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Cytochemical studies of the intracellular processing of glycoconjugates

糖复合物细胞内加工的细胞化学研究

基本信息

批准号：
02670015
负责人：
MURATA Fusayoshi
金额：
$ 1.15万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

1)The analysis of glycosylation using postembedding staining method.Cis side of the Golgi apparatus was labeled with DBA, HPA and SBA, trans side and secretory granules were stained with HPA, PNA and RCA-I, UEA-I and LFA labeled transmost Golgi apparatus. By this method, the process of glycosylation of the Golgi apparatus was clearly analyzed.2)The analysis of glycosylation using preembedding staining method.MPA labeled perinuclear space, inner lumen of rough endoplasmic reticulum and cis side of the Golgi apparatus. DBA stained the same region of the Golgi apparatus. PNA labeled the intermediate and RCA-1, UEA-1 and LFA stained the trans side of the Golgi apparatus. Secretory granules were labeled with RCA-1, UEA-1 and LFA. The results-obtained with these two methods were not inconsistent.3)The analysis of glycosylation process using monoclonal antibodies to ABO substances.Anti A labeled the whole lamellae of the Golgi apparatus, while anti B and anti H labeled the trans side of the Golgi apparatus. The difference of the labeling was noticed among antibodies.4)The development of a new cytochemical method to stain acidic glycoconjugates.By making cationic colloidal gold with poly-L-lysine, changing pH of the staining solution and setting many control experiments, we established a new staining method of acidic glycoconjugates. This method is used both in light microscopic level and electron microscopic level. The sites of sialylation and sulfation were analyzed using this method and found that both localized at the trans side of the Golgi apparatus.5)The improvement of cytochemical demonstration method of sulfated glycoconjugates.By adding physical development step to high iron diaminethiocarbohydrazide-silver proteinate method, we succeeded in improving the staining sensitivity of this method. This method allows us to develop in the bright field condition. The site of sulfation has been analyzed and found to localize at the trans Golgi apparatus.

1)包埋后染色分析：用DBA、HPA、SBA标记高尔基体顺侧，用HPA、PNA、RCA-I、UEA-I、LFA标记高尔基体的反面和分泌颗粒。用该方法对高尔基体的糖基化过程进行了清晰的分析。2)用包埋前染色方法分析了高尔基体的糖基化过程。DBA对高尔基体的同一区域进行染色。PNA标记中间体，RCA-1、UEA-1和LFA标记高尔基体的反面。用RCA-1、UEA-1和LFA标记分泌颗粒。3)用抗ABO物质的单抗对糖基化过程进行分析，抗A标记高尔基体的全层，抗B和抗H标记高尔基体的反面。4)建立了酸性糖偶联物的细胞化学染色新方法，通过用多聚L赖氨酸制备阳离子胶体金，改变染色溶液的pH值，并设置多个对照实验，建立了一种新的酸性糖结合物的染色方法。这种方法既适用于光学显微镜水平，也适用于电子显微镜水平。5)硫酸糖偶联物细胞化学显示方法的改进，通过在高铁二氨基硫代氨基脲-银蛋白法的基础上加入物理显影步骤，成功地提高了该方法的染色灵敏度。这种方法使我们可以在明亮的视场条件下展开。硫化的位置已经被分析并发现定位在反式高尔基体上。