ASSEMBLY OF A NEURAL PLASMA MEMBRANE PROTEOLIPID

这笔赠款的重点是质膜的组装。 突触质膜蛋白脂蛋白(PMPLP)和 在髓鞘中。该项目将利用一系列细胞生物学 从其合成引用中跟踪该蛋白质的技术 神经元和少突胶质细胞连接到各自的靶膜， 例如。突触质膜和髓鞘。决心会 膜组装过程中PMPLP的加工 专注于蛋白质的酰化。的相对增长率 这种蛋白质组装成髓鞘的过程将被确定，并 与其他髓鞘蛋白相比，如主要的髓鞘 蛋白脂和髓鞘相关糖蛋白。涂覆的 囊泡将从白质中分离出来，并存在 PMPLP和其他髓鞘蛋白的测定和研究将 以确定这些蛋白质是否是新的 合成的。我们将使用免疫学方法尝试选择 以及可视化涂层囊泡的亚群 运输PMPLP或其他髓鞘蛋白。决赛 PMPLP在大鼠髓鞘组装膜中的定位 脊髓将使用冷冻-超微切开术和 免疫电子显微镜。研究将与反 PMPLP和胶体金共轭秒数的可视化 抗体。质膜蛋白脂在体内的组装 将对突触质膜进行研究。使用免疫学 我们将检查技术是否，在灰质衍生涂层 水泡，PMPLP定义了水泡的一个特定子类，并且 是否能将子类与携带 突触小泡蛋白。PMPLP本土化到专门化 突触的膜结构域将使用冷冻- 超薄冰冻切片的免疫电子显微镜观察 大鼠齿状回。PMPLP在突触的存在将是 在不同年龄的动物身上进行分析，当一个标记 突触发生和成熟的突触在 大脑区域。突触功能障碍与精神障碍的关系 发育迟缓和癫痫表明这种主要的突触蛋白 可能同时参与生理学和病理生理学。 大脑。此外，它还与髓鞘有关 提示它不仅可能与髓鞘的完整性有关 但这种蛋白质组装的异常可能会导致 上升到与星座脑功能障碍相关的 脱髓鞘和髓鞘过少疾病。

The focus of this grant is the assembly of the plasma membrane proteolipid protein (PMPLP) in the synaptic plasma membrane and in myelin. This project will utilize a series of cell biologic techniques to follow this protein from its cite of synthesis in neurons and oligodendrocytes to the respective target membranes, eg. synaptic plasma membrane and myelin. Determination will be made of processing of PMPLP during the membrane assembly focusing on the acylation of the protein. The relative rate of assembly of this protein into myelin will be determined and compared to other myelin proteins such as the major myelin proteolipid and the myelin associated glycoprotein. Coated vesicles will be isolated from white matter and the presence of PMPLP and other myelin proteins determined and studies will be carried out to determine if these proteins have been newly synthesized. We will use immunologic methods to try to select as well as visualize subpopulations of coated vesicles that may transport the PMPLP or other myelin proteins. The final localization of PMPLP in the assembled myelin membrane of rat spinal cord will be examined using cryo- ultramicrotomy and immunoelectron microscopy. Studies will be carried out with anti PMPLP and visualized with colloidal gold conjugated second antibody. The assembly of the plasma membrane proteolipid into synaptic plasma membranes will be studied. Using immunological techniques we will examine if, in grey matter derived coated vesicles, PMPLP defines a specific sub class of vesicles and whether the sub class can be distinguished from those carrying synaptic vesicle proteins. The localization of PMPLP to specific membrane domains at the synapse will be determined using cryo- immunoelectron microscopy on ultra thin frozen sections of the rat dentate gyrus. The presence of PMPLP at the synapse will be analyzed in animals at different ages when a marked synaptogenesis and maturation of synapses is taking place in this brain region. The association of synaptic dysfunction with mental retardation and epilepsy suggests that this major synaptic protein could be involved in both the physiology and pathophysiology of the brain. Moreover, its associated with the myelin sheath suggest that not only may it be involved in the integrity of myelin but the abnormalities in the assembly of this protein could give rise to a constellation brain dysfunction associated with demyelinating and hypomyelinating diseases.