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Microscale cDNA subtraction method for detection of regulators for maxillo-facial development.

Research Project

Project/Area Number 08557095
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Morphological basic dentistry
Research InstitutionOsaka University

Principal Investigator

KURISU Kojiro  Osaka University Faculty of Dentistry, Professor, 歯学部, 教授 (50028346)

Co-Investigator(Kenkyū-buntansha) SATO Katsuhiko  Chugai Pharmaceutical Co., LTD., 研究者
TABATA Makoto  Osaka University, Faculty of Dentistry, Assistant, 歯学部, 助手 (20243248)
KATO Joji  Osaka University, Faculty of Dentistry, Assistant, 歯学部, 助手 (90243245)
IWAMOTO Masahiro  Osaka University, Faculty of Dentistry, Instructor, 歯学部, 講師 (30223431)
WAKISAKA Satoshi  Osaka University, Faculty of Dentistry, Associate Professor, 歯学部, 助教授 (40158598)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥10,300,000 (Direct Cost: ¥10,300,000)
Fiscal Year 1997: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1996: ¥8,800,000 (Direct Cost: ¥8,800,000)
KeywordsOrofacial tissue / Gene / Gene multiplication / Rabbit / Morphogenesis / Development / Tissue differentiation / cDNA library / 遺伝子増幅 / ウサギ / cDNAライブラリー
Research Abstract

Subtractive hybridization methods are used to detect genes that are expressed in a specific tissue or cell but not in the other. Usually cDNA called "tester" that contains differentially expressed genes is hybridized with an excess amount of RNA called "driver" that does not contain such genes. As a result, differentially expressed cDNA in enriched in unhybridized cDNA fraction. Although traditional subtractive hybridization methods have been successfully used in some cases, they require a lot of tester and driver mRNA.Therefore, it has been difficult to identify differentially expressed genes when only small amount of tissue (e.g.mouse tooth bud etc.) is available as a starting material.
In the present study, we constructed chondrocytes specific subtraction cDNA library by two different methods : [1] conventional single strand subtraction (tester cDNA-driver mRNA) and [2] double strand cDNA subtraction (tester dscDNA-driver dscDNA). In both methods, we used PCR for amplifying tester cDNA.We found the quality of those libraries was similar but the amount of tester mRNA that was required for library construction was much smaller in the method [2] than that of [1]. In both libraries, most of cDNA clones encodes 3' untranslated sequence plus partial sequence and the average size of inserted cDNAs was about 550bp. We speculated this was due to the limitation of PCR amplification. Although partial cDNA may be useful as a probe to examine expression pattern of the gene in the tissue, It is necessary screen the unsubtracted library to obtain full length cDNA.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (61 results)

All Other

All Publications (61 results)

  • [Publications] S.H.Youn: "Growth-associated protein-43 (GAP 43) in the regenerating periodontal Ruffini endings of the rat incisor following injury of the inferior alveolar nerve." Brain Res.(in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.G.Liu: "Developmental role of PTHrP in the murine molars." Europ.J.Oral SCi.106. (S)143-146 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Miyawaki: "Calbindin D28k-like immuno-reactivity in the developing and degenerating circumvallate papilla of the rat." Cell Tssue Res.291. 81-90 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Enomoto-Iwamoto: "Bone morphogenetic protein signaling is required for maintenance of differentiating activity, cessation of proliferation and terminal differentiation of chondrocytes." J.Cell Biol.40. 409-418 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Kurisu: "Human genes for dental anomalies." Oral Dis.3. 223-228 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Inai: "Immunohistochemical localization of connexin 43 in the enamel organ of the rat upper incisor during ameloblast development." Arch.Histol.Cytol.60. 297-306 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] N.Kinto: "Fibroblasts expressing sonic hedgehog induce osteoblast differentiation and proliferation of chondrocytes." FEBS Lett.404. 319-323 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Nakamura: "Induction of osteogenic differentiation by hedgehog proteins." Biochem.Biophys.Res.Commun.237. 465-469 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Enomoto-Iwamoto: "Involvement of α5β1 integrin in matrix interactions and proliferation of chondrocytes." J.Bone Miner.Res.in press. (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Miyawaki: "Calbindin D28k-like immunoreactive nerve fibers in the predentin of rat molar teeth." Archs.oral Biol.42. 773-777 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.H.Youn: "Regeneration of periodontal primary afferents of the rat incisor following injuru of the inferior alveolar nerve with special reference to neuropeptide Y-like immunoreactive primary afferents." Brain Res.752. 161-169 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.J.Tabata: "Hepatocyte growth factor is involved in the morphogenesis of tooth germ in murine molars." Development. 122. 1243-1251 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Wakisaka: "Partial coexistence of neuropeptide Y and calbindin D38K in the trigeminal ganglion following peripheral axotomy og the inferior nerves of the rat." Brain Res.707. 228-234 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Wakisaka: "Distribution and possible origin of galanin-like immunoreactive nerve fibers on the mammalian dental pulps." Regul.Peptides. 62. 137-143 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Wakisaka: "Protein product 9.5 in the developing mouse circumvallate papilla : comparision with neuron specific enolase and calcitonin gene related peptide." Anat.Embryol.194. 365-372 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.J.Tabata: "Expression of cytokeratin 14 in ameloblast-lineage cells of the developing tooth of rat both in vivo and in vitro." Archs.oral Biol.41. 1019-1027 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Kawakami: "BMP signaling during bone pattern determination in the developing limb." Development. 122. 3557-3567 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Aikawa: "Establishment of bone morphogenetis protein 2 respons 2 responsive chondrogenic cell line." J Bone Miner.Res.11. 544-553 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakisaka, S., et al.: "Partial coexistence of neuropeptide Y and calbindin D38K in the trigeminal ganglion following peripheral axotomy of the inferior alveolar nerve of the rat." Brain Res.707. 228-234 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tabata, M.J., et al.: "Hepatocyte growth factor is involved in the morphogenesis of tooth germ in murine molars." Development. 122. 1243-1251 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato, J., et al.: "Immunohistochemical changes in the distribution of nerve fibers in the periodontal ligament during an experimental tooth movement of the rat molar." Acta Anat.157. 53-62 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakisaka, S., et al.: "Distribution and possible origin of galanin-like immunoreactive nerve fiber on the mammalian dental pulps." Regul.Pepties. 62. 137-143 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakisaka, S., et al.: "Protein product 9.5 in the developing mouse cirumvallte papilla ; comparison wirh neuron specific enolase and calcitonin gene related peptide." Anat.Embryol.194. 365-372 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakisaka, S., et al.: "Neuropeptide Y-like immunoreactive primary afferents in the dental pulp and periodontal ligament following peripheral nerve injury to the inferior alveolar nerve in the rat." Brain Res.712. 11-18 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tabata, M.J.et al.: "Expression of cytokeratin 14 in ameloblast-lineage cells of the developing tooth of rat both in vivo and in vitro." Archs.oral Biol. 41. 1019-1027 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kawakami, Y.et al.: "BMP signaling during bone pattern determination in the developing limb." Development. 122. 3557-3566 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Aikawa, T.et al.: "Establishment of bone morphogenic protein 2 responsive chondrogenic cell line." J.Bone Miner.Res.11. 544-553 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Matsuo, S.et al.: "Influense of fluoride in secretory pathway of the secretory ameloblast in rat incisor tooth germs exposed to sodium fluoride." Arch.Toxicol.70. 420-429 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Youn, S.H.et al.: "Regeneration of periodontal primary afferents of the rat incisor following injury of the inferior alveolar nerve with special reference to neuropeptide Y-like immunoreactive primary afferents." Brain Res.752. 161-169 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Miyawaki, Y.et al.: "Calbindin D28k-like immunoreactive nerve fibers in the predentin of rat molarteeth." Archs.oral Biol.42. 773-777 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Enomoto-Iwamoto, M.et al.: "Involvement of alpha5beta1 integrin in matrix interactions and proliferation of chondrocytes." J.Bone Miner.Res.12. 1124-1132 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nakamura, T.et al.: "Induction of osteogenic differentiation by hedgehog proteins." Biochem.Biophys.Res.Commun.237. 465-469 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kinto, N.et al.: "Fibroblasts expressing sonic hedgehog induce osteoblast differentiation and proliferation of chondrocyte." FEBS Lett.404. 319-323

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Inai, T.et al.: "Immunohistochemical localization of connexin 43 in the enamel organ of the rat upper incisor during ameloblast development." Arch.Histol.Cytol.60. 297-306 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kurisu, K.et al.: "Human genes for dental anomalies." Oral Dis.3. 223-228 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Enomoto-Iwamoto, M.et al.: "Bone morphogenetic protein signaling is required for maintenance of differentiating activity, cessation of proliferation and terminal differentiation of chondrocytes." J.Cell.Biol.40. 409-418 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Liu, J.G.ret al.: "Developmental role of PTHrP in the murine molars." Europ.J.Oral Sci.106 (Suppl.1). 143-146 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Miyawaki, Y.et al.: "Calbindin D28k-like immunoreactivity in the developing and degenerating circumvallate papilla of the rat." Cell Tssue Res.291. 81-90 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Youn, S.H.et al.: "Growth-associated protein-43 (GAP 43) in the regenerating periodontal Ruffini endings of the rat incisor following injury of the inferior alveolar nerve." Brain Res.(in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.G.Liu: "Dvelopmental role of PTHrP in the murine molars." Europ.J.Oral Sci.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.H.Youn: "Growth-associated protein-43 (GAP 43) in the regenerating periodontal Ruffini endings of the rat incisor following injury of the inferior alveolar nerve." Brain Res.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Miyawaki: "Calbindin D28K-like immuno-reactivity in the developing and degenerating circumvallate papilla of the rat." Cell Tssue Res.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Kurisu: "Human genes for dental anomalies." Oral Dis.3. 223-228 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Inai: "Immunohistochemical localization of connexin 43 in the enamel organ of the rat upper incisor during ameloblast development." Arch.Histol.Cytol.60. 297-306 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Nakamura: "Induction of osteogenic differentiation by hedgehog proteins." Biochem.Biophys.Res.Commun.237. 465-469 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] M.Enomoto-Iwamoto: "Bone morphogenetic protein signaling is required for maintenance of differentiating activity,cessation of proliferation and terminal differentiation of chondrocytes." J.Cell Biol.40. 409-418 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] N.Kinto: "Fibroblasts expressing sonic hedgehog induce osteoblast differentiation and proliferation of chondrocytes." Febs lett.404. 319-323 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] M.Enomoto-Iwamoto: "Involvement of α5β1 integrin in matrix interactions and proliferation of chondrocytes." J.Bone Miner.Res.(in press). (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Miyawaki: "Calbindin D28k-like immunoreactive nerve fibers in the predentin of rat molar teeth." Archs.oral Biol.42. 773-777 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.H.Youn: "Regeneration of periodontal primry afferents of the rat incisor following injuru of the inferior alveolar nerve with special reference to neuropeptide Y-like immunoreactive primary afferents." Brain Res.752. 161-169 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Matsuo: "Influense of fluoride in secretory pathway of the secretory ameloblast in rat incisor tooth germs exposed to sodium fluoride." Arch.Toxicol.70. 420-429 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Wakisaka: "The effects of peripheral nerve injury of the masseteric nerve on the levels of calcium binding proteins and neuropeptide Y,and their correlation in the mesencephalic trigeminal nucleus of the rat." Brain Res.735. 249-256 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Aikawa: "Establishemnt of bone morphogenic protein 2 responsive chondrogenic cell line." J.Bone Miner.Res.11. 544-553 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Miyawaki: "Calbindin D28k-like immunoreactivity in the gustatory epithelium in the rat." Neurosci.Lett.214. 29-32 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Kawakami: "BMP signaling during bone pattern determination in the developing limb." Development. 122. 3557-3566 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 栗栖浩二郎: "アンチセンスオリゴDNAによる細胞機能の解析" ブレインサイエンス. 7. 81-89 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.J.Tabata: "Hepatocyte growth factor is involved in the morphogenesis of tooth germ in murine molars." Developement. 122. 1243-1251 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Hiranuma: "Changes in the irradiation sensitivity of cultured growth palte chondrocytes during ctyodifferentiation." Oral Radiology. in press (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Enomoto-Iwamoto: "Involvement of α5β1 integrin in matrix interactions and proliferation of chondrocytes." J.Bone Miner.Res.in press (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Nakamura: "Sonic hedgefog synergistically promotes osteoblast differentiation with BMP-2." Exp.Cell Res.in press (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.J.Tabata: "Expression of cytokeratin 14 in ameloblast-lineage cells of the developing tooth of rat both in vivo and in vitro." Archs.oral Biol.(1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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