MYELIN MEMBRANE STRUCTURE: STABILITY & PATHOLOGY

总体目的：表征脂质的分子结构 以及神经髓鞘膜中的蛋白质;并确定 稳定膜阵列的相互作用。 的相关性 生物物理和生物化学技术（包括X射线衍射， 电镜，SDS聚丙烯酰胺凝胶电泳， 免疫印迹和薄层色谱法）应用于不同类型 样本（包括整个未固定或固定的组织，组织匀浆， 组织组分，以及脂质和蛋白质的模型系统）将用于 回答以下具体问题 (1)层间致密物的精细结构和组成是什么 中枢神经系统髓鞘连接处 计算机图像分析将在 电子显微照片。 将使用SDS-PAGE/免疫印迹法鉴别 组织组分中的蛋白质在连接处富集。 x射线 对比增强后，将记录完整CNS髓鞘的模式 以及在连接处富集的组织部分。 (2)髓鞘中的脂质和蛋白质是怎样排列的？ 相关性 将对选定的神经元进行生化和X射线测量 小鼠的逻辑突变体，某些遗传学上更古老的脊椎动物， 周围神经系统内的不同神经， 髓鞘制剂。 髓鞘结构的改变 将分析特定的金属阳离子。 结构和双分子层间 由非变性蛋白脂重构多层膜的相互作用 蛋白质（PLP）加髓鞘脂质，PLP加髓鞘碱性蛋白将是 分析了 (3)脱髓鞘的结构基础是什么？ 的结构和 钙离子载体和补体对中枢神经系统髓鞘的生化作用将 下定决心。 层间紧密连接的完整性将是 在实验性过敏动物的白色物质中检查 脑脊髓炎和尸检材料的人与多个 硬化症

Overall objective: To characterize the molecular organization of lipids and proteins in nerve myelin membranes; and to define the molecular interaction that stabilize the membrane arrays. A correlation of biophysical and biochemical technique (including X-ray diffraction, electron microscopy, SDS polyacrylamide gel electrophoresis, immunoblotting, and thin-layer chromatography) applied to different types of specimens (including whole unfixed or fixed tissue, tissue homogenates, tissue fractions, and model systems of lipids and proteins) will be used to address the following specific questions (1) What is the fine-structure and composition of the interlamellar tight junctions of CNS myelin? Computer image analysis will be carried out on electron micrographs. SDS-PAGE/immunoblotting will be used to identify proteins in tissue fractions that are enriched in the junctions. X-ray patterns will be recorded from intact CNS myelin after contrast enhancement of the junctions, and also from tissue fractions enriched in the junctions. (2) What is the arrangement of lipids and proteins in myelin? Correlation between biochemical and X-ray measurements will be made for selected neuron logical mutants of the mouse, certain phylogenetically-older vertebrates, different nerves from within the peripheral nervous system, and isolated myelin preparations. Structural modifications of myelin induced by specific metal cations will be analyzed. The structure and inter-bilayer interaction of multilayers reconstituted from non-denatured proteolipids protein (PLP) plus myelin lipids, and PLP plus myelin basic protein will be analyzed. (3) What is the structural basis of demyelination? The structural and biochemical effects of calcium-ionophore and complement on CNS myelin will be determined. The integrity of interlamellar tight junctions will be examined in white matter from animals having experimental allergic encephalomyelitis and from autopsy material of humans with multiple sclerosis.