ASSEMBLY OF A NEURAL PLASMA MEMBRANE PROTEOLIPID

这项资助的重点是质膜的组装 突触质膜中的蛋白脂质蛋白（PMPLP）， 髓磷脂 该项目将利用一系列细胞生物学 技术跟踪这种蛋白质从其合成的网站， 神经元和少突胶质细胞到各自的靶膜， eg. 突触质膜和髓磷脂。 决心将 在膜组装过程中， 专注于蛋白质的酰化。 的相对速率 将确定该蛋白质组装成髓磷脂， 与其他髓磷脂蛋白如主要髓磷脂相比， 蛋白脂质和髓磷脂相关糖蛋白。 涂覆 囊泡将从白色物质中分离出来， PMPLP和其他髓鞘蛋白的测定和研究将在 以确定这些蛋白质是否是新的 合成了 我们将使用免疫学方法来尝试选择， 以及可视化包被囊泡的亚群， 运输PMPLP或其他髓鞘蛋白。 最终 PMPLP在大鼠组装髓鞘膜上的定位 将使用冷冻超微切片术检查脊髓， 免疫电镜 研究将与反 PMPLP，并用胶体金偶联的第二 抗体的 质膜蛋白脂质组装成 将研究突触质膜。 使用免疫学 技术，我们将研究，如果在灰质衍生涂层 PMPLP定义了囊泡的特定子类， 是否可以将子类与携带 突触囊泡蛋白。 PMPLP定位于特定的 突触处的膜结构域将使用冷冻- 超薄冰冻切片免疫电镜观察 大鼠齿状回。 突触处PMPLP的存在将是 在不同年龄的动物中进行了分析， 突触的发生和成熟都发生在 大脑区域。 突触功能障碍与精神分裂症的关系 发育迟缓和癫痫表明这种主要的突触蛋白 可能涉及到的生理和病理生理 大脑 此外，它与髓鞘有关， 这表明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 会导致一系列脑功能障碍， 脱髓鞘和髓鞘生成不足疾病。

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.