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Histochemical and functional aspects of glycolipids

糖脂的组织化学和功能方面

基本信息

批准号：
13670024
负责人：
KAWAKAMI Hayato
金额：
$ 2.18万
依托单位：
Kyorin University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

Histochemical localization of various kinds of glycolipids was investigated with the use of confocal laser scanning microscopy and transmission electron microscopy. Rat various organs prefixed with 4% formaldehyde and 1% glutaraldehyde were cut into nonfrozen slices (40μm thick) and treated with monoclonal antibodies against glycolipids (anti-GM1, GD2, GD3, GT1b, GQ1b, Gb3 etc.) followed by treatment with Cy3-labeled socondary antibody for light microscopy or peroxidase-labeled socondary antibody for electron microscopy. Some of the specimens from neuronal organs were also stained with specific antibodies against neurotransmitters (GABA, glutamate etc.) by Cy5 labeling method. In cerebellar cortex, each ganglioside revealed specific distribution pattern, e.g. anti-GQ1b antibody was positive for the plasma membrane of Purkinje cell body and its dendrites, Golgi cisternae, part of the nerve endings around the Purkinje cell body. These areas especially the nerve endings around the Purkinje cell body were also positive for anti-GABA antibody. In small intestine, Auerbach's plexus and outer muscular layer were specifically stained for some ganglioside antibodies e.g. GM1, GD2. Basolateral plasma membrane of intestinal gland epithelial cells were specifically stained for anti-Gb3. Specific distribution of glycolipids in various organs seem to have some specific relationships with neuronal functions and other cellular functions.

应用激光共聚焦扫描显微镜和透射电子显微镜对各种糖脂的组织化学定位进行了研究。用4%甲醛和1%戊二醛预固定的大鼠各器官切成40μm厚的非冷冻切片，用抗糖脂单克隆抗体（抗GM 1、GD 2、GD 3、GT 1b、GQ 1b、Gb 3等）处理。随后用Cy 3标记的soconucleotide抗体处理用于光学显微镜检查或用过氧化物酶标记的soconucleotide抗体处理用于电子显微镜检查。部分神经器官标本也用抗神经递质（GABA、谷氨酸等）的特异性抗体染色。Cy 5标记法。在小脑皮质，每种神经节苷脂显示出特异性分布模式，如抗GQ 1b抗体对浦肯野细胞体及其树突的质膜、高尔基池、浦肯野细胞体周围的部分神经末梢呈阳性。这些区域尤其是浦肯野细胞体周围的神经末梢也呈GABA抗体阳性。在小肠，奥尔巴赫氏神经丛和外肌层对某些神经节苷脂抗体如GM 1、GD 2有特异性染色。肠腺上皮细胞的基底外侧质膜被特异性地染色用于抗-Gb 3。糖脂在各器官中的特异性分布似乎与神经元功能和其他细胞功能有某些特定的关系。