonin; western blottings

The major objective of this research is to provide insights into the regulation of biochemical and molecular events that occur while organisms learn and form memories. We are examining the hypothesis that regulation of gene transcription and protein translation is involved in the formation of long-term memories. Moreover, a cascade of transcription events is hypothesized to affect the translation of a diverse array of proteins which control a broad spectrum of cellular functions. The specific framework of our studies is sensitization of the defensive tail-siphon withdrawal reflex in Aplysia, and mRNA and protein changes in large homogeneous clusters of bilaterally-symmetrical sensory neurons in the pleural ganglia. These sensory neurons are key components of the reflex and its plasticity. This proposal has four Specific Aims which are designed to extend our previous research in this area. Specific Aim 1 is to discover proteins whose synthesis is regulated by treatments that mimic some effects of sensitization training and then characterize their temporal responses. Specific Aim 2 is to obtain partial amino acid sequences and gene sequences of proteins or mRNAs that are affected by induction treatments. Specific Aim 3 is to determine if behavioral training or 5-HT alters levels of specific mRNAs in sensory neurons and define the time domains in which changes occur. Specific Aim 4 is to obtain antibodies to an Aplysia Tolloid-like protein that is similar to a previously-identified, developmentally-regulated protein. We will use the antibodies to localize the protein in neural tissue and to examine the relationship between changes in Tolloid mRNA and changes in the synthesis of the Tolloid protein. The research will use quantitative analysis of proteins separated by two dimensional gel electrophoresis, peptide sequencing, gene cloning and sequencing, and other techniques of molecular biology. Ribonuclease protection assays will be used to measure levels of mRNAs. Antibodies will be used in several types of studies including Western blotting and immunocytochemistry. This research will provide substantial new insights into the mechanisms for the induction, storage and read-out of a form of long-term memory. Moreover, the research will provide general information on the role of specific proteins and genes in cellular function and plasticity.

这项研究的主要目的是提供有关 生物体发生时发生的生化和分子事件的调节 学习并形成记忆。我们正在研究以下假设：监管 基因转录和蛋白质翻译的参与参与形成 的长期记忆。 此外，一系列转录事件是 假设影响多种蛋白质的翻译 它控制广泛的细胞功能。具体的 我们研究的框架是防御性尾虹吸管的敏化 海兔的退缩反射以及 mRNA 和蛋白质的巨大变化 两侧对称感觉神经元的同质簇 胸膜神经节。这些感觉神经元是反射的关键组成部分 及其可塑性。 该提案有四个具体目标，分别是 旨在扩展我们之前在该领域的研究。 具体目标 1 是 发现其合成受模拟治疗调节的蛋白质 敏化训练的一些效果，然后描述其特征 时间反应。具体目标2是获得部分氨基酸 受以下因素影响的蛋白质或 mRNA 的序列和基因序列 诱导治疗。 具体目标 3 是确定行为是否 训练或 5-HT 会改变感觉神经元中特定 mRNA 的水平 定义发生变化的时域。具体目标 4 是 获得与海兔 Tolloid 样蛋白类似的抗体 先前鉴定的发育调节蛋白质。 我们将使用 抗体定位神经组织中的蛋白质并检查 Tolloid mRNA 变化与合成变化之间的关系 Tolloid 蛋白。 该研究将使用定量分析 二维凝胶电泳分离蛋白质、肽 测序、基因克隆和测序以及其他分子生物学技术 生物学。核糖核酸酶保护测定将用于测量 mRNA。 抗体将用于多种类型的研究，包括 蛋白质印迹和免疫细胞化学。 这项研究将提供 对诱导、储存机制的实质性新见解 并读出某种形式的长期记忆。 此外，该研究将 提供有关特定蛋白质和基因的作用的一般信息 细胞功能和可塑性。