• Title/Summary/Keyword: Transgenic cloned pig

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Ultrastructure in Porcine Oocytes following Intracytoplasmic Injection of Murine Spermatozoa

  • Kim, N. H.;Jun, S. H;Park, S. H.;J. Y. Yoon;D. I, Jin;S, H. Lee;Park, C. S.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.353-360
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    • 2002
  • Although successful pronuclear formation and apposition were seen in porcine oocytes following mouse sperm injection, little is known on the morphology of male and female pronuclei following sperm injection. The objective of this study is to describe the ultrastructure of porcine zygote following murine sperm injection in relation to the chronology of pronuclear S phase. At 40h ~ 44h following in vitro maturation, Cumulus cells were removed in TCM-HEPES with 0.1% hyaluronidase. Then, spermatozoa was injected into the cytoplasm of oocytes. After. injection, all oocytes were transferred to NCSU23 medium and cultured at 39$^{\circ}C$ under 5% $CO_2$ in air. Oocytes were fixed in 2% glutaraldehyde in Dulbeccos phosphate-buffered saline and observed by Transmission Electron Microscopy. Nuclear precursor bodies were observed in each pronucleus. A cluster of large and small granules was attached in the nucleolus precursor body. After the apposition of male and female chromatin, chromatin condensation was observed throughout the nucleoplasm and nucleolus precursor bodies and condensed chromatin in contact with clusters of small and large granules and the nuclear envelope were found in apposed pronuclear regions. These results suggest that non-species specific nuclear cytoplasmic interactions take place during pronuclear formation and apposition following sperm injection.

Onset of Pronuclear Formation and DNA Synthesis in Porcine Oocytes following Intracytoplasmic Injection of Porcine or Murine Spematozoa

  • Kim, N. H.;Cui, X. S;Kim, B. K .;S. H. Jun;D. I. Jin;Lee, S. H.;Park, C. S.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.361-368
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    • 2002
  • The onset of pronucleus formation and DNA synthesis in porcine oocytes following the injection of porcine or murine sperm was determined in order to obtain insights into species-specific paternal factors that contribute to fertilization. After 44h in vitro maturation, spermatozoa was injected into the cytoplasm of oocytes. After injection, all oocytes were transferred to NCSU23 medium and cultured at 39'E under 5% CO2 in air. Similar frequencies of oocytes with female pronuclei were observed after injection with porcine sperm or with murine sperm. In contrast, male pronuclei formed 8 to 9 h following the injection of porcine sperm, and 6 to 8 h following the injection of murine sperm. After pronucleus formation maternally derived microtubules were assembled and appeared to move both male and female pronuclei to the oocyte center. A few porcine oocytes entered metaphase 22 h after the injection of murine sperm, but normal cell division was not observed. The mean time of onset of S-phase in male pronuclei was 9.7 h following porcine sperm injection and 7.4 h following mouse sperm injection. These results suggested that DNA synthesis was delayed in both pronuclei until the sperm chromatin fully decondensed, and the sperm nuclear decondensing activity and microtubule nucleation abilities of the male centrosome are cell cycle dependent.

Umbilical Hernia and Repair in a Transgenic Male Cloned Pig

  • Kim, Geon A;Jin, Jun-Xue;Taweechaipaisankul, Anukul;Lee, Sanghoon;Kim, Min Jung;Lee, Byeong Chun
    • Journal of Veterinary Clinics
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    • v.35 no.5
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    • pp.226-228
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    • 2018
  • We generated a transgenic male cloned pig which was derived from fibroblast of white Yucatan miniature pig. After 2 weeks of birth, umbilical hernia which was not easily reduced was identified. Considering the usefulness of cloned pig, surgical treatment for umbilical hernia correction was performed and a cloned pig has been maintained healthy. This is the first report and can be useful for the treatments of umbilical hernia of cloned piglets.

Factors Influencing the Efficiency of In Vitro Embryo Production in the Pig

  • Lin, Tao;Lee, Jae Eun;Shin, Hyun Young;Oqani, Reza K.;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.39 no.2
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    • pp.29-36
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    • 2015
  • Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

Epigenetic characterization of the PBEF and TIMP-2 genes in the developing placentae of normal mice

  • Kim, Hong-Rye;Han, Rong-Xun;Diao, Yun-Fei;Park, Chang-Sik;Jin, Dong-Il
    • BMB Reports
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    • v.44 no.8
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    • pp.535-540
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    • 2011
  • Reprogramming errors, which appear frequently in cloned animals, are reflected by aberrant gene expression. We previously reported the aberrant expression of TIMP-2 and PBEF in cloned placenta and differential expression of PBEF genes during pregnancy. To examine the epigenetic modifications that regulate dynamic gene expression in developing placentae, we herein analyzed the mRNA and protein expression levels of PBEF and TIMP-2 in the placentae of normal mice during pregnancy and then examined potential correlations with epigenetic modifications. DNA methylation pattern analysis revealed no difference, but ChIP assays using antibodies against H3-K9/K14 and H4-K5 histone acetylation revealed that the H3-K9/K14 acetylation levels, but not the H4-K5 acetylation levels, of the TIMP-2 and PBEF loci were significantly correlated with their gene expression levels during placentation in normal mice. These results suggest that epigenetic changes may regulate gene expression level in the developing placentae of normal mice and that inappropriate epigenetic reprogramming might be one cause of the abnormal placentae seen in cloned animals.