Modulation of AMPA-Type Receptor Trafficking in Aplysia

海兔 AMPA 型受体运输的调节

• 批准号: 7000303
• 负责人: DAVID L GLANZMAN
• 金额: $ 11.76万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-06 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7000303
• 关键词: AMPA receptors; Aplysia; Xenopus oocyte; behavioral habituation /sensitization; confocal scanning microscopy; gene expression; immunocytochemistry; in situ hybridization; intracellular transport; long term memory; molecular cloning; motor neurons; receptor expression; sensorimotor system; serotonin; synapses

DESCRIPTION (provided by applicant): The purpose of this application is to provide the PI with resources that will enable him to acquire technical skills and expertise in molecular and cell biology. The PI is a Professor in the Departments of Physiological Science and Neurobiology at UCLA. During most of his scientific career the PI has studied the neural basis of simple forms of learning and memory in invertebrate organisms. The PI is highly trained in electrophysiological techniques for investigating learning-related neuronal plasticity, as well as in behavioral techniques for investigating simple forms of invertebrate learning and memory. Recent data from the laboratory of the PI indicates that the endogenous monoamine, serotonin (5-HT), modulates the intracellular trafficking of AMPA-type glutamate receptors in motor neurons of the marine snail Aplysia californica. Additional data from the Pl's laboratory suggests that 5-HT-induced modulation of AMPA-type receptor trafficking contributes to behavioral sensitization in Aplysia. To elucidate the potential cellular mechanisms of 5-HT-induced modulation of AMPA-type receptor trafficking in Aplysia, the PI wishes to acquire research skills in cellular and molecular biology. Specifically, during the award period the PI will undertake the following research projects: (i) to clone and sequence ionotropic glutamate receptor genes from the Aplysia nervous system; (ii) to perform in situ hybridization to localize the mRNAs of the cloned glutamate receptors in the Aplysia CNS; (iii) to express the Aplysia glutamate receptors in Xenopus oocytes for biophysical and pharmacological characterization of the expressed receptor channels; (iv) to generate antibodies to the different glutamate receptor subunits for use in immunohistochemical studies of the effect of 5-HT on the distribution of the receptors; and iv) to test whether expression of the glutamate receptor is enhanced in motor neurons of Aplysia during a form of long-term memory, long-term sensitization. To develop the technical expertise required for these projects, the PI will collaborate with several UCLA colleagues. The proposed collaborators are all highly knowledgeable and experienced in the molecular and cell biological techniques necessary for completing the proposed research. Furthermore, the laboratories of the collaborators have the necessary equipment for the proposed research.

描述（由申请人提供）：本申请的目的是为PI提供资源，使其能够获得分子和细胞生物学方面的技术技能和专业知识。PI是加州大学洛杉矶分校生理科学和神经生物学系的教授。在他的大部分科学生涯中，PI研究了无脊椎生物简单形式的学习和记忆的神经基础。PI在研究学习相关神经元可塑性的电生理技术以及研究无脊椎动物学习和记忆的简单形式的行为技术方面受过高度训练。PI实验室的最新数据表明，内源性单胺，5-羟色胺（5-HT），调节运动神经元中的AMPA型谷氨酸受体的细胞内运输的海洋蜗牛Aparasia californica。来自PI实验室的额外数据表明，5-HT诱导的AMPA型受体运输的调节有助于失智症中的行为敏化。为了阐明5-HT诱导的AMPA型受体转运调节的潜在细胞机制，PI希望获得细胞和分子生物学方面的研究技能。具体而言，在授予期间，主要研究者将开展以下研究项目：（i）克隆和测序来自失智症神经系统的离子型谷氨酸受体基因;（ii）进行原位杂交，以定位失智症CNS中克隆的谷氨酸受体的mRNA;（三）在非洲爪蟾卵母细胞中表达谷氨酸受体，用于表达的受体通道的生物物理学和药理学表征;（iv）产生针对不同谷氨酸受体亚单位的抗体，用于5-HT对受体分布的影响的免疫组织化学研究;和（iv）测试在一种形式的长期记忆、长期致敏过程中，谷氨酸受体的表达是否在失智症的运动神经元中增强。为了发展这些项目所需的技术专长，PI将与加州大学洛杉矶分校的几位同事合作。建议的合作者都是在完成拟议的研究所需的分子和细胞生物学技术方面知识渊博和经验丰富。此外，合作者的实验室拥有拟议研究所需的设备。