Immunocytochemical study on the behavior and function of osteoclasis using monoclonal antibodies to osteoclasts
| Project/Area Number |
03670896
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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 |
Conservative dentistry
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| Research Institution | Kyusyu University |
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Principal Investigator |
AKAMINE Akifumi Kyusyu University, Faculty of Dentistry, Associate Professor, 歯学部, 助教授 (00117053)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Ryusei Kyusyu University, Faculty of Dentistry, Assitant, 歯学部, 助手 (20205008)
AIDA Yoshitomi Kyusyu University, Faculty of Dentistry, Lecture, 歯学部, 講師 (10127954)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1992: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1991: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | bone resorption / osteoclast / osteoblast / immunohistochemistry / sialoglycoprotein / monoclonal antibody / osteopontin / リソゾーム性膜シアロ糖タンパク質 / モノクロ-ナル抗体 |
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| Research Abstract |
Osteoclasts are primary cells responsible for bone resorption. However, the precise mechanism as to osteoclastic bone resorption remains unclear. We therefore examined immunocytochemically the behavior and the function of osteoclasts and osteoblasts in bone rmodeling.
1.Immunocytochemical localization of a major lysosomal membrane sialoglycoprotein (LGP 107) in osteoclasts : The immunocytochemical localization was investigated of LGP 107 using rat osteoclasts at various stages of differentiation. LGP 107 was exclusively confined to the apical plasma mambrane at the ruffled border of the active osteoclasts. The protein was also concentrated in a number of endocytic vacuoles near the ruffled border membrane. However, the post and/or resting osteoclasts were totally devoid of the membraneous localization of LGP 107. These results indicate that the protein is largely synthesized in the active osteoclast and rapidly translocated to the ruffled border membrane. LGP 107 is suggested to contrib
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ute to the formation and maintenance of the specialized acidic environment for bone reorption.
2.Expression and localization of LGP 107 in osteoblastic lineage cells : The immunocytochemical localization of LGP 107 was investigated in osteoblast linage cells involved in osteoclastic bone resorption. Strong immunoreaction products for LGP 107 occurred on the plasma membranes in the osteoblasts and osteocytes prior to the appearance of osteoclasts. Furthermore, strong reactions were also observed on the plasma membranes in the osteoblastic cells adjacent to the active osteoclasts. These data suggest that LGP 107 in osteoblastic cells and osteocytes may play an important role in cell-recognition and/or cell-adhesion, and that LGP 107 may be involved in osteoblastic degradation of the osteoid as well as exposure of the bone surface.
3.Production of monoclonal antibodies to osteoclasts by in vitro immunization : The characteristics of a monoclonal antibody produced against produced against osteoclast-like multinucleated cells (MNC_S) formed in rat bone marrow cultures were examined immunohistochemically and biochemically. After the in vitro immunization was performed, the monoclonal antibody HOK1 was obtained. This antibody reacted weakly with stromal cells and intensely with both MNC_S and their putative migratory traces on culture dishes. Western blotting using purified rat osteopontin verified that the antigen recognized by HOK1 was osteopontin. Postive HOK1 immunoreactivity was further observed in the resorption lacunae formed by a culture of MNC_S. The present data suggested that osteopontin is preferentially present on the resorption lacunae in resorbing calcified matrices and that osteoclasts might trap this protein on their cell surface. Less
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Report
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Research Products
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