| Project/Area Number |
01570018
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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 |
General anatomy (including Histology/Embryology)
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| Research Institution | Keio University, School of Medicine, Department of Anatomy |
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Principal Investigator |
YAMASHITA Shuji Keio Univ., Sch. of Med., Assistant Professor, 医学部, 講師 (90050666)
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| Co-Investigator(Kenkyū-buntansha) |
SHOZAWA Masahide Keio Univ., Sch. of Med., Lecturer, 医学部, 助手 (50170840)
AISO Sadakazu Keio Univ., Sch. of Med., Assistant Professor, 医学部, 講師 (60138013)
YASUDA Kenjiro Keio Univ., Sch. of Med., Professor, 医学部, 教授 (90050327)
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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 |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Common antigens of secretory granule membrane / Monoclonal antibody / Immunohistochemistry / Recycling of secretory granule membrane / Immunoelectron microscopy / Rat / Rabbit / 分泌果粒共通抗原 |
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| Research Abstract |
Monoclonal antibodies (mAbs) were generated to the secretory granule membrane of rat and rabbit parotid gland, respectively. The mAbs recognized integral membrane glycoproteins localized on the luminal side of the secretory granules with N-linked carbohydrates : molecular weight ; 98-65 kD in the rat, 55-65 kD in the rabbit.
Immunohistochemical studies indicated that both the secretory granule membrane antigens were found in the secretory granules of both exocrine and endocrine cells, and in the lysosomes of various cells in the rat and rabbit. These results indicated that common antigens are present in the secretory granule membranes. Immunoelectron microscopy with immunogold revealed that antigen was present on the membrane of the secretory granules, lysosomes, the Golgi vesicles and condensing vacuoles in pancreatic and parotid acinar cells and AR-42J rat pancreatic tumor cells ; the Golgi stacks exhibited no immunoreaction. In the rabbit, the antigen was localized exclusively along the membranes of secretory granules in the parotid acinar cells, exocrine pancreatic cells and adenohypophyseal cells.
Employing HRP labeled mAb, we followed the retrieval of the antigen after exocytosis in AR42J cells. The mAb was internalized specifically with antigen-mediated endocytosis. It was transported to endosomes, subsequently to the trans-Golgi network, and then packaged into secretory granules. However, the Golgi stacks revealed no uptake of the labeled antibody. These results indicated that the common antigen of secretory granule memrane is reused after exocytosis, and suggested that the pool of the antigen might present in the trans-Golgi network.
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