Modulation of AMPA-Type Receptor Trafficking in Aplysia

描述(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。皮尔是加州大学洛杉矶分校生理科学系和神经生物学系的教授。在他的大部分科学生涯中，他一直在研究无脊椎动物生物体学习和记忆的简单形式的神经基础。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受到了高度培训。最近来自PI实验室的数据表明，内源性单胺5-羟色胺(5-HT)调节海螺运动神经元AMPA型谷氨酸受体的细胞内转运。来自pl实验室的其他数据表明，5-羟色胺诱导的AMPA型受体运输的调节有助于海兔的行为敏化。为了阐明5-羟色胺诱导海兔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方面的研究技能。具体地说，在获奖期间，PI将进行以下研究项目：(I)从海兔神经系统克隆和测序离子型谷氨酸受体基因；(Ii)进行原位杂交，以定位克隆的谷氨酸受体在海兔中枢神经系统的mRNAs；(Iii)在非洲爪哇卵母细胞中表达海兔谷氨酸受体，用于对所表达的受体通道进行生物物理和药理学表征；(Iv)产生不同谷氨酸受体亚单位的抗体，用于免疫组织化学研究5-羟色胺对受体分布的影响；以及iv)测试海兔运动神经元在长期记忆、长期敏感化过程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名同事合作。建议的合作者在完成建议的研究所需的分子和细胞生物技术方面都有很高的知识和经验。此外，合作者的实验室拥有拟议研究所需的设备。

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.