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1996 Fiscal Year Final Research Report Summary

Development of a new method for detecting gene expression in a single cell and its medical application.

Research Project

Project/Area Number 07557015
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field Pathological medical chemistry
Research InstitutionTohoku University

Principal Investigator

SHIBAHARA Shigeki  Tohoku University School of Medicine, Professor, 医学部, 教授 (70206142)

Co-Investigator(Kenkyū-buntansha) YASUMOTO Kenichi  Tohoku University School of Medicime, Assistant Professor, 医学部, 助手 (90241629)
TAKAHASHI Kazuhiro  Tohoku University School of Medicine, Lecturer, 医学部, 講師 (80241628)
SHIBATA Koushi  Pola R & D Laboratories, Dermal Research Department, Staff Investigator, 医薬品研究所, 副主任研究員
Project Period (FY) 1995 – 1996
Keywordstranscription factor / melanin biosynthesis / heme catabolism / tyrosinase / heme oxygenase / cell differentiation / MITF
Research Abstract

We have successfully developed a new luciferase assay technology that enables the expression of pigment cells, transfected with the fusion gene containing the firefly luciferase gene, to be captured on photographic images. The light discharged by the luciferase-mediated reaction can be quantified by chemical luminescence measurement. Using the photocounting camera and an efficient transfection method, we were able to detect the promoter activity of the tyrosinase gene in human melanocytes. Furthermore, we have obtained the following results as summarized below. 1. The three cis-regulatory elements of the human tyrosinase gene have been identified, and each of these elements contains one copy of a CATGTG motif. We showed that each CATGTG motif is required for pigment cell-specific transcription. 2. In the course of studying the regulation of the heme oxygenase-1 gene expression during macrophage differentiation, we showed the presence of a putative silencer sequence, repressing a function of the heat shock element, which is located in the 5^1-flanking region of the human heme oxygenase-1 gene. 3. Three types of nitric oxide (NO) donors increased the expression levels of heme oxygenase-1 mRNA,suggesting a possible coordinated regulation of heme oxygenase-1 and NO synthase mRNAs. 4. Both heme oxygenase-1 and inducible NO synthase mRNAs were overexpressed in human primary brain tumors compared to the normal brain tissues. 5. The choroid plexus probuces cerebrospinal fluid which plays an important role not only in the mechanical support of the brain but also in the chemical homeostasis of the brain. We found that adrenomedullin, a potent vasodilator peptide, is produced and secreted by cultured choroid plexus carcinoma cells, a rare neoplasm derived from the epithelium of the choroid plexus.

  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Yasumoto K.et.al.: "Transcriptional Activation of the Melanocyte-Specific Genes by the Human Homolog of the Mouse Microphthalmia Protein." J.Biochem.(Tokyo). 118. 874-881 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takimoto H.et.al.: "MAT-1,a Monoclonal Antibody that Specifically Recognizes Human Tyrosinase." J.Invest.Dermatol.105. 764-768 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Muraosa Y.et.al.: "cDNA Cloning of a Novel Protein with the Two Zinc-Finger Domains that may Function as a Transcription Factor for the Human Heme Oxygenase-1 Gene." Eur.J.Biochem.235. 471-479 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fuse N.et.al.: "Identification of a Melanocyte-Type Promoter of the Microphthalmia-Associated Transcription Factor Gene." Biochem.Biophys.Res.Commun.219. 702-707 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okinaga S.et.al.: "Regulation of Human Heme Oxygenase-1 Gene Expression under Thermal Stress." Blood. 87. 5074-5084 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takahashi K.et.al.: "Expression of Heme Oxygenase Isozyme mRNAs in the Human Brain and Induction of Heme Oxygenase-1 by Nitric Oxide Donors." J.Neurochem. 67. 482-489 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hara E.et.al.: "Expression of Heme Oxygenase and Inducible Nitric Oxide Synthase mRNA in Human Brain Tumors" Biochem.Biophys.Reo.Commun.224. 153-158 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki H.et al.: "Differential Tissue-Specific Expression of Neurofibromin Isoform mRNAs in Rat." J.Biochem.(Tokyo). 120. 1048-1054 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasumoto K.et al.: "Functional Analysis of Microphthalmia-Associated Transcription Factor in Pigment Cell-specific Transcription of the Human Tyrosinase Family Genes." J.Biol.Chem.272. 503-509 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takahashi K.et al.: "Production and Secretion of Adrenomedullin by Cultured Choroid Plexus Carcinoma Cells." J.Neurochem.68. 726-731 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasumoto K. & Shibahara S.: "Molecular Cloning of a cDNA Encoding a Human TFEC Isoform,a Newly Identified Transcriptional Regulator" Biochim.Biophys.Acta. (印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasumoto, K., Mahalingam, H., Suzuki, H., Yoshizawa, M., Yokoyama, K.and Shibahara, S.: "Transcriptional Activation of the Melanocyte-Specific Genes by the Human Homolog of the Mouse Microphthalmia Protein." J.Biochem. (Tokyo). 118. 874-881 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takimoto, H., Suzuki, S., Masui, S., Shibata, K., Tomita, Y., Shibahara, S.and Nakano, H: "MAT-1, a Monoclonal Antibody that Specifically Recognizes Human Tyrosinase." J.Invest. Dermatol. 105. 764-768 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Muraosa, Y., Takahashi, K., Yoshizawa, M.and Shibahara, S: "cDNA Cloning of a Novel Protein with the Two Zinc-Finger Domains that may Function as a Transcription Factor for the Human Heme Oxygenase-1 Gene." Eur. J.Biochem.235. 471-479 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fuse, N., Yasumoto, K., Suzuki, H., Takahashi, K.and Shibahara, S: "Identification of a Melanocyte-Type Promoter of the Microphthalmia-Associated Transcription Factor Gene." Biochem. Biophys. Res. Commum. 219. 702-707 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Okinaga, S., Takahashi, K., Takeda, K., Yoshizawa, M., Fujita, H., Sasaki, H.and Shibahara, S: "Regulation of Human Heme Oxygenase-1 Gene Expression under Thermal Stress." Blood. 87. 5074-5084 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, K., Hara, E., Suzuki, H., Sasano, H.and Shibahara, S: "Expression of Heme Oxygenase Isozyme mRNAs in the Human Brain and Induction of Heme Oxygenase-1 by Nitric Oxide Donors." J.Neurochem. 67. 482-489 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hara, E., Takahashi, K., Tominaga, T., Kumabe, T., Kayama, T., Suzuki, H., Fujita, H., Yoshimoto, T., Shirato, K.and Shibahara, S: "Expression of Heme Oxygenase and Inducible Nitric Oxide Synthase mRNA in Human Brain Tumors." Biochem. Biophys. Res. Commun. 224. 153-158 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki, H., Takahashi, K., Yasumoto, K., Fuse, N.and Shibahara, S: "Differential Tissue-Specific Expression of Neurofibromin Isoform mRNAs in Rat." J.Biochem. (Tokyo). 120. 1048-1054 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasumoto, K., Yokoyama, K., Takahashi, K., Tomita, Y.and Shibahara, S: "Functional Analysis of Microphthalmia-Associated Transcription Factor in Pigment Cell-specific Transcription of the Human Tyrosinase Family Genes." J.Biol. Chem.272. 503-509 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, K., Satoh, F., Hara, E., Murakami, O., Kumabe, T., Tominaga, T., Kayama, T., Yoshimoto, T.and Shibahara, S: "Production and Secretion of Adrenomedullin by Cultured Choroid Plexus Carcinoma Cells." J.Neurochem. 68. 726-731 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasumoto, K.and Shibahara, S: "Molecular Cloning of a cDNA Encoding a Human TFEC Isoform, a Newly Identified Transcriptional Regulator." Biochim. Biophys. Acta. (in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-09  

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