| Project/Area Number |
05454176
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Human pathology
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| Research Institution | Tokai University School of Medicine |
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Principal Investigator |
MORIUCHI Tetsuya Tokai University School of Medicine Associate Professor Department of Molecular Life Science, 医学部, 助教授 (20174394)
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| Co-Investigator(Kenkyū-buntansha) |
TANIGUCHI Yasushi Tokai University School of Medicine Assistant Professor Department of Molecular, 医学部, 講師 (30207188)
YOSHIMURA Shinichi Tokai University School of Medicine Assistant Professor Department of Molecular, 医学部, 講師 (30230808)
TSUTSUMI Yutaka Tokai University School of Medicine Associate Professor Department of Pathology, 医学部, 助教授 (80138643)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1994: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1993: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | Homeobox protein / Homeobox gene / Immunohistochemistry / Cell adhesion / Integrin / Fibronectin / 免疫組織化学 / ホメオボックス / ホメオドメイン / 抗体 / プログラム細胞死 / ヒト / マウス |
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| Research Abstract |
Polyclonal antibody against amino acid sequence encoded by first exon of HOXD4 gene was prerapred in rabbits. Distribution of HOXD4 protein was investigated in day-11, -13, -15, and -17 mouse embryos by using the HOXD4 antibody. (1) Skin ; Upper 1/3 of skin layrs was stained by anti-HOXD4. Lower 3/4 skin layrs was stained by anti-PCNA (marker of proliferating cells). Inter-digital skin was stained with anti-HOXD4 but not with anti-PCNA.(2) Cartilage ; Ossification center of the cartilage but not immature cartilage-forming cells were stained by anti-HOXD4. (3) Kidney ; Medullary region was stained by anti-HOXD4.
HOXD3 gene was inserted at sense or antisense orientation in expression vector, and the construct was transfected in human erythroleukemia HEL cells. Two cell clones, E1 and E6, which were overexpressing HOXD3, were obtained. E1 and E6 expressed 7-10 fold amounts of HOXD3 mRNA compared to parental HEL cells, and C1 and C2 cell clones which had been, trasfected with antisense HOXD3 gene. Cell-cell and cell-extracellular matrix adhesions were markedly enhanced in E1 and E2 cells. Growth rate did not differ among sense-transfec-tants, parental HEL cells and antisense-transfectants. Overexpression of HOXD3 increased the mRNA level of integrin beta3. FACS analysis revealed that expression of alpha IIbbeta3 (GPIIb-IIIa) complex (=fibronectin receptor) on the cell surface was strongly increased in HEL sense-transfectants, E1 and E6.
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