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

Park, Joo-Hee (School of Veterinary Medicine, Kangwon National University)
Kwon, Dae-JinK (School of Veterinary Medicine, Kangwon National University)
Lee, Beom-Ki (School of Veterinary Medicine, Kangwon National University)
Hwang, In-Sun (College of Animal Life Science, Kangwon National University)
Park, Choon-Keun (College of Animal Life Science, Kangwon National University)
Yang, Boo-Keun (College of Animal Life Science, Kangwon National University)
Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University)
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Abstract
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 $\beta$-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.
Keywords
Somatic cell nuclear transfer; Microtubule distribution; Nuclear progression; Confocal microscopy; Porcine embryos;
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1 Collas P, Robl JM (1991): Relationship between nuclear remodeling and development in nuclear transplant rabbit embryos. Biol Reprod 45:455-465   DOI   ScienceOn
2 Kwon DJ, Park CK, Yang BK, Kim CI, Cheong HT (2007): Effect of maturational age of recipient oocytes and activation conditions on the development of porcine fetal fibroblast nuclear transfer embryos. Anim Reprod Sci 100:211-215   DOI   ScienceOn
3 Petters RM, Wells KD (1993): Culture of pig embryos. J Reprod Fertil (Suppl) 48:61-73   PUBMED
4 Thomson EM, Legouy E, Renard JP (1998): Mouse embryos do not wait for the MBT: chromatin and RNA polymerase remodeling in genome activation at the onset of development. Dev Genet 22:31-41   DOI   ScienceOn
5 Collas P, Pinto-Correia C, Ponce De Leon FA, Robl JM (1992): Development of nuclear transplant rabbit embryos: Influence of the cell cycle stage of the donor nucleus. Biol Reprod 46:492-550   DOI   ScienceOn
6 Dai Y, Wang L, Wang H, Liu Y, Li N, Lyu Q, Keefe DL, Albertini DF, Liu L (2006): Fate of centrosomes following somatic cell nuclear transfer (SCNT) in bovine oocytes. Reproduction 131:1051-1061   DOI   PUBMED   ScienceOn
7 Kim NH, Funahashi H, Prather RS, Schatten G, Day BN (1996): Microtubule and microfilament dynamics in porcine oocytes during meiotic maturation. Mol Reprod Dev 43:248-255   DOI   ScienceOn
8 Zhong ZS, Zhang G, Meng XQ, Zhanga YL, Chena DY, Schattenc H, Sun QY (2005): Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos. Exp Cell Res 306:35-46   DOI   ScienceOn
9 Nguyen VT, Wakayama S, Kishigami S, Wakayama T (2004): Spindle morphogenesis and the morphology chromosomes in mouse nuclear transfer: an abnormal start in cloning of mice. Reprod Fertil Dev 16:153–153   ScienceOn
10 Yoshioka K, Suzuki C, Tanaka I, Anas IMK, Iwamura S (2002): Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol Reprod 66:112-119   DOI   ScienceOn
11 Wilmut I, Schnieke AE, McWhir J, Kind AL, Campbell KHS (1997): Viable offspring derived from fetal and adult mammalian cells. Nature 385:910-813   DOI   ScienceOn
12 Pinto-Correia C, Collas P, Ponce De Leon FA, Robl JM (1993): Chromatin and microtubule organization in the first cell cycle in rabbit parthenotes and nuclear transplant embryos. Mol Reprod Dev 34:33-42   DOI   ScienceOn
13 Navara CS, First NL, Schatten G (1994) Microtubule organization in the cow during fertilization, polyspermy, parthenogenesis, and nuclear transfer: The role of the sperm aster. Dev Biol 162:29-40   DOI   ScienceOn
14 Campbell KHS (1999): Nuclear transfer in farm animal species. Cell Dev Biol 10:245-252   DOI   PUBMED   ScienceOn
15 Shin MR, Park SW, Shim H, Kim NH (2002): Nuclear and microtubule reorganization in nuclear-transferred bovine embryos. Mol Reprod Dev 62:74- 82   DOI   ScienceOn
16 Zhong Z, Spate L, Hao Y, Li R, Lai L, Katayama M, Sun QY, Prather RS, Schatten H (2007); Remodeling of centrosomes in intraspecies and interspecies nuclear transfer porcine embryos. Cell Cycle 6:1510- 1521   PUBMED   ScienceOn
17 Niemann H, Reichelt B (1993): Manipulating early pig embryos. J Reprod Fertil (Suppl) 48:75-94   PUBMED
18 Tomioka I, Mizutani E, Yoshida T, Sugawara A, Inai K, Sasada H, Sato E (2007): Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer. J Reprod Dev 53:835-842   DOI   ScienceOn
19 Kirshner M, Mitchison T (1986): Beyond self-assembly: from microtubules to morphogenesis. Cell 45: 329-342   DOI   ScienceOn