MHC CLASS I EXPRESSION AND FUNCTION IN MURINE CEREBELLUM

小鼠小脑中 MHC I 类的表达和功能

• 批准号: 6574833
• 负责人: Kevin D Phelan
• 金额: $ 7.1万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6574833
• 关键词: behavior test; biological signal transduction; cerebellar Purkinje cell; cerebellum; developmental neurobiology; electrophysiology; gene targeting; genetically modified animals; immunocytochemistry; in situ hybridization; laboratory mouse; major histocompatibility complex; neural plasticity; protein structure function; synapses; synaptogenesis; western blottings

DESCRIPTION (provided by applicant): Major histocompatibility complex class I (MHC-I) molecules are well known for the important immune role that they perform in the presentation of viral and tumor antigens to circulating T lymphocytes. However, they also play a critical "non-immune" role in activity-dependent synapse elimination during development and in synaptic plasticity in the adult. Differences in various classical and non-classical MHC-I expression patterns suggests that specific MHC-I proteins may play regionally specific roles in the CNS. However, our knowledge of MHC-I function in the CNS is limited such that we do not know if classical and/or non-classical MHC-I proteins are selectively required for synapse elimination and/or synaptic plasticity. This application will take advantage of the unique synaptic circuitry of the cerebellum to compare and contrast the potential roles of classical and non-classical MHC-I molecules during normal cerebellar development and function at the single cell level. The specific aims of this study are designed to: 1) test the hypothesis that developmental changes in specific MHC-I expression patterns in the cerebellar cortex can be correlated with known patterns of synaptogenesis in the murine cerebellum; and 2) determine if blockade of classical or nonclassical MHC-I expression or signaling will result in impaired synapse elimination at the climbing fiber- Purkinje cell synapse and/or altered synaptic plasticity at the parallel fiber-Purkinje cell synapse in the adult cerebellum. These aims will be achieved using standard immunohistochemistry and in situ hybridization techniques (using MHC-I subtype specific antibodies and mRNA probes) in combination with electrophysiological recording of cerebellar Purkinje cells in cerebellar slices prepared from adult wildtype and selected knockout mice. The results of the proposed experiments will extend our limited knowledge of neuronal MHC-I expression and function within the CNS to include the cerebellum, an important structure involved in both motor control and cognition. More importantly, this study will begin to provide essential information on the specific non-immune roles that various classical and non-classical MHC-I proteins may play in the developing and adult CNS.

描述（由申请人提供）：主要的组织相容性复合物I类（MHC-I）分子以其在病毒和肿瘤抗原表现为循环T淋巴细胞中所扮演的重要免疫作用而闻名。 但是，它们在发育过程中的活动依赖性突触消除和成人突触可塑性中也起着关键的“非免疫”作用。 各种经典和非经典MHC-I表达模式的差异表明，特定的MHC-I蛋白可能在中枢神经系统中起特定于区域的作用。 但是，我们对中枢神经系统中MHC-I功能的了解受到限制，因此我们不知道经典和/或非古典MHC-I蛋白是否有选择地消除突触消除和/或突触可塑性。 该应用将利用小脑的独特突触电路比较和对比在正常小脑发育和单个细胞水平上的正常小脑发育和功能期间经典和非经典MHC-1分子的潜在作用。 这项研究的具体目的旨在：1）检验以下假设：小脑皮层中特定MHC-I表达模式的发育变化可以与鼠小脑中的突触发生模式相关联； 2）确定在攀岩纤维 - 珀kinje细胞突触中的封锁或非经典MHC-I表达或信号传导是否会导致突触消除受损和/或在成人小脑中平行纤维纤维螺旋藻细胞突触的突触可塑性改变。 这些目标将使用标准的免疫组织化学和原位杂交技术（使用MHC-I亚型特异性抗体和mRNA探针）结合使用成人野生野生型和选定的基因敲除小鼠制备的小脑伯氏切片中小脑Purkinje细胞的电生理记录。 提出的实验的结果将扩展我们对中枢神经系统内神经元MHC-I表达和功能的有限了解，以包括小脑，这是运动控制和认知涉及的重要结构。 更重要的是，这项研究将开始提供有关各种经典和非古典MHC-I蛋白可能在发展中和成人中枢神经系统中起作用的特定非免疫作用的基本信息。