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Functional molecules on glomerular epithelial cells

肾小球上皮细胞的功能分子

基本信息

批准号：
07044235
负责人：
SHIMIZU Fujio
金额：
$ 1.41万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至无数据
项目状态：
已结题

项目摘要

Cultured glomerular epithelial cells in the loboratory of Dr. Salant in Boston University could not be stained by the monoclonal antibody (mAb) 5-1-6.Cloning of the target antigenic molecule has not been succeeded, using the rat kidney cDNA library prepared in the Boston laboratory.The traial to get the antigenic molecule has been repeated in vain, using the affinity column with in vitro prepared- and purified-mAb 5-1-6 and the solublized rat glomeruli.The biogenesis of the target antigen of mAb 5-1-6 was studied in the developing glomerulus by immunolocalization and metabolic labeling. This antigen first became faintly, but clearly, detectable on the basal and lateral sides of the developing podocytes at the S-shaped body stage. Staining intensity increased with further maturation and was restricted to the visceral epithelial cells. On immunoelectro microscopy, the antigen was seen along the basal and lateral surfaces below occluding junctions at the early capillary loop stage and later, with the interdigitation of foot processes, became concentrated in the slit pores. At no stage was the antigen seen on the apical surface. Metaboric labeling studies showed that the antigenic molecule is actively synthesized during initial glomerular development and that the rate of synthesis declines substantially with maturation.From these results cDNA-library was considered to be more poperly prepared from rat glomeruli in neonatal stage for cloning of this antigenic molecule. We are planning to clone this, applying the cDNA library from baby glomeruli.

波士顿大学Salant博士实验室培养的肾小球上皮细胞不能被单克隆抗体（mAb） 5-1-6染色。使用波士顿实验室制备的大鼠肾脏cDNA文库克隆目标抗原分子尚未成功。利用体外制备和纯化的mab 5-1-6和溶解的大鼠肾小球的亲和柱进行了多次获得抗原分子的试验，但没有成功。通过免疫定位和代谢标记研究了mAb 5-1-6靶抗原在发育中的肾小球的生物发生。在s形体阶段，这种抗原首先在发育中的足细胞的基侧和外侧变得微弱但清晰。染色强度随着进一步成熟而增加，并且仅限于内脏上皮细胞。在免疫电镜下，抗原在早期毛细血管袢阶段沿着闭塞连接处的基面和侧面可见，后来随着足突的交叉，抗原集中在狭缝孔中。未见抗原出现在根尖表面。代谢标记研究表明，抗原分子在初始肾小球发育期间积极合成，随着成熟，合成速度大幅下降。由此可见，从新生期大鼠肾小球中制备cdna文库是克隆该抗原分子的较好选择。我们正计划克隆它，利用来自婴儿肾小球的cDNA文库。