Modulation of AMPA-Type Receptor Trafficking in Aplysia

描述（由申请人提供）：本申请的目的是为PI提供资源，使他能够获得分子和细胞生物学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系和神经生物系的教授。在他的大部分科学生涯中，PI研究了无脊椎生物中简单形式的学习和记忆的神经基础。PI在研究与学习相关的神经元可塑性的电生理技术以及研究简单形式的无脊椎动物学习和记忆的行为技术方面训练有素。来自PI实验室的最新数据表明，内源性单胺- 5-羟色胺（5-HT）调节加州海蜗牛运动神经元中ampa型谷氨酸受体的细胞内运输。来自Pl实验室的其他数据表明，5- ht诱导的ampa型受体运输调节有助于澳大利亚的行为致敏。为了阐明5- ht诱导的ampa型受体转运的潜在细胞机制，PI希望获得细胞和分子生物学的研究技能。具体而言，在资助期内，该计划将进行以下研究项目：(i)克隆和测序海百合神经系统的嗜离子性谷氨酸受体基因；（ii）原位杂交定位克隆的谷氨酸受体mrna；（iii）在爪蟾卵母细胞中表达谷氨酸受体，对表达的受体通道进行生物物理和药理学表征；（iv）产生针对不同谷氨酸受体亚基的抗体，用于5-HT对受体分布影响的免疫组织化学研究；iv)测试在长时记忆、长时致敏过程中，运动神经元中谷氨酸受体的表达是否增强。为了开发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。拟议的合作者在完成拟议的研究所需的分子和细胞生物学技术方面都有很高的知识和经验。此外，合作者的实验室有必要的设备进行拟议的研究。

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.