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Visualization and quantification of gene expression involved in pain transmission and regulation with molecular probes

Research Project

Project/Area Number 07557010
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field General pharmacology
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

SATOH Masamichi  Kyoto University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (80025709)

Co-Investigator(Kenkyū-buntansha) MINAMI Masabumi  Kyoto University, Faculty of Pharmaceutical Sciences, Instructor, 薬学部, 助手 (20243040)
KANEKO Shuji  Kyoto University, Faculty of Pharmaceutical Sciences, Associate professor, 薬学部, 助教授 (60177516)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥13,600,000 (Direct Cost: ¥13,600,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥12,300,000 (Direct Cost: ¥12,300,000)
Keywordsin situ hybridization / mRNA / double staining / opioid receptor / pain / digoxigenin / image analysis
Research Abstract

For the purpose to increase the sensitivity of an in situ hybridization technique with nonradioactive probes, we examined the several methods for nonradioactive labeling of RNA probes. Simiar sensitivities were obtained between digoxigenin- and fluorescein-labeled antisense RNA probes to detect the target mRNA,but the background signals were lower in the results with digoxigenin-labeled probe than with fluorescein-labeled one. For an analysis of the results of double in situ hybridization histochemistry, we developed an apparatus to simultaneously obtain both bright- and dark-field images, which were transferred to a personal computer and analyzed using a software for image analysis, NIH image. Coexistence of the mu-opioid receptor mRNA with the mRNA for preprotachykinin A (PPTA), a precursor protein of substance P,in the rat dorsal root ganglia was examined by this system, and we could quantitatively analyze the expression of mu-opioid receptor mRNA,which was detected by a ^<35>S-labeled probe, specifically in the cells expressing PPTA mRNA,which was visualized by a digoxigenin-labeled probe. Furthermore, we developed a method to analyze the of film-autoradiogram, and showed that expressions of mu-and kappa-, but not delta-, opioid receptor mRNAs were enhanced in the superfacial layrs of spinal dorsal horn of the arthritic rats.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Minami, M.: "Double in situ hybridization study on co-existence of μ-,δ-and κ-opioid receptor mRNAs with preprotachykinin A mRNA in the rat dorsal root ganglia." Mol. Brain Res.30. 203-210 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Maekawa, K.: "Expression of μ-and κ-,but not δ-,opioid receptor mRNAs are enhanced in the spinal dorsal horn of the arthritic rats." Pain. 64. 365-371 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yabuuchi, K.: "Induction of interleukin-1β mRNA in the hypothalamus following subcutaneous injections of fromalin into the rat hind paws." Neurosci. Lett.207. 109-112 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Minami, M.: "Double in situ hybridization study on coexistence of mu-, delta- and kappa-opioid receptor mRNAs with preprotachykinin A mRNA in the rat dorsal root ganglia." Mol.Brain.Res.30. 203-210 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Maekawa, K.: "Expression of mu- and kappa-, but not delta-, opioid receptor mRNAs are enhanced in the spinal dorsal horn of the arthritic rats." Pain.64. 365-371 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yabuuchi, K.: "Induction of interleukin-1beta mRNA in the hypothalamus following subcutaneous injections of formalin into the rat hind paws." Neurosci.Lett.207. 109-112 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Minami,M.: "Double in situ hybridization study on co-exsitence of μ-,δ-and κ-opioid receptor mRNAs with preprotachykinin A mRNA in the rat dorsal rootx gxanglia." Mol.Brain Res.30. 203-210 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Maekawa,K.: "Expression of μ-,and κ-,but not δ-,opioid receptor mRNAs are enhanced in the spinal dorsal horn of the arthritic rats." Pain. 64. 365-371 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yabuuchi,K.: "Induction of interleukin-1β mRNA in the hypothalmus following subcuxtaneous inuectxions of formalin into the rat hind paws." Neuroxsci.Lett.207. 109-112 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Minami, M.: "Double in situ hybridization study on co-existence of μ-, δ- and κ-opioid receptor mRNAs with preprotachykinin A mRNA in the rat dorsal root ganglia." Mol. Brain Res.30. 203-210 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Maekawa, K.: "Expression of μ- and κ-, but not δ-, opioid receptor mRNA s are enhanced in the spinal dorsal horn of the arthritic rats." Pain. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Yabuuchi, K.: "Induction of interleukin-1β mRNA in the hypothalamus following subcutaneous injections of formalin into the rat hind paws." Neurosci. Lett.(in press).

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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