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Elucidation of DNA methylation status of bovine spermatozoa related to embryo development after in vitro fertilization and conception after artificial insemination

Research Project

Project/Area Number 16K07999
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Animal production science
Research InstitutionNational Agriculture and Food Research Organization

Principal Investigator

TAKEDA Kumiko  国立研究開発法人農業・食品産業技術総合研究機構, 畜産研究部門家畜育種繁殖研究領域, 上級研究員 (60414695)

Co-Investigator(Kenkyū-buntansha) 小林 栄治  国立研究開発法人農業・食品産業技術総合研究機構, 畜産研究部門, ユニット長 (00186727)
Research Collaborator WATANABE shinya  
HOSHINO yoichiro  
NISHINO kagetomo  
IMAI akira  
ADACHI hiromichi  
IWAO ken  
KANEDA masahiro  
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywordsウシ / 精子 / DNAメチル化 / 受胎性 / 体外受精 / 月齢変化 / メチル化 / マイクロアレイ / 畜産学 / 繁殖
Outline of Final Research Achievements

To evaluate the novel methods for the prediction of bull fertility based on epigenome information, we analyzed the genome-wide DNA methylation profiles of bull spermatozoa using a human DNA methylation microarray. We detected differentially methylated regions (DMRs) concerning embryonic developmental ability after in vitro fertilization (IVF%), conception rate after artificial insemination (AI%), and age-related changes. Using combined bisulfite restriction analysis (COBRA), nine DMRs were confirmed to exhibit significant differences in their age-dependent methylation levels. One DMR was detected to correlate with IVF%, and another one DMR was detected to correlate with AI%. Further studies are required to use these DMRs as epigenetic biomarkers for predicting semen or bull fertility.

Academic Significance and Societal Importance of the Research Achievements

ウシの精液性状検査による評価結果が良好でも人工授精後の受胎性の低い精液が存在しており、新たな評価指標が求められる。本研究ではエピゲノム情報を取り入れたウシ精液の新たな評価法の開発のため、ヒト用メチル化解析マイクロアレイを利用した網羅的メチル化解析から受胎性の異なるグループ間のメチル化プロファイルの解明や簡易評価指標となるメチル化可変部位の検出を行った。また、加齢変化を示す複数のメチル化可変領域のメチル化率から月齢推定がある程度可能であることから、今後、受胎性に関連するメチル化可変領域を複数特定することで、凍結精液の受胎性の事前評価の指標となることが期待される。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (12 results)

All 2019 2018 2017 2016

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (10 results) (of which Int'l Joint Research: 3 results)

  • [Journal Article] Age-related changes in DNA methylation levels at CpG sites in bull spermatozoa and <i>in vitro</i> fertilization-derived blastocyst-stage embryos revealed by combined bisulfite restriction analysis2019

    • Author(s)
      ①Kumiko Takeda, Eiji Kobayashi, Kagetomo Nishino, Akira Imai, Hiromichi Adachi, Yoichiro Hoshino, Satoshi Akagi, Masahiro Kaneda, Shinya Watanabe
    • Journal Title

      Journal of Reproduction and Development

      Volume: 65 Issue: 4 Pages: 305-312

    • DOI

      10.1262/jrd.2018-146

    • NAID

      130007689001

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Differentially methylated CpG sites in bull spermatozoa revealed by human DNA methylation arrays and bisulfite analysis2017

    • Author(s)
      Kumiko Takeda, Eiji Kobayashi, Satoshi Akagi, kagetomo Nishino, Masahiro Kaneda, Shinya Watanabe
    • Journal Title

      Journal of Reproduction and Development

      Volume: 63 Issue: 3 Pages: 279-287

    • DOI

      10.1262/jrd.2016-160

    • NAID

      130007314612

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 体外受精胚発生率に関連しうる牛精子核DNAのメチル化可変部位の探索2019

    • Author(s)
      武田久美子,小林栄治,今井昭,佐藤伸哉,西野景知,安達広通,岩尾健,赤木悟史,金田正弘,渡邊伸也
    • Organizer
      日本畜産学会第125回大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 体外受精後の胚における牛精子核DNAメチル化可変部位のメチル化状態2018

    • Author(s)
      武田久美子, 小林栄治, 星野洋一郎, 安達広通, 今井昭, 西野景知, 岩尾健, 金田正弘, 渡邊伸也
    • Organizer
      INRA-NARO Joint Symposium: Cutting-edge Research on Animal Breeding and Reproduction
    • Related Report
      2018 Annual Research Report
  • [Presentation] 牛精子核DNAのメチル化可変部位と凍結精液受胎率との関連性の検討2018

    • Author(s)
      武田久美子、小林栄治、星野洋一郎、安達広通、今井昭、西野景知、岩尾健、金田正弘、渡邊伸也
    • Organizer
      日本畜産学会第124回大会
    • Related Report
      2017 Research-status Report
  • [Presentation] Age-dependent changes of DNA methylation levels in bull spermatozoa2018

    • Author(s)
      Kumiko Takeda
    • Organizer
      INRA-NARO Joint Symposium: Cutting-edge Research on Animal Breeding and Reproduction
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 牛精液凍結時の還元型Lグルタチオンの添加が体外受精胚発生率に与える効果2017

    • Author(s)
      武田久美子・小林栄治・赤木悟史・渡邊伸也
    • Organizer
      日本畜産学会122回大会
    • Place of Presentation
      神戸大学(兵庫県・神戸市)
    • Year and Date
      2017-03-27
    • Related Report
      2016 Research-status Report
  • [Presentation] ヒトメチル化チップを用いたウシCpGサイトの網羅的メチル化解析2017

    • Author(s)
      小林栄治・武田久美子
    • Organizer
      日本畜産学会122回大会
    • Place of Presentation
      神戸大学(兵庫県・神戸市)
    • Year and Date
      2017-03-27
    • Related Report
      2016 Research-status Report
  • [Presentation] Age-related changes in DNA methylation levels of CpG sites in bull spermatozoa and IVF embryos.2017

    • Author(s)
      Kumiko Takeda, Eiji Kobayashi, Kagetomo Nishino, Akira Imai, Hiromichi Adachi, Yoichiro Hoshino, Satoshi Akagi, Masahiro Kaneda, Shinya Watanabe
    • Organizer
      4th World Congress of Reproductive Biology
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 人工授精後の受胎率に関連する牛精子核DNAのメチル化可変部位の検出2017

    • Author(s)
      武田久美子、小林栄治、星野洋一郎、安達広通、今井昭、西野景知、岩尾健、金田正弘、渡邊伸也
    • Organizer
      日本受精卵移植関連合同研究会
    • Related Report
      2017 Research-status Report
  • [Presentation] 繁殖供用開始期の雄牛精子核DNAの特定メチル化可変部位におけるメチル化状態の差異2016

    • Author(s)
      武田久美子・小林栄治・西野景知・今井昭・金田正弘・渡邊伸也
    • Organizer
      日本繁殖生物学会109回大会
    • Place of Presentation
      麻布大学(神奈川県・相模原市)
    • Year and Date
      2016-09-12
    • Related Report
      2016 Research-status Report
  • [Presentation] Differentially methylated CpG sites in bull sperm DNA revealed by human DNA methylation arrays2016

    • Author(s)
      Kumiko Takeda, Eiji Kobayashi, Satoshi Akagi, Masahiro Kaneda, Shinya Watanabe
    • Organizer
      第17回アジア・大洋州畜産国際会議(AAAP2016)
    • Place of Presentation
      福岡産業大学(福岡県・福岡市)
    • Year and Date
      2016-08-22
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research

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Published: 2016-04-21   Modified: 2020-03-30  

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