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2006 Fiscal Year Final Research Report Summary

Experimental embryological study for induction of germ line cell through germ-line chimera in teleost fish.

Research Project

Project/Area Number 16380124
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General fisheries
Research InstitutionHokkaido University

Principal Investigator

YAMAHA Etsuro  Hokkaido Univ., Field Science Center for Northern Biosphere, Professor, 北方生物圏フィールド科学センター, 教授 (60191376)

Co-Investigator(Kenkyū-buntansha) ADACHI Shinji  Hokkaido Univ., Faculty of Fisheries Science, Professor, 大学院水産科学研究院, 教授 (40231930)
YOSHIZAKI Goro  Tokyo Univ.of Marine Science and Technology, Associate Professor, 海洋科学部, 助教授 (70281003)
HIRAI Toshiro  Teikyo Univ.of Sci. and Tech., Div.of Advance Science and Technology., Lecturer, 理工学部, 講師 (30238331)
Project Period (FY) 2004 – 2006
Keywordsgerm line chimera / primordial germ cell / PGC / xenogeneic chimera / germ plasm / nos 1 / vas
Research Abstract

This project was planned for development of foundational technology of surrogate production in teleost fish. The surrogate production in fish is a technique to get the gametes of some genotype through the gonad of other genotype. We got the following results in the past three years.
1)When artificial mRNA, combined with green fluorescence protein (GFP) and zebrafish nos1 3'UTR, was injected into fertilized eggs in herring, goldfish, loach, pearl danio, ice goby, ukigori goby, medaka, verfin flounder and eel, primordial germ cells (PGCs) were visualized during embryonic development.
2)Primordial germ cells (PGCs) were visualized during embryonic development in herring, goldfish, loach, pearl danio, ice goby, ukigori goby, medaka, verfin flounder and eel by injecting artificial chimeric mRNA combined with green fluorescence protein (GFP) and zebrafish nos1 3'UTR into fertilized eggs.
3)A method for single PGC transplantation was developed and named SPT method. In this method, visualized PGC … More s with GFP were isolated from early somitogenesis stage and transplanted into the blastulae in which differentiation of endogenous PGC was suppressed by injecting morpholino oligo (MO) nucleotide designed to maternally-supplied dead end (dnd) mRNA.
4)Fertilized eggs and viable offspring of pearl danio, inter-specifically different from zebrafish, were obtained from natural spawning of zebrafish parents developed from blastula to which PGC from pearl danio embryo were transplanted. Functional sperm of goldfish or loach belonging to a different genus or family from that of zebrafish, respectively, were also differentiated in the zebrafish host developed from blastulae to which PGC from each species were transplanted.
5)Androgenetic embryo between loach egg and goldfish sperm was lethal before epiboly. The blastomeres of this cybrid, however, were survived in the host embryos, when they were transplanted into the loach or goldfish blastulae. PGCs from this lethal cybrid were also detected in the host embryos of goldfish or loach.
6)Functional gametes were differentiated from PGCs isolated from cryopreserved genital anlage taken from hatched fry and transplanted peritoneal cavity of host fry. All rainbow trout offspring were also obtained from the triploid masu salmon of which PGC from rainbow trout were transplanted into peritoneal cavity of hatched fry.
7)In rainbow trout, functional gametes were differentiated from gonium, when they were isolated from developed gonads and transplanted into peritoneal cavity of hatched fry. Spermatogonia and oogonia were differentiated to egg and sperm, respectively, in the female and male host. Less

  • Research Products

    (38 results)

All 2007 2006 2005 2004

All Journal Article (38 results)

  • [Journal Article] A novel transforming growth factor-β superfamily member expressed in gonadal somatic cells enhances primordial germ cell and spermatogonial proliferation in rainbow trout (Oncorhynchus mykiss).2007

    • Author(s)
      Sawatari, E.他3名
    • Journal Title

      Dev. Biol. 301

      Pages: 266-275

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Generation of viable fish from cryopreserved primordial germ cells.2007

    • Author(s)
      Kobayashi, T.他3名
    • Journal Title

      Mol. Reprod. Dev. 74

      Pages: 207-213

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] 精巣の細胞から卵をつくる.魚類精原細胞の性的可塑性.2007

    • Author(s)
      吉崎悟朗
    • Journal Title

      BIO INDUSTRY 24

      Pages: 80-85

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] A novel transforming growth factor-β superfamily member expressed in gonadal somatic cells enhances primordial germ cell and spermatogonial proliferation in rainbow trout (Oncorhynchus mykiss).2007

    • Author(s)
      Sawatari, E., Shikina, S., Takeuchi, T., Yoshizaki, G.
    • Journal Title

      Dev.Biol. 301

      Pages: 266-275

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Generation of viable fish from cryopreserved primordial germ cells.2007

    • Author(s)
      Kobayashi, T., Takeuchi, Y., Takeuchi, T., Yoshizaki, G.
    • Journal Title

      Mol.Reprod.Dev. 74

      Pages: 207-213

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Temperature-dependent sex differentiation in goldfish : establishing the temperature-sensitive period and effect of constant and fluctuating water temperatures.2006

    • Author(s)
      Goto-Kazeto, R.
    • Journal Title

      Aquaculture 254

      Pages: 617-624

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Developmental stages and germ cell lineage of the loath (Misgurnus anguillicaudatus).2006

    • Author(s)
      Fujimoto, T.
    • Journal Title

      Zool. Sci. 23

      Pages: 977-989

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish.2006

    • Author(s)
      Okutsu, T.
    • Journal Title

      PNAS 103

      Pages: 2725-2729

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Manipulation of fish germ cells : visualization cryopreservation and transplantation.2006

    • Author(s)
      Okutsu, T.
    • Journal Title

      J. Reprod. Dev. 52

      Pages: 685-693

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Visualization of primordial germ cells in vivo using GFP-nosl 3'UTR mRNA2006

    • Author(s)
      Saito, T.
    • Journal Title

      Int. J. Dev. Biol. 50

      Pages: 691-700

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Drastic mortality in tetraploid induction results from the elevation of ploidy in masu salmon Oncorhynchus masou.2006

    • Author(s)
      Sakao, S.
    • Journal Title

      Aquaculture 252

      Pages: 147-160

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] 生殖細胞の異種間移植を利用した魚類養殖法.2006

    • Author(s)
      吉崎悟朗
    • Journal Title

      農林水産技術研究ジャーナル 29

      Pages: 57-60

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Green fluorescent protein labeling of primordial germ cells using a nontransgenic method and its application for germ cell transplantation in Salmonidae.2006

    • Author(s)
      Yoshizaki, G., Tago, Y., Takeuchi, Y., Sawatari, E., Kobayashi, T., Takeuchi, T.
    • Journal Title

      Biol.Reprod. 73

      Pages: 88-93

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Developmental stages and germ cell lineage of the loach (Misgurnus anguillicaudatus).2006

    • Author(s)
      Fujimoto, T., Kataoka, T., Sakao, S., Saito, T., Yamaha, E., Arai, K.
    • Journal Title

      Zool.Sci. 23

      Pages: 977-989

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Temperature-dependent sex differentiation in goldfish : establishing the temperature-sensitive period and effect of constant and fluctuating water temperatures.2006

    • Author(s)
      Goto-Kazeto, R., Abe, Y., Masai, M., Yamaha, E., Adachi, S., Yamauchi, K.
    • Journal Title

      Aquaculture 254

      Pages: 617-624

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish.2006

    • Author(s)
      Okutsu, T., Suzuki, K., Takeuchi, Y., Takeuchi, T., Yoshizaki, G.
    • Journal Title

      PNAS 103

      Pages: 2725-2729

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Manipulation of fish germ cells : visualization cryopreservation and transplantation.2006

    • Author(s)
      Okutsu, T., Yano, A., Nagasawa, K., Shikina, S., Kobayashi, T., Takeuchi, Y., Yoshizaki, G.
    • Journal Title

      J.Reprod.Dev. 52

      Pages: 685-693

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Visualization of primordial germ cells in vivo using GFP-nos1 3'UTR mRNA2006

    • Author(s)
      Saito, T., Fujimoto, T., Maegawa, S., Inoue, K., Tanaka, M., Arai, K., Yamaha, E.
    • Journal Title

      Int.J.Dev.Biol. 50

      Pages: 691-700

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Drastic mortality in tetraploid induction results from the elevation of ploidy in masu salmon Oncorhynchus masou.2006

    • Author(s)
      Sakao, S., Fujimoto, T., Kimura, S., Yamaha, E., Arai, K.
    • Journal Title

      Aquaculture. 252

      Pages: 147-160

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] ゼブラフィッシュの胚盤移植による生殖系列キメラの誘導.2005

    • Author(s)
      長井輝美
    • Journal Title

      水産学会誌 71

      Pages: 843-850

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Green fluorescent protein labeling of primordial germ cells using a nontransgenic method and its application for germ cell transplantation in Salmonidae.2005

    • Author(s)
      Yoshizaki, G.
    • Journal Title

      Biol. Reprod. 73

      Pages: 88-93

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Survival capacity of haploid-diploid goldfish chimeras.2005

    • Author(s)
      Tanaka, M., Yamaha, E., Arai, K.
    • Journal Title

      J.Exp.Zool. 301A

      Pages: 491-501

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] The formation of PGCs from the germline cells in spherical embryos derived from the blastodisc of 2-cell embryos in goldfish, Carassius auratus.2005

    • Author(s)
      Otani, S., Kitauchi, T., Sakao, S., Saito, T., Maegawa, S., Inoue, K., Arai, K., Yamaha, E.
    • Journal Title

      Int.J.Dev.Biol. 49

      Pages: 843-850

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Embryonic stages from cleavage to gastrula in the loach Misgurnus anguillicaudatus.2004

    • Author(s)
      Fujimoto, T.
    • Journal Title

      Zool. Sci. 21

      Pages: 747-755

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Requirement of localized maternal factors for zebrafish germ cell foramtion.2004

    • Author(s)
      Hashimoto, Y.
    • Journal Title

      Dev. Biol. 268

      Pages: 152-161

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Isolation of highly pure and viable primordial germ cells from rainbow trout by GFP-dependent flow cytometry.2004

    • Author(s)
      Kobayashi, T.
    • Journal Title

      Mol. Reprod. Dev. 67

      Pages: 91-100

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] E. Pluripotency of cryopreserved blastoderms of the goldfish.2004

    • Author(s)
      Kusuda, S.
    • Journal Title

      J. Exp. Zool. 301A

      Pages: 131-138

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] 胚盤移植により誘導されたゼブラフィッシュキメラ個体からの次世代.2004

    • Author(s)
      長井輝美
    • Journal Title

      水産育種 34

      Pages: 45-50

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] シマウキゴリの初期発生過程2004

    • Author(s)
      斎藤大樹
    • Journal Title

      SUISANZOUSYOKU 52

      Pages: 177-184

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The germ line lineage in ukigori, Gymnogobius species (Gobiidae : Teleostei) during embryonic development.2004

    • Author(s)
      Saito, T.
    • Journal Title

      Int. J. Dev. Biol. 48

      Pages: 1079-1085

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Surrogate broodstock produces salmonid.2004

    • Author(s)
      Takeuchi, Y.
    • Journal Title

      Nature 430

      Pages: 629-630

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Survival capacity of haploid-diploid goldfish chimeras.2004

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

      J. Exp. Zool. 301A

      Pages: 491-501

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Embryonic stages from cleavage to gastrula in the loach Misgurnus anguillicaudatus.2004

    • Author(s)
      Fujimoto, T., Kataoka, T., Saito, T., Otani, S., Yamaha, E., Arai, K.
    • Journal Title

      Zool.Sci. 21

      Pages: 747-755

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Requirement of localized maternal factors for zebrafish germ cell foramtion.2004

    • Author(s)
      Hashimoto, Y., Maegawa, S., Nagai, T., Yamaha, E., Suzuki, H., Yasuda, K., Inoue, K.
    • Journal Title

      Dev.Biol. 268

      Pages: 152-161

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Isolation of highly pure and viable primordial germ cells from rainbow trout by GFP-dependent flow cytometry.2004

    • Author(s)
      Kobayashi, T., Yoshizaki, G., Takeuchi, Y., Takeuchi, T.
    • Journal Title

      Mol.Reprod.Dev. 67

      Pages: 91-100

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Pluripotency of cryopreserved blastoderms of the goldfish.2004

    • Author(s)
      Kusuda, S., Teranishi, T., Koide, N., Nagai, T., Arai, K., Yamaha, E.
    • Journal Title

      J.Exp.Zool. 301A

      Pages: 131-138

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] The germ line lineage in ukigori, Gymnogobius species (Gobiidae : Teleostei) during embryonic development.2004

    • Author(s)
      Saito, T., Otani, S., Suzuki, T., Nakatsuji, T., Arai, K., Yamaha, E.
    • Journal Title

      Int.J.Dev.Biol. 48

      Pages: 1079-1085

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Surrogate broodstock produces salmonid.2004

    • Author(s)
      Takeuchi, Y., Yoshizaki, G., Takeuchi, T.
    • Journal Title

      Nature 430

      Pages: 629-630

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

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Published: 2008-05-27  

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