| Project/Area Number |
61580158
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Tokyo Metropolitan Institute of Gerontology |
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Principal Investigator |
KAWASHIMA Seiichi Department of Biochemistr,Tokyo Metropolitan Institute of Gerontology, 生化学部, 室長 (60008571)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Masami Department of Biochemistry,Tokyo Metropolitan Institute of Gerontology, 生化学部, 助手 (00192279)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1987: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Calcium-activated neutral protease / Monoclonal antibody / 免疫組織化学 / カルシウム依存性 / プロテアーゼ |
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| Research Abstract |
To determine the physiological role of calcium-activated neutral protease (CANP), monoclonal antibodies against CANP were prepared and used for localization of CANP in tissues.Two kinds of CANP with different Ca^<2+>-sensitivities were purified from rabbit skeletal muscle and mice were immunized with them.Hybridoma cells of immunized spleen and myeloma cells were screened by ELISA and cloned.Fifteen and 77 clones were obtained which secreted antibodies against mCANP and uCANP,respectively.Most of them recognized the large subunit of either mCANP or uCANP but one antibody bound to the large subunits of both mCANP and uCANP and another one bound only to the small subunit of both CANPs.Immunohistochemical staining of frozen sections of rabbit organs by ABC method showed that epithelial tissues reacted intensely.These tissues incluce great alveolar and squamous alveolar cells in 1ung;interlobular artery,vein,and bile duct in liver;small vessels in skeletal muscle;glomeruli,jixtanglomerular cells,distal and collecting tubules in kindney;mucous epithelium in gallbladder;interstitial cells in testis; and cuboical epithelial cells in brain choroid plexus.On the other hand,hepatocytes,epithelial cells which have ill defined basal lamina,were stained very faintly.These observations suggest that the physiological function of CANP is involved with transport systems in epithelial tissues through basal lamina.
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