| Project/Area Number |
07044235
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Niigata University |
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Principal Investigator |
SHIMIZU Fujio Institure of Nephrology, Niigata University School of Medicine professor, 医学部, 教授 (40012728)
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| Co-Investigator(Kenkyū-buntansha) |
KAWACHI Hiroshi Institute of Nephrology, Niigata University School of Medicine Research Assistan, 医学部, 助手 (60242400)
MORIOKA Tetsuo Institute of Nephrology, Niigata University School of Medicine Research Assistan, 医学部, 助手 (00210146)
SALANT David j. Rend section, Boston University Medical Center professor, 医学部, 教授
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| Project Period (FY) |
1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Monoclona antibody / Proteinuria / Slit diaphragm / Podocyte / Glomerular permeability / Functional molecule / Cultured glomerular epithelial cell |
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| Research Abstract |
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.
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