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Effects of colchicine or demecolcine on cytoplasmic protrusions and assisted enucleation of golden hamster oocytes

Authors

  • Lingyan Wang,

    1. Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
    2. Department of Basic Veterinary Medicine, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
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    • These authors contributed equally to this work.

  • Han Jiang,

    1. Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
    2. Department of Basic Veterinary Medicine, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
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    • These authors contributed equally to this work.

  • Li Su,

    1. Second Clinical Hospital, Jilin University, Changchun, Jilin 130041, China
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  • Bo Tang,

    1. Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
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  • Dexue Li,

    1. Department of Basic Veterinary Medicine, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
    2. Academy of Military Medical Sciences, Beijing 100850, China
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  • Ziyi Li

    Corresponding author
    1. Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
    2. Department of Basic Veterinary Medicine, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China
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Jilin Province Key Laboratory of Animal Embryo Engineering, College of of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun, Jilin 130062, China. Tel./fax: +86 431 87836187. E-mail addresses: ziyili@jluhp.edu.cn

Abstract

To establish experimental protocols for cloning golden hamsters, optimal concentrations of colchicine and demecolcine were determined for inducing cytoplasmic protrusion (containing chromosomes) and assisting enucleation of their oocytes. Denuded oocytes at different ages were treated with 2.5–10 μg/ml of colchicine for 1–4 h or 0.02–0.6 μg/ml of demecolcine for 15–60 min. Cytoplasmic protrusions of oocytes were removed with a micromanipulation pipette. The results show that: 1) at 13.5–18 h post-hCG injection, ∼90% of oocytes treated for with 10 μg/ml of colchicine formed cytoplasmic protrusions, and in some oocytes enucleation occurred; 2) when treated with 0.4 μg/ml of demecolcine for 1 h, cytoplasmic protrusions 13.5–18 h post-hCG treatment were present in almost all oocytes; 3) after the protrusions induced by either treatment had been removed, the assisted enucleation rate was >80%, whereas it was ∼32% with blind enucleation.

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