Developmental Enhancement of Intracytoplasmic Sperm Injection (ICSI)-Generated Quail Embryos by Phospholipase Cζ cRNA

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著者

    • ATSUMI Yusuke
    • Department of Food Production Science, Faculty of Agriculture, Shinshu University
    • TSUKADA Akira
    • Laboratory of Animal Physiology, Graduate School of Bioagricultural Sciences, Nagoya University
    • SAITO Noboru
    • Laboratory of Animal Physiology, Graduate School of Bioagricultural Sciences, Nagoya University
    • SHIMADA Kiyoshi
    • Laboratory of Animal Physiology, Graduate School of Bioagricultural Sciences, Nagoya University

抄録

The present study was conducted to improve the rate and stage of blastoderm and embryo development of quail oocytes by intracytoplasmic sperm injection (ICSI) with quail phospholipase Cζ (PLCζ) cRNA. Blastoderm development of quail oocytes cultured for 24 or 72h were staged under a stereomicroscope. The blastoderms were stained with 4,6-diamidino-2-phenylindole (DAPI) for cell division progress. A quail PLCζ cDNA clone was isolated from sperm by RT-PCR. The 1978bp nucleotide sequence contained an ORF encoding 637 amino acids of protein. The deduced quail PLCζ protein consists of EF-hand domain, X and Y catalytic domain and C2 domain, but lacked a plextrin-homology (PH) domain at the N-terminus. RT-PCR analysis revealed that PLCζ mRNA is expressed only in the testis. ICSI into a quail oocyte without PLCζ cRNA induced blastodermal development in 16% (4/25) of the oocytes which reached stages III-VI 24h after culture. In contrast, ICSI together with PLCζ cRNA (in 3n<I>l</I> at 60μg/m<I>l</I>) into a quail oocyte induced blastoderm development more than 2 fold (36.8%, 7/19) with embryos reaching stages VI-VII. Furthermore, 72h after culture ICSI into a quail oocyte without PLCζ cRNA induced embryo development in 15.4% (2/13) of the oocytes and reached stages V-VII. In contrast, ICSI together with PLCζ cRNA (in 3n<I>l</I> at 60μg/m<I>l</I>) into a quail oocyte induced 3 fold more embryo development (50%, 9/18) with embryos between stages VI and over X. The microinjection of PLCζ cRNA significantly improved the conventional ICSI method by enhancing the proportion and the stages of embryo development. Accordingly, the present PLCζ cRNA microinjection method in birds may contribute to assist in the production of transgenic birds and protection of endangered species of birds.

収録刊行物

  • The journal of poultry science  

    The journal of poultry science 45(2), 152-158, 2008-04-25 

    日本家禽学会

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各種コード

  • NII論文ID(NAID)
    10021220283
  • NII書誌ID(NCID)
    AA11564513
  • 本文言語コード
    ENG
  • 資料種別
    NOT
  • ISSN
    13467395
  • NDL 記事登録ID
    9469303
  • NDL 雑誌分類
    ZR22(科学技術--農林水産--畜産)
  • NDL 請求記号
    Z54-J694
  • データ提供元
    CJP書誌  CJP引用  NDL  J-STAGE 
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