Browse > Article

Effect of Demecolcine-Assisted Enucleation and Recipient Cell Cycle Stage on the Development of Nuclear Transfer Bovine Embryos  

Back J. J. (College of Animal Resource Sciences, Kangwon National University)
Park C. K. (College of Animal Resource Sciences, Kangwon National University)
Yang B. K. (College of Animal Resource Sciences, Kangwon National University)
Kim C. I. (College of Animal Resource Sciences, Kangwon National University)
Cheong H. T. (College of Animal Resource Sciences, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.29, no.3, 2005 , pp. 175-180 More about this Journal
Abstract
This study was conducted to examine the effects of demecolcine-assisted enucleation and recipient cell cycle stage on the development of bovine somatic cell nuclear transfer (NT) embryos. In vitro cultured oocytes for $16\~20$ h were classified by first polar body (1st PB) extrusion and cell cycle stage (MI and MII) and treated $0.4\;{\mu}L/mL$ demecolcine for 40 min before enucleation. Enucleated oocytes were fused electrically with bovine ear skin cells, activated by Ca-ionophore+DMAP, and cultured in vitro. Most of eggs ($86.2\%$) treated with demecolcine protruded a chromosome mass and enucleated efficiently ($98.8\%$, (P<0.05). Demecolcine did not have a deteriorative effect on the development of NT embryos. Developmental rate of NT embryos reconstituted with oocytes extruded 1st PB significantly higher than that of NT embryos produced by oocytes without 1st PB ($18.2\%\;vs.\;4.6\%\cdot$, P<0.05). Cleavage and blastocyst formation rate of embryos reconstituted with MI oocytes ($69.4\%\;and\;5.9\%$, respectively) were significantly lower than those of embryos reconstituted with MII oocytes ($96.7\%\;and\;23.9\%$, respectively, P<0.05). From the present result, it is suggested that domecolcine is useful for the enucleation of recipient oocytes in bovine NT procedures, and MII oocytes rather than MI oocytes are more appropriate for recipient cytoplasm. Although, the potential to develop into blastocysts of NT embryos produced by 1st PB-nonextruded and MI oocytes was very low, these oorytes could be used for NT.
Keywords
Nuclear transfer; Enucleation; Demeco-lcine; Recipient cell cycle stage; In vitro development; bovine;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Cheong HT, Takahashi Y, Kanagawa H (1993): Birth of mice after transplantation of early cell-cycle-stage embryonic nuclei into enucleated oocytes. Biol Reprod 48:958-963   DOI   ScienceOn
2 Collas P, Robl JM (1991): Relationship between nuclear remodeling and development in nuclear transplant rabbit embryos. Biol Reprod 45:455-465   DOI   ScienceOn
3 Fulka J Jr, Moor RM (1993): Noninvasive chemical enucleation of mouse oocytes. Mol Reprod Dev 34: 427-430   DOI   ScienceOn
4 Tsunoda Y, Kato Y (2000): The recent progress on nuclear transfer in mammals. Zool Sci 17:1177-1184   DOI   ScienceOn
5 Zhang LS, Zhang KY, Yao LJ, Liu SZ, Yang CX, Zhong ZS, Zheng YL, Sun QY, Chen DY (2004): Somatic nucleus remodelling in immature and mature Rassir oocyte cytoplasm. Zygote 12:179-184   DOI   ScienceOn
6 Tani T, Kato Y, Tsunlda Y (2001): Direct exposure of chromosomes to nonactivated ovum cytoplasm is effective for bovine somatic cell nucleus reprogramming. Biol Reprod 64:324-330   DOI   ScienceOn
7 Yin XJ, Tani T, Yonemura I, Kawakami M, Miyamoto K, Hasegawa R, Kato Y, Tsunoda Y (2002a): Production of cloned pigs from adult somatic cells by chemically assisted removal of maternal chromosomes. Biol Reprod 67:442-446   DOI   ScienceOn
8 Gasparrini B, Gao S, Ainslie A, Fletcher J, McGarry M, Ritchie WA, Springbett AJ, Overstrom EW, Wilmut I, De Sousa P A (2003): Cloned mice derived from embryonic stem cell karyoplasts and activated cytoplasts prepared by induced enucleation. Biol Reprod 68: 1259-1266   DOI   ScienceOn
9 Ibanez E, Albertini DF, Overstrom EW (2003): Demecolcine-induced oocyte enucleation for somatic cell cloning; coordination between cell-cycle egress, kinetics of cortical cytoskeletal interactions, and second polar body extrusion. Biol Reprod 67:442-446   DOI   ScienceOn
10 Zernicka-Goetz M, Kubiak J, Antony C, Maro B (1993): Cytoskeletal organization of rat oocytes during metaphase II arrest and following abortive activation: A study by confocal laser scanning microscopy. Mol Reprod Dev 35:165-175   DOI   ScienceOn
11 Gao S, Gasparrini B, McGarry M, Ferrier T, Fletcher J, Harkness L (2002): Germinal vesicle material is essential for nucleus remodeling after nuclear transfer. BioI Reprod 67:928-934   DOI   ScienceOn
12 Elsheikh AS, Takahashi Y, Katagiri S, Kanagawa H (1997); Developmental ability of mouse late 2-cell stage blastomeres fused to chemically enucleated oocytes in vitro. J Vet Med Sci 59:107-113   DOI   ScienceOn
13 Liu JL, Sung LY, Barber M, Yang X (2002): Hypertonic medium treatment for localization of nuclear material in bovine metaphase II oocytes. Biol Reprod 67:1853-1863   DOI   ScienceOn
14 Cheong HT, Park TM, Ikeda K, Takahashi Y (2003): Cell cycle analysis of bovine cultured somatic cells by flow cytometry. Jpn J Vet Res 51:95-103
15 Smith LC (1993): Membrane and intracellular effects of ultraviolet irradiation with Hoechst 33342 on bovine secondary oocytes matured in vitro. J Reprod Fertil 99:39-44   DOI   ScienceOn
16 Mohamed Nour MS, Takahashi Y (1999): Preparation of young preactivated oocytes with high enucleation efficiency for bovine nuclear transfer. Theriogenology 51:661-666   DOI   ScienceOn
17 Polejaeva IA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, Dai Y, Boone J, Walke S, Ayares DL, Colman A, Campbell KHS (2000): Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407:505-509
18 Savard C, Novak S, Saint-Cyr A, Moreau M, Pothier F, Sirard MA (2004): Comparison of bulk enucleation methods for porcine oocytes. Mol Reprod Dev 67:70-76   DOI   ScienceOn
19 Yang X, Zhang L, Kovacs A, Tobback C, Foote RH (1990): Potential of hypertonic medium treatment for embryo micromanipulation: II. Assessment of nuclear transplantation methodology, isolation, subzona insertion and electrofusion of blastomeres to intact or functionally enucleated oocytes in rabbits. Mol Reprod Dev 27:118-129   DOI   ScienceOn
20 Choi JB, Kim CI, Park CK, Yang BK, Cheong HT (2004): Effect of activation time on the nuclear remodeling and in vitro development of nuclear transfer embryos derived from bovine somatic cells. Mol Reprod Dev 69:289-295   DOI   ScienceOn
21 Barnes F, Endebrock M, Looney C, Powell R, Westhusin M, Bondioli K (1993): Embryos cloning in cattle the use of in vitro matured oocytes. J Reprod Fertil 97:317-320   DOI   ScienceOn
22 Miyoshi K, Rzucidlo SJ, Gibbons JR, Arat S, Stice SL (2001): Development of porcine embryos reconstituted with somatic cells and enucleated metaphase I and II oocytes matured in a protein-free medium. BMC Dev Biol 1:12   DOI
23 Wehrend A, Meinecke B (2001): Kinetics of meiotic progression M-phase promoting factor(MPF) and mitogen-activated protein kinase(MAP kinase) activities during in vitro maturation of porcine and bovine oocytes: Species specific differences in the length of the meiotic stages. Animal Reprod Sci 66: 175-184   DOI   ScienceOn
24 Wilmut I, Schnieke AE, McWhirl J, Kind AJ, Campbell KHS (1997): Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813   DOI   ScienceOn
25 Kato Y, Tani T, Sotomaru Y, Kurokawa K, Kato J, Doguchi H, Yasue H, Tsunoda Y (1998): Eight calves cloned from somatic cells of a single adult. Science 282:2095-2098   DOI   PUBMED   ScienceOn
26 Yin XJ, Kato Y, Tsunoda Y (2002b): Effect of enucleation procedures and maturation conditions of the development of nuclear-transferred rabbit oocytes receiving male fibroblast cells. Reproduction 124: 41-47   DOI   PUBMED   ScienceOn
27 Campbell KHS, Loi P, Otaegui P, Wilmut I (1996a): Cell cycle co-ordination in embryo cloning by nuclear transfer. Rev Reprod 1:40-45   DOI   ScienceOn
28 Cheong HT, Takahashi Y, Kanagawa H (1994): Relationship between nuclear remodeling and subsequent development of mouse embryonic nuclei transferred to enucleated oocytes. Mol Reprod Dev 37:138-145   DOI   ScienceOn