SORTING AND TRANSPORT OF SPECIFIC NEURONAL GLYCOPROTEINS

特定神经元糖蛋白的分选和运输

• 批准号: 3404190
• 负责人: RICHARD T AMBRON
• 金额: $ 15.21万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3404190
• 关键词: Golgi apparatus; autoradiography; cell adhesion; chromatography; electrophoresis; histochemistry /cytochemistry; hybridomas; immunochemistry; immunoelectron microscopy; interneurons; monoclonal antibody; neuronal transport; neurons; neurotransmitters; oligosaccharides; saltwater environment; synapses

The work in this proposal addresses two issues of fundamental importance to our understanding of neuronal function and development. The first is to determine how membrane glycoproteins synthesized in the cell body of a neuron are selectively routed to the pre- and postsynaptic terminal. These processes are central to the proper assembly of surface membranes and to how distinct regions of the neuronal plasma membrane are maintained in a functionally and structurally differentiated state. The second concerns the possible role of glycoproteins as mediators of interneuronal recognition and adhesion. My colleagues and I intend to explore these points using a single cell - R2 the giant neuron of Aplysia californica. R2's cell body, presynaptic, and postsynaptic regions are anatomically accessible. Moreover, R2 synthesizes only ten membrane glycoproteins, a remarkable simplicity relative to the complexity of glycoprotein fractions from vertebrate nervous tissue. Two of R2's glycoproteins have distinctly different destinations in the cell: glycoprotein-I is the major glycoprotein on the cell surface of the cell soma whereas glycoprotein-V is preferentially exported into the axon where it is rapidly transported towards R2's synapses. The site at which the two glycoproteins are sorted will be determined by immunocytochemistry at the ultrastructural level using monoclonal antibodies raised to each glycoprotein. In addition, the oligosaccharide chains of both glycoproteins will be characterized with the idea that a modification of a glycosyl moiety is the signal responsible for the sorting. R2, GCN, and R15 are three large identified neurons that differ in function, location, and neurotransmitter type. We will examine the glycoproteins present on the surface of the cell body and terminals of these neurons in order to test directly the hypothesis that neurons can be distinguished by their glycoprotein constituents. Many features of the proposed experiments are unique and present a unified, multidisciplinary approach to issues of major importance to our knowledge of the neuron.

该提案中的工作解决了两个基本重要性的问题 我们对神经元功能和发展的理解。 首先是 确定膜糖蛋白如何在A的细胞体中合成 神经元有选择地路由到突触前和突触后末端。 这些 过程对于正确组装表面膜和到达至关重要 神经元质膜的不同区域如何保持在A中 在功能和结构上分化的状态。 第二个问题 糖蛋白作为神经元介质的可能作用 识别和粘附。 我的同事和我打算探索这些 使用单个细胞-R2点的点为加州Aplysia的巨型神经元。 R2的细胞体，突触前和突触后区域在解剖学上是 可访问。 此外，R2仅合成十个膜糖蛋白，一个 相对于糖蛋白分数的复杂性，显着的简单性 来自脊椎动物神经组织。 R2的两个糖蛋白在该糖蛋白中具有明显不同的目的地 细胞：糖蛋白-i是细胞表面上的主要糖蛋白 细胞体soma，而糖蛋白-V优先导出到轴突中 它迅速运输到R2的突触。 该网站 两种糖蛋白分类将通过免疫细胞化学确定 在超微结构级别，使用每种抗体的单克隆抗体 糖蛋白。 另外，两者的寡糖链 糖蛋白将以一种修改的想法来表征 糖基部分是负责分类的信号。 R2，GCN和R15是三个大型鉴定的神经元，不同 功能，位置和神经递质类型。 我们将检查 存在于细胞体和末端的糖蛋白 这些神经元为了直接检验神经元可以是 由其糖蛋白成分区别。 许多功能 提议的实验是独特的，并提出了统一的多学科 解决对我们对神经元知识的重要性问题的方法。