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Comparison of Microtubule Distributions between Somatic Cell Nuclear Transfer and Parthenogenetic Porcine Embryos

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한국동물번식학회 (The Korean Society of Animal Reproduction)
초록

The aim of this study was to examine the microtubule distributions of somatic cell nuclear transfer (SCNT) and parthenogenetic porcine embryos. Porcine SCNT embryos were produced by fusion of serum-starved fetal fibroblast cells with enucleated oocytes. Reconstituted and mature oocytes were activated by electric pulses combined with 6-dimethlyaminopurine treatment. SCNT and parthenogenetic embryos were cultured in vitro for 6 days. Microtubule assembly of embryos was examined by confocal microscopy 1 hr and 20 hr after fusion or activation, respectively. The proportions of embryos developed to the blastocyst stage were 25.7% and 30.4% in SCNT and parthenogenetic embryos, respectively. The frequency of embryos showing β-tubulins was 81.8% in parthenogenetic embryos, whereas 31.3% in SCNT embryos 1 hr after activation or fusion. The frequency of the embryos underwent normal mitotic phase was low in SCNT embryos (40.6%) compared to that of parthenogenetic ones (59.7%) 20 hr after fusion or activation (p<0.05). The rate of SCNT embryos with an abnormal mitosis pattern is about twice compared to that of parthenogenetic ones. The spindle assembly and its distribution of SCNT embryos in the first mitotic phase were not different from those of parthenogenetic ones. The result shows that although microtubule distribution of porcine SCNT embryos shortly after fusion is different from parthenogenetic embryos, and the frequency of abnormal mitosis 20 hr after fusion or activation is slightly increased in SCNT embryos, microtubule distributions at the first mitotic phase are similar in both SCNT and parthenogenetic embryos.

목차
ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    In Vitro Maturation of Oocytes    Preparation of Porcine Fetal Fibroblasts    Micromanipulation    Electrofusion And Activation    Parthenogenesis and In Vitro Culture    Immunofluorescent Staining and Confocal Microscopy    Statistical Analysis   RESULTS    In Vitro Development of  Embryos    Microtubule Distnbution Shortly after Fusion or Activation    Microtubule Distribution in the First Mitotic Phase   DISCUSSION   REFERENCES
저자
  • Joo-Hee Park(School of Veterinary Medicine)
  • Dae-Jin Kwon(School of Veterinary Medicine)
  • Beom-Ki Lee(School of Veterinary Medicine)
  • In-Sun Hwang(College of Animal Life Science, Kangwon National University)
  • Choon-Keun Park(College of Animal Life Science, Kangwon National University)
  • Boo-Keun Yang(College of Animal Life Science, Kangwon National University)
  • Hee-Tae Cheong(School of Veterinary Medicine) Corresponding author