SULFOGLUCURONYL (HNK-1) CARBOHYDRATE IN NEUROGENESIS

本研究的长期目标是验证硫磺脲基碳水化合物（SGC）及其结合蛋白（SBP）的相互作用涉及神经元上的SBP-1及其配体SGC对神经元和胶质细胞过程的作用，从而提供一种指导（SGC）及其结合蛋白（SBP）参与神经元上的SBP-1及其配体SGC对神经元和胶质细胞过程的作用，从而为细胞迁移和过程生长提供一种指导和信号机制。在这四个目标中计划了大量的实验来实现这一目标。具体目的1是研究胚胎大脑皮层外植体培养和厚层组织培养系统中神经突的生长和神经元迁移。将添加各种抗体和糖脂来确定对迁移和神经生成的影响。具体目标2是克隆GlcNAc-Tr，该酶被认为是SGGC前体Lc0se3Cer合成的关键调节酶。主要的方法将是表达克隆，替代使用他们最近构建的抑制减法杂交cDNA文库。具体目的3是研究GT基因表达在发育过程中对糖脂和糖蛋白的SGC合成的调控作用。一种假设是，GlcNAc- tr将参与硫糖醛基糖脂（SGGL）的表达，而不是携带SGC表位的糖蛋白，这可能会利用其他GlcNAc转移酶。

The long term goal of this study is to test the hypothesis that interaction of sulfoglucuronyl carbohydrate (SGC) and its binding protein (SBP) involve SBP-1 on neurons and its ligand SGC on processes on neurons and glia, thus providing a guidance (SGC) and its binding protein (SBP) involve SBP-1 on neurons and its ligand SGC on processes of neurons and glia, thus providing a guidance and signaling mechanism for cell migration and process outgrowth. A large number of experiments toward that goal are planned in the four aims. Specific Aim 1 is to study neurite outgrowth and neuronal migration in explant cultures of embryonic cerebral cortex and thick slice tissue culture systems. Various antibodies and glycolipids will be added to determine the effect on migration and neuritogenesis. Specific Aim 2 is to clone the GlcNAc-Tr that has been described as the key regulatory enzyme for synthesis of Lc0se3Cer, the SGGC precursor. The main approach will be expression cloning, with the alternative use of a suppression subtractive hybridization cDNA library they recently constructed. Specific Aim 3 is to investigate the role of GT gene expression in regulation of SGC synthesis on glycolipids and glycoproteins during development . One hypothesis is that GlcNAc-Tr will be involved in expression of sulfoglucuronyl glycolipid (SGGL) rather than glycoproteins bearing the SGC epitope, which would presumably utilize other GlcNAc transferases.