NEUROBIOLOGY OF SULFATED GLUCURONYL GLYCOCONJUGATES

硫酸化葡萄糖醛酰糖复合物的神经生物学

• 批准号: 2265208
• 负责人: FIROZE B JUNGALWALA
• 金额: $ 21.87万
• 依托单位: EUNICE KENNEDY SHRIVER CTR MTL RETARDATN
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1995-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2265208
• 关键词: beta galactosidase; beta glucuronidase; cell adhesion; cell cell interaction; cell differentiation; cell migration; cerebellar cortex; cerebral cortex; developmental neurobiology; electron microscopy; enzyme mechanism; extracellular matrix; galactosyltransferases; glycolipids; glycoprotein biosynthesis; glycoproteins; high performance liquid chromatography; immunocytochemistry; laboratory mouse; laboratory rabbit; laboratory rat; lipid metabolism; microscopy; neuronal guidance; protein purification; proteoglycan; sulfatases; sulfotransferase; tissue /cell culture; beta galactosidase; beta glucuronidase; cell adhesion; cell cell interaction; cell differentiation; cell migration; cerebellar cortex; cerebral cortex; developmental neurobiology; electron microscopy; enzyme mechanism; extracellular matrix; galactosyltransferases; glycolipids; glycoprotein biosynthesis; glycoproteins; high performance liquid chromatography; immunocytochemistry; laboratory mouse; laboratory rabbit; laboratory rat; lipid metabolism; microscopy; neuronal guidance; protein purification; proteoglycan; sulfatases; sulfotransferase; tissue /cell culture

The major long term goal of this project is to understand the role and function of sulfoglucuronyl glycolipids (SGGLs) and other neolacto series of glycolipids which are stage specific and developmentally regulated antigens in the cortex in the central nervous system, during the important period of cell differentiation and cell migration. The carbohydrate moiety of SGGLs is highly immunogenic and monoclonal antibodies such as HNK-1, which bind to the carbohydrate also react with either the same or similar epitope on several important glycoproteins involved in cellular interaction, such as neural cell adhesion molecules, N_CAMs, L1, J1, tenascin, cytotactin, integrin, NILE, myelin associated glycoprotein and PO protein. The proper development of the nervous system depends upon these molecules. Although SGGLs are expressed in the cortex during embryonic and neonatal development only, they are associated with Purkinje cells and in peripheral nervous system, even in the adult. The present specific aims are to understand the biochemical regulation of their differential expression in these areas of the nervous system by studying the enzymes involved in the metabolism of these lipids. Enzymes include several glycosyltransferases and glycosidases. Besides SGGLs other neolactoseries of glycolipids having the same backbone, such as fuconeolactotetraoside and ganglioside LD1 are also developmentally regulated antigens to their proper location in the cortex during development. Cells expressing specific carbohydrates will be studied in vitro to evaluate the role of these antigens in maturation and differentiation. SGGLs and sulfatide act as ligand cell-adhesion between cell-cell, cell-substratum and bind extracellular matrix protein laminin. Studies are planned to characterize a 30 kD SGGL/sulfatide binding protein from developing cortex and cerebellum. The protein and antibodies to this protein will be used as probes to investigate the role of SGGLs and of the HNK-1 reactive proteins in cellular interactions. New fixation method has been developed to differentiate immunocytochemical localization of HNK-1 reactive lipids versus proteins in neural tissues. This method will be used to precisely localize the expression of lipid versus protein antigens, at light and electron microscopic level during development of the cortex and cerebellum, and in certain mutants. Cellular and subcellular localization of these molecules may provide new clues as to their functional role.

该项目的主要长期目标是了解角色和 磺核糖糖（SGGLS）和其他Neolacto系列的功能 特定于阶段且受发展调节的糖脂 中枢神经系统皮质中的抗原，重要 细胞分化和细胞迁移的时期。 碳水化合物部分 SGGL的高度免疫原性和单克隆抗体，例如HNK-1， 与碳水化合物结合的也与相同或相似的反应 几种参与细胞的重要糖蛋白的表位 相互作用，例如神经细胞粘附分子，N_CAM，L1，J1， 替氏蛋白，细胞动物，整合素，尼罗河，髓磷脂相关糖蛋白和PO 蛋白质。 神经系统的正确发展取决于这些 分子。 尽管在胚胎期间，SGGL在皮质中表示 仅新生儿发育，它们与Purkinje细胞和 即使在成年人中，周围神经系统也是如此。 目前的特定目标 要了解其差异的生化调节 通过研究酶在神经系统的这些区域中的表达 参与这些脂质的代谢。 酶包括几种 糖基转移酶和糖苷酶。 除了SGGLS其他新分子店 具有相同主链的糖脂，例如fuconeolactotetraoside和 神经节苷脂LD1也对其适当的开发调节抗原 开发过程中皮质的位置。 表达特异性的细胞 将在体外研究碳水化合物，以评估这些作用 抗原成熟和分化。 SGGL和亚磺胺为 细胞细胞，细胞肌间和结合之间的配体细胞粘附 细胞外基质蛋白层粘连蛋白。 计划的研究表征 从发育的皮质和 小脑。 该蛋白质的蛋白质和抗体将被用作 研究SGGL和HNK-1反应性蛋白的作用的探针 在细胞相互作用中。 已开发出新的固定方法 分化HNK-1反应性脂质的免疫细胞化学定位 与蛋白质相对于神经组织中的蛋白质。 此方法将用于精确 将脂质与蛋白质抗原的表达定位在光和 电子显微镜水平在皮质和小脑发育过程中， 在某些突变体中。 这些细胞和亚细胞定位 分子可以提供有关其功能作用的新线索。