| Project/Area Number |
01540594
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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 |
動物発生・生理学
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| Research Institution | Osaka University (1990)
Nagoya University (1989) |
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Principal Investigator |
NAKANISHI Yasuo College of Bio-Medical Technology Professor, 医療技術短期大学部, 教授 (40022636)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Submandibular Epithelium / Morphogenesis / Epithelial-mesenchymal Interaction / Collagen / Cleft Formation / Heparitinase / Extracellular Matrix / Heparin Sulfate Proteoglycan / ヘパリチナ-ゼ / 唾液腺 / 上皮-間充織相互作用 / プロテオヘパラン硫酸 / モノクロ-ン抗体 / マウス |
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| Research Abstract |
The major purpose of this study was to identify extracellular matrix components which could regulate the cleft formation in the epithelium of the mouse embryonic submandibular gland. The results obtained are as follows :
1. The physiological function of heparin sulfate chains during morphogenesis was studied by the use of heparitinase purified from Flavobacterium heparinum. Addition of the enzyme in the culture medium halted the branch-in tiation and changed the round epithelial shape to elongated one, together with concomitant reduction in lobular growth. This effect was mimicked by the addition of heparin sulfate and heparin in the culture medium. However, no effect was observed with chondroitin sulfate, showing that heparin sulfate chains in the rudiment play a crucial role (s) in the branching morphogenesis.
2. Intact, non-degraded organ rudiments from 13-day fetus were successfully used as antigens to raise rat monoclonal antibodies against mouse embryonic submandibular gland, lung and kidney, and more than 60 different hybridomas were obtained. The first group of hybridomas had specificities toward fibrillar structures or dotted structures in the extracellular matrix of submandibular mesen-chyme. The second group of hybridomas was found to be reactive to basement membranes of various tissues. The third group was that they reacted only to the basement membrane areas closely associated with the stabilized epithelial structures including stalk and clefts. The fourth group specifically stained various types of epithelia of lung and kidney. The findings strongly suggest that the method of monoclonal antibody production through the immunization of a rat with intact organ rudiments are valuable in identifying novel components in the extracellular matrix and further in evaluating their roles in the branching morphogenesis of submandibular gland, lung and kidney.
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