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Is it possible to separate male and female chickens? -Through the analysis of the avian sex determination mechanism-

Research Project

Project/Area Number 19H03107
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 42010:Animal production science-related
Research InstitutionHiroshima University

Principal Investigator

Horiuchi Hiroyuki  広島大学, 統合生命科学研究科(生), 教授 (80243608)

Project Period (FY) 2019-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥16,900,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥3,900,000)
Fiscal Year 2022: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2021: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2020: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2019: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Keywordsニワトリ / 性決定 / DMRT1 / HEMGN / 始原生殖細胞 / PGC / 精子分化 / 雄性決定 / Dmrt1 / ゲノム編集
Outline of Research at the Start

本研究課題では,鳥類であるニワトリの性を決定するマスター遺伝子ではないかと考えられているDmrt1遺伝子に着目し,本遺伝子をゲノム編集技術によりノックアウトする。本実験により,Dmrt1遺伝子ノックアウトのヘテロ接合体(Dmrt1+/-)やホモ接合体(Dmrt1-/-)を作出し,その生殖巣の特徴や周辺遺伝子の発現に与える影響を解析することで,ニワトリにおける性決定のメカニズムを明らかにする。得られた知見をもとに,産業面で極めて重要なニワトリにおいて,雌雄の産み分けが可能かどうかを検討し,応用展開をはかる。

Outline of Final Research Achievements

In this research, I focused on the DMRT1 and HEMGN genes, which are thought to be the master genes that determine the sex of the chicken. At 7.5 days of incubation, neither normal PGCs nor DMRT1-knockout PGCs showed any change in distribution and number. In 14-day-incubated embryos, the proportion of Nanog-expressing PGCs was high in DMRT1-knockout PGCs. Conversely, spermatogonial stem cell markers were low in DMRT1-knockout PGCs. Knockout analysis of HEMGN showed abnormalities in the hematopoietic system in females, and yellow discoloration of the liver, atrophy of the fallopian tubes, and underdevelopment of follicles were observed. In HEMGN heterozygous knockout males, gene expression of HEMGN and DMRT1 was halved at 8.5 days of incubation. SOX9, which is expressed in male gonads following HEMGN and DMRT1, decreased slightly, and AMH, which inhibits Mullerian duct differentiation, did not change. These results indicate direct and indirect interactions between DMRT1 and HEMGN.

Academic Significance and Societal Importance of the Research Achievements

本研究課題は,現在,世界中で問題となっている養鶏における動物福祉に資するものである。養鶏の現場では,採卵が目的の場合はオスの雛を,食肉が目的の場合はメスの雛の一部が淘汰されている。EU各国では既に,雛の殺処分を禁止する動きが強まっており,早急な対策が求められている。本研究では,将来,ニワトリでの雌雄の産み分け技術に利用可能な知見を得るための分子生物学的な解析を行い,特定の遺伝子がニワトリの精子形成に重要であり,またその相互作用を明らかにした。これらの知見の集積により,近い将来,ニワトリで雌雄の産み分けが可能になるかもしれない。

Report

(5 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • Research Products

    (40 results)

All 2023 2022 2021 2020 2019 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 4 results) Presentation (24 results) (of which Int'l Joint Research: 2 results,  Invited: 4 results) Book (5 results) Remarks (4 results)

  • [Journal Article] Fate Decisions of Chicken Primordial Germ Cells (PGCs): Development, Integrity, Sex Determination, and Self-Renewal Mechanisms2023

    • Author(s)
      Kennosuke Ichikawa, Hiroyuki Horiuchi
    • Journal Title

      Genes

      Volume: 14 Issue: 3 Pages: 612-612

    • DOI

      10.3390/genes14030612

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genome editing in chickens2022

    • Author(s)
      Kennosuke Ichikawa, Mei Matsuzaki, Ryo Ezaki, Hiroyuki Horiuchi
    • Journal Title

      Gene and Genome Editing

      Volume: 3-4 Pages: 1-6

    • DOI

      10.1016/j.ggedit.2022.100015

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Prediction of sex-determination mechanisms in avian primordial germ cells using RNA-seq analysis2022

    • Author(s)
      Kennosuke Ichikawa, Yoshiaki Nakamura, Hidemasa Bono, Ryo Ezaki, Mei Matsuzaki & Hiroyuki Horiuchi
    • Journal Title

      Scientific Reports

      Volume: 12 Issue: 1 Pages: 13528-13528

    • DOI

      10.1038/s41598-022-17726-7

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Targeted Knock-in of a Fluorescent Protein Gene into the Chicken Vasa Homolog Locus of Chicken Primordial Germ Cells using CRIS-PITCh Method2022

    • Author(s)
      Ryo Ezaki, Kennosuke Ichikawa, Mei Matsuzaki, Hiroyuki Horiuchi
    • Journal Title

      The Journal of Poultry Science

      Volume: 59 Issue: 2 Pages: 182-190

    • DOI

      10.2141/jpsa.0210067

    • NAID

      130008093959

    • ISSN
      1346-7395, 1349-0486
    • Related Report
      2022 Annual Research Report 2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Knock-in of the duck retinoic acid-inducible gene I (RIG-I) into the Mx gene in DF-1 cells enables both stable and immune response-dependent RIG-I expression2021

    • Author(s)
      Kennosuke Ichikawa, Yuzuha Motoe, Ryo Ezaki, Mei Matsuzaki, Hiroyuki Horiuchi
    • Journal Title

      Biochemistry and Biophysics Reports

      Volume: 27 Pages: 101084-101084

    • DOI

      10.1016/j.bbrep.2021.101084

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] An improved protocol for stable and efficient culturing of chicken primordial germ cells using small-molecule inhibitors2020

    • Author(s)
      Ezaki Ryo、Hirose Fumiya、Furusawa Shuichi、Horiuchi Hiroyuki
    • Journal Title

      Cytotechnology

      Volume: 72 Issue: 3 Pages: 397-405

    • DOI

      10.1007/s10616-020-00385-9

    • NAID

      120007097011

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Comparison of sex determination mechanism of germ cells between birds and fish: Cloning and expression analyses of chicken forkhead box L3-like gene.2019

    • Author(s)
      Kennosuke Ichikawa, Ryo Ezaki, Shuichi Furusawa, Hiroyuki Horiuchi
    • Journal Title

      Developmental Dynamics

      Volume: 248 Issue: 9 Pages: 826-836

    • DOI

      10.1002/dvdy.67

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] ニワトリマクロファージ様細胞HD11において強力な抗炎症応答を示す CpGオリゴDNA の選抜2022

    • Author(s)
      市川健之助、松崎芽衣、江崎僚, 堀内浩幸、山本祥也
    • Organizer
      第46回鳥類内分泌研究会
    • Related Report
      2022 Annual Research Report
  • [Presentation] ニワトリ生殖細胞の分化過程に関する研究2022

    • Author(s)
      山脇まゆ子、江崎僚、松崎芽衣、堀内浩幸
    • Organizer
      第46回鳥類内分泌研究会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 細胞周期阻害剤を用いたニワトリ細胞への遺伝子導入に関する研究2022

    • Author(s)
      越智勇太、渡邊天海、池谷亮、江崎僚、松崎芽衣、堀内浩幸
    • Organizer
      第46回鳥類内分泌研究会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 分化したニワトリ始原生殖細胞の性状解析 -鳥類始原生殖細胞の新規培養系の確立に向けて-2022

    • Author(s)
      市川 健之助, 元榮 柚花, 江崎 僚, 松崎 芽衣, 堀内 浩幸
    • Organizer
      第45回日本分子生物学会年会
    • Related Report
      2022 Annual Research Report
  • [Presentation] ニワトリエピブラスト培養細胞は多能性幹細胞になりえるのか2022

    • Author(s)
      梶原 亮太, 市川 健之助, 江崎 僚, 松崎 芽衣, 堀内 浩幸
    • Organizer
      第45回日本分子生物学会年会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Construction of an Evaluation System for Chicken Egg Bioreactor Using Cultured Chicken Cells2022

    • Author(s)
      Ryota Kajihara, Kenosuke Ishikawa, Ryo Ezaki, Mei Matsuzaki, Hiroyuki Horiuchi
    • Organizer
      International Seminar 2022:Front-line of genome editing technology
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Genetic Engineering in Chicken Primordial Germ Cells2022

    • Author(s)
      Tenkai Watanabe, Kenosuke Ishikawa, Ryo Ezaki, Mei Matsuzaki, Hiroyuki Horiuchi
    • Organizer
      International Seminar 2022:Front-line of genome editing technology
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ゲノム編集とデータサイエンスを活用した新たな家禽の利用方法と未来2022

    • Author(s)
      堀内 浩幸
    • Organizer
      第130回有機デバイス研究会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Knock-in strategy by genome editing in chicken cells2022

    • Author(s)
      Tenkai Watanabe, Yuta Ochi, Ryo Ezaki, Mei Matsuzaki, Hiroyuki Horiuchi
    • Organizer
      日本ゲノム編集学会第7回大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Construction of an evaluation system for chicken egg bioreactor using cultured chicken cells2022

    • Author(s)
      Ryota Kajihara, Ryo Ezaki, Mei Matsuzaki, Hiroyuki Horiuchi
    • Organizer
      日本ゲノム編集学会第7回大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] ゲノム編集技術が食料生産を変える!2022

    • Author(s)
      堀内 浩幸
    • Organizer
      広島バイオテクノロジー推進協議会第4回Web講演会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] ニワトリ始原生殖細胞の体細胞分化に関する研究2021

    • Author(s)
      市川健之助・元榮柚花・江崎僚・松崎芽衣・堀内浩幸
    • Organizer
      日本家禽学会2022年度春季大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ニワトリ培養細胞のheparinによる遺伝子導入効率の低下現象に関する研究2021

    • Author(s)
      越智勇太, 渡邊天海, 市川健之助, 江崎僚, 松崎芽衣, 堀内浩幸
    • Organizer
      第45回鳥類内分泌研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ニワトリPGCsにおける遺伝子工学的な特徴2021

    • Author(s)
      渡邊天海,越智勇太,市川健之助,江崎僚,松崎芽衣,堀内浩幸
    • Organizer
      第45回鳥類内分泌研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 鳥類アデノ随伴ウイルスベクターのウズラ胚に対する感染能評価2021

    • Author(s)
      山西菜々子,江崎僚,笹浪知宏,松崎芽衣,堀内浩幸
    • Organizer
      第45回鳥類内分泌研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ニワトリ生殖細胞の分化過程に関する研究2021

    • Author(s)
      山脇まゆ子,江崎 僚, 松崎芽衣, 堀内浩幸
    • Organizer
      第44回 日本分子生物学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ゲノム編集の今とこれから2021

    • Author(s)
      堀内浩幸
    • Organizer
      NACS中国支部研修講座
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] ニワトリ生殖細胞の分化過程に関する研究2021

    • Author(s)
      山脇まゆ子,江崎 僚, 松崎芽衣, 堀内浩幸
    • Organizer
      日本ゲノム編集学会第6回大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ゲノム編集を用いた家禽の品種改良技術の確立2021

    • Author(s)
      堀内浩幸
    • Organizer
      COI-NEXTxOPERAx卓越大学院 オンライン合同シンポジウム「TRINITY」
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 遺伝子発現プロファイルに基づく, ニワトリ始原生殖細胞の性分化機構の予測2020

    • Author(s)
      市川健之助、中村隼明、江崎僚、松崎芽衣、堀内浩幸
    • Organizer
      第44回鳥類内分泌研究会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 鳥類性決定関連遺伝子の機能解析2020

    • Author(s)
      桐山海里、山田智香子、江崎僚、松崎芽衣、堀内浩幸
    • Organizer
      第44回鳥類内分泌研究会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ゲノム編集技術を用いたニワトリ性決定機構の解明2020

    • Author(s)
      山脇まゆ子、江崎僚、松崎芽衣、堀内浩幸
    • Organizer
      第44回鳥類内分泌研究会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Analyses of germ-cell fate determination in chickens2019

    • Author(s)
      Hiroaki Matsumoto, Ryo Ezaki, Ryota Kajihara, Mei Matsuzaki, Shuichi Furusawa, Hiroyuki Horiuchi
    • Organizer
      4th Annual Meeting of the Japanese Society for Genome Editing
    • Related Report
      2019 Annual Research Report
  • [Presentation] Elucidation of mechanism of chicken sex determination using genome editing2019

    • Author(s)
      Yuki Okaza, Mayuko Ymawaki, Ryo Ezaki, Mai Matsuzaki, Shuichi Furusawa, Hiroyuki Horiuchi
    • Organizer
      4th Annual Meeting of the Japanese Society for Genome Editing
    • Related Report
      2019 Annual Research Report
  • [Book] ゲノム編集技術~実験上のポイント/産業利用に向けた研究開発動向と安全性周知2023

    • Author(s)
      堀内浩幸
    • Total Pages
      7
    • Publisher
      株式会社 情報機構
    • ISBN
      9784865022421
    • Related Report
      2022 Annual Research Report
  • [Book] つくると食べるをつなぐサイエンス2022

    • Author(s)
      堀内浩幸,松崎芽衣
    • Total Pages
      8
    • Publisher
      株式会社 恒星社厚生閣
    • ISBN
      9784769916857
    • Related Report
      2022 Annual Research Report
  • [Book] SDGsに向けた生物生産学入門2021

    • Author(s)
      1章14:ゲノム編集とは何か(堀内浩幸)
    • Total Pages
      244
    • Publisher
      共立出版
    • ISBN
      9784320058316
    • Related Report
      2021 Annual Research Report
  • [Book] 最新のゲノム編集技術と用途展開2021

    • Author(s)
      堀内浩幸,松崎芽衣,江崎僚(監修 山本卓)
    • Total Pages
      240
    • Publisher
      株式会社シーエムシー出版
    • ISBN
      9784781315867
    • Related Report
      2020 Annual Research Report
  • [Book] ゲノム編集実験スタンダード2019

    • Author(s)
      江崎僚, 松崎芽衣, 堀内浩幸
    • Total Pages
      386
    • Publisher
      羊土社
    • ISBN
      9784758122443
    • Related Report
      2019 Annual Research Report
  • [Remarks] 免疫生物学研究室

    • URL

      https://immunobi.hiroshima-u.ac.jp/

    • Related Report
      2022 Annual Research Report
  • [Remarks] ニワトリの始原生殖細胞における性決定機構の予測に成功 ―鳥類特有の性決定機構の解明に向けて―

    • URL

      https://www.hiroshima-u.ac.jp/news/72553

    • Related Report
      2022 Annual Research Report
  • [Remarks] 広島大学・大学院統合生命科学研究科・免疫生物学研究室

    • URL

      http://immunobi.hiroshima-u.ac.jp/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 教授に聞く

    • URL

      https://gsbstop.hiroshima-u.ac.jp/interview/ja/horiuchi.html

    • Related Report
      2020 Annual Research Report

URL: 

Published: 2019-04-18   Modified: 2024-01-30  

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