Functional molecules on glomerular epithelial cells

点击翻译按钮获取中文摘要

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.