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Analysis of Genes Induced by Bone Morphogenetic Protein (BMP) in the Early Stage of Response

Research Project

Project/Area Number 06807156
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Surgical dentistry
Research InstitutionTokyo Medical and Dental University

Principal Investigator

ASAHINA Izumi  Tokyo Medical and Dental University Faculty of Dentistry, Instructor, 歯学部, 助手 (30221039)

Co-Investigator(Kenkyū-buntansha) OIDA Shinichiro  Tokyo Medical and Dental University Faculty of Dentistry, Instructor, 歯学部, 助手 (10114745)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsBMP / OP-1 / bone induction / gene / 骨形成因子 / 骨芽細胞 / 分化
Research Abstract

To reveal the mechanism of action of Bone Morphogenetic Protein (BMP), we have examined the genes induced by BMP in the early stage of the response, which may control the action of BMP,using the differential display method in this study.
We used Osteogenic Protein-1 (OP-1, BMP-7) as BMP,because highly purified recombinant OP-1 was available from Creative BioMolecule Inc. Examination of the effect of OP-1 on several cell lines revealed that OP-1 enhances alkaline phosphatase activity, one of the maker of osteoblastic characteristics, in the culture of osteoblastic MC3T3-E1 cells from mouse calvaria and stromal ST-2 cells from mouse bone marrow. After the treatment of OP-1 to these cells for 24hours, total RNA was extracted with AGPC method and reverse transcription was performed. According to the instruction of the kit for differential display method, polymerase chain reaction (PCR) on the resulted cDNA was performed with 20 sets of primer combination under the presence of 35S-dATP.The amplified cDNAs were electophoretically separated on a 6% sequence gel. Analysis of bands on the auto radiogram revealed that 66 cDNA fragments from MC3T3-E1 and 11 cDNA fragments from ST-2 were expressed differentially with or without the treatment of OP-1. cDNAs from MC3T3-E1 were subcloned and sequences were analyzed. There were a few clones which have homology with ubiquitin, SP-1, etc.but most of them were not identified clearly because the acquired sequences were only 200 to 300 base in 3'end region. Northern analysis was performed using these cDNA clones as a probe, but expression of most of them did not have any difference between with and without the treatment of OP-1. Therefore, we assumed that many cDNA clones obtained by differential display methods were psudepositive. However, we got two cDNA clones which show apparent difference on Northern analysis from ST-2. Sequencing analysis is on procedure.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] J. C. Maliakal. I. Asahina, et al.: "Osteogenic Protein-1 (BMP-7) Inhibits Cell Proliferation and Stimulate the Expression of Markers characteristics of Osteoldaste" Growth Factors. 11. 227-234 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Y. Maruoka, I. Asahina, et al.: "Production of Functional Human Bone Morphogenetic Protein-2 Using a Bconlovirns/Sf-9 Insect Cell System" Biochem Mol. Biol. Int.35. 957-963 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] I. Asahina, K. Sampath. et al: "Hunan Osteogenic Protein-1 Induces Chordwldastic Osteoblastic and / or Adripocytic Differatiatia" Exp. Cell. Res.222. 38-47 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] J.C.Maliakal, I.Asahina, P.V.Hauschka, T.K.Sampath: "Osteogenic Protein-1 (BMP-7) Inhibits Cell Proliferation and Stimulates the Expression of Markers Characteristic of Osteoblast Phenotype in Rat Osteosarcoma (17/2.8) Cells" Growth Factors. Vol 11. 227-234 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Y.Maruoka, S.Oida, T.Iimura, K.Takeda, I.Asahina, S.Enomoto, and S.Sasaki: "Production of Functional Human Bone Morphogenetic Protein-2 Using a Baculovirus/ SF-9 Insect Cell System" Biochem Mol.Biol.Int.Vol 35 (5). 657-963 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] I.Asahina, T.K.Sampath, P.V.Hauschka: "Human Osteogenic Protein-1 (OP-1) Induces Chondroblastic, Osteoblastic and/or Adipocytic Differentiation of Clonal Murine Target Cells" Exp.Cell Res.vol.222. 38-47 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Iimura, T., Oida, S., Takeda, K., Maruoka, Y.and Sasaki, S: "Changes in Homeobox-containing gene expression during ectopic bone formation induced by bone morphogenetic protein" Biochem.Biophys.Res.Commun. 201. 980-987 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Takeda, K., Oida, S., Goseki, M., Iimura, T., Maruoka, Y.Amagasa, M., and Sasaki, S: "Expression of bone morphogenetic protein gene in the human dental pulp cells" Bone. 15. 467-470 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Oida, S., Iimura, T., Maruoka, Y., Takeda, K.and Sasaki, S: "Cloning and sequence of bone morphogenetic protein-4 (BMP-4) from a human placental cDNA library" DNA Sequence. 5. 273-275 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Y.Maruoka,et al.: "Production of Functional Human Bone Morphogenetic Protein-2 Using a Bacnlovims/Sf-9 Insect Cell System" Biochem.Mol.Biol.Int.35. 957-963 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] I.Asahina,et al.: "Human Osteogenic Protein-1 lnduces Chondroblastic,Ostesbiastic,and/or Adipocytic Differentiation of Clonrl Murine Target Cells" Exp.Cell Res.222. 38-47 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] S.Oida,et al.: "Cloning and sequence of bone morphogenetic protein-4(BMP-4)from a human placental cDNA library" DNA sequence. 5. 273-275 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Y.Maruoka.I.Asahina et.al: "Production of Functional Human Bone Morphogenentic Protein-2 Using A Baculovirus/SF-9 Cell System." Biochem. Mol.Biol.Int.in press. (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] J.C Maliakal.I.Asahina et al.: "Osteogenic Protein-1(BMP-7) Fnhibits Cell Prolifevation and Stimulates the Expression of Markers Characteristics of Osteoblast Phemotype" Grouth Factors. 11. 227-234 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Takeda,S.Oida et al: "Molecular Cloning of rat bone inorphogenetic Protein(BMP)Type IA receptor and its expression during ectopic bone formation" Biochem Biophys.Res.Commun.204. 203-209 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Iimura,S.Oida et al: "Modulation of responses to TGF-β by 1,25-Dihydvoxy v.tamin D_3 in MG-63 osteoblastic cells" Biochen.Biophys.Res.Commur.204. 918-923 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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