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Transcription factors involved in the gene transcription of calcium-regulating hormone

Research Project

Project/Area Number 18570058
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphology/Structure
Research InstitutionShizuoka University

Principal Investigator

SUZUKI Masakazu  Shizuoka University, Faculty of Science, Associate Professor (60280913)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥4,110,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsRainbow trout / Calcitonin / Ultimobranchial gland / Thyroidal C cells / Transcription factors
Research Abstract

Calcitonin (CT) inhibits bone resorption in mammals, and is used as a therapeutic medicine for the treatment of bone disorders. In mammals, this hormone is secreted by the C-cells scattered in the thyroid, but as for the other vertebrates, CT-producing cells form the ultimobranchial gland. However, little is known about the molecular mechanisms intensifying CT gene expression in these cells.
In the first experiment, I have characterized mRNAs expressed in the ultimobranchial gland of rainbow trout (Oncorhynchus mykiss) by suppression-subtractive hybridization. Sequence analysis of 110 cDNA fragments indicated the existence of three types of keratins, Is (88), He (E3), and IIs (S2), and ictacalcin, a calcium-binding protein reported to be expressed in the epidermis in mammals, suggesting that the CT cell has characteristics of the epithelial cell. Cytoskeletal protein 4.1G was also identified. Furthermore, I found a secretory glycoprotein, clusterin, which is implicated in various physio … More logical processes in other animals. RT-PCR analysis showed that clusterin gene is expressed in all the tissues examined in rainbow trout, with the highest level in the ultimobranchial gland. Immunofluorescene staining with an anti-clusterin antibody revealed positive labels over the ultimobranchial gland. Intense immunoreativity was observed along the apical membrane of CT cells facing the follicular lumen, suggesting that clusterin accumulates in the follicles of ultimobranchial gland. Positive labels were also detected along the apical membrane of the follicular epithelial cells of the thyroid, at goblet cells in the intestine, and at mucous cells in the gills.
As for transcription factors expressed in the ultimobranchial gland, Nkx2.1d, Paxl, and FoxF1 had been identified by cDNA cloning. It was recently reported that FoxA2 (HNF-3beta) is expressed in mammalian CT-producing cell lines, CA77, and increases transcriptional activity of rat CT/CT gene-related peptide (CGRP) gene. However, that effect of FoxA2 does not seem to be strong enough. In the second experiment, therefore, I tried cloning the cDNAs for winged helix/forkhead transcription factor Foxa (HNF-3) or similar factors from the ultimobranchial gland of rainbow trout. The cDNA encoding FoxA2 was isolated, and RT-PCR analysis showed the expression of FoxA2 mRNA in the UB, liver, and intestine. FoxA2 is considered to exist in the parenchymal cells of UB.To investigate whether FoxA2 activate stranscription from the CT promoter, I further cloned a DNA fragment encompassing the 2 kbp upstream region and 5'-noncoding region of the exon I of salmon-type CT-Ib gene from the rainbow trout, using PCR. In addition, I cloned a 3.8 kbp DNA fragment ranging from the 3'-noncoding region of exon IV to downstream region of CT/CGRP gene from medaka, using PCR. I inserted these DNA fragments into a luciferase reporter vector, and performed luciferase assay. However, transcriptional activity was not increased by co-transfection of FoxA1 and/or FoxA2 with the luciferase vector. Less

Report

(2 results)
  • 2007 Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (16 results)

All 2007 2006

All Journal Article (8 results) (of which Peer Reviewed: 1 results) Presentation (8 results)

  • [Journal Article] Functional analysis of Nkx2.1 and Pax9 for calcitoningene transcription.2007

    • Author(s)
      M. Suzuki, N. Katagiri, M. Ueda, & S. Tanaka
    • Journal Title

      Gen. Comp. Endocrinol. 152

      Pages: 259-266

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Functional analysis of Nkx2.1 and Pax9 for calcitonin gene transcription2007

    • Author(s)
      M. Suzuki, N. Katagiri, M. Ueda, S. Tanaka
    • Journal Title

      Gen. Comp. Endocrinol 152

      Pages: 259-266

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Functional analysis of Nkx2.1 and Pax9 for calcitonin gene transcription.2007

    • Author(s)
      Suzuki M, Katagiri N, Ueda M, Tanaka S
    • Journal Title

      Gen Comp Endocrinol (In press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] 甲状腺の分化に関連する遺伝子Nkx2.1の分子進化2006

    • Author(s)
      鈴木雅一、田中滋康
    • Journal Title

      生体の科学 57

      Pages: 508-509

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Suppression subtractive hybridization法と内分泌学研究への応用2006

    • Author(s)
      鈴木雅一、田中滋康
    • Journal Title

      日本比較内分泌学会ニュース 123

      Pages: 27-31

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Molecular evolution of Nkx2.1 involved in thyroid formation2006

    • Author(s)
      M. Suzuki, S. Tanaka
    • Journal Title

      Seitai No Kagaku 57

      Pages: 508-509

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Suppression subtractive hybridization and its application to endocrine research2006

    • Author(s)
      M. Suzuki, S. Toki
    • Journal Title

      Comp. Endocrinol 123

      Pages: 27-31

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] 甲状腺の分化に関連する遺伝子Nkx2.1の分子進化2006

    • Author(s)
      鈴木雅一, 田中滋康
    • Journal Title

      生体の科学 生物進化の分子マップ 57

      Pages: 508-509

    • Related Report
      2006 Annual Research Report
  • [Presentation] ニジマスにおけるクラスタリンの分布に関する分子組織学的解析2007

    • Author(s)
      福井亮司、中倉敬、田中滋康、鈴木雅一
    • Organizer
      第32回日本比較内分泌学会大会
    • Place of Presentation
      日光
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Molecular histological analysis of the distribution of clasterin in the rainbow trout2007

    • Author(s)
      Fukui, R., Nakakura, T., Tanaka, S., Suzuki, M
    • Organizer
      31st Conference of Japan Comparative Endocrinology
    • Place of Presentation
      Nikko
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Transcription factors expressed in calcitonin-producing cells and their functions2006

    • Author(s)
      Suzuki, M., Katagiri, N., Toki, S., Ueda, M., Tanaka, S
    • Organizer
      23rd conference of European Comparative Endocrinology
    • Place of Presentation
      Manchester. UK
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ニジマス鯉後腺で発現しているmRNAの解析2006

    • Author(s)
      土岐晋吾、上田誠、田中滋康、鈴木雅一
    • Organizer
      第77回日本動物学会大会
    • Place of Presentation
      松江
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ウシガエル副甲状腺で発現しているmRNAの解析2006

    • Author(s)
      上田 誠, 田中滋康, 鈴木雅一
    • Organizer
      第31回日本比較内分泌学会大会
    • Place of Presentation
      札幌
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Transcription factors expressed in calcitonin-producing cells and their functions2006

    • Author(s)
      Suzuki, M., Katagiri, N., Toki, S., Ueda, M., Tanaka, S
    • Organizer
      23rd Conference of European Comparative Endocrinology
    • Place of Presentation
      Manchester, UK
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Analysis of mRNA expressed in the ultimobranchial gland2006

    • Author(s)
      Told, S., Ueda, M., Tanaka, S., Suzuki, M
    • Organizer
      77rd Conference of Japan Zoological Society
    • Place of Presentation
      Matsue
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Analysis of mRNA expressed in the parathyroid gland2006

    • Author(s)
      Ueda, M., Tanaka, S., Suzuki, M
    • Organizer
      31st Conference of Japan Comparative Endocrinology
    • Place of Presentation
      Sapporo
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2006-04-01   Modified: 2016-04-21  

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