Effects of Maturational Age of Porcine Oocytes on the Induction of Activation and Development In Vitro Following Somatic Cell Nuclear Transfer.

Ikeda, Kazuhiko; Takahashi, Yoshiyuki

doi:10.1292/jvms.63.1003

Cited by 32 publications

(33 citation statements)

References 33 publications

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“…MPF activity in cytoplasts plays a key role(s) for development of NT embryos, since the length of the maturation period affects the ability of porcine cytoplasts to support the development of NT embryos (Cheong et al 2000, Ikeda & Takahashi 2001, Miyoshi et al 2002. In this study, we found that high MPF activity elevated in cytoplasts improved the development of NT embryos.…”

Section: Mpf Activity In Porcine Nt Embryossupporting

confidence: 54%

“…When the donor nuclei in G0/G1 of the cell cycle are transferred into recipient cytoplasts with a high level of maturation promoting factor (MPF) before activation, they are transformed to nuclear envelope breakdown (NEBD) and then premature chromosome condensation (PCC) (Ikeda & Takahashi 2001). In the mouse and bovine, this PCC of the transferred nucleus was related to the success of cloning (Shin et al 2002, Wakayama et al 1998).…”

Section: Introductionmentioning

confidence: 99%

“…When oocytes with low MPF activity are fertilized with sperm, they show abnormal pronuclear formation and embryo development, indicating that MPF activity is thought as an indicator to evaluate the quality of the oocyte (Kikuchi et al 1995). However, MPF activity in recipient cytoplasts and NT embryos has remained poorly understood, although the maturational cultivation period of the oocytes used as recipient cytoplasts can affect the developmental ability of NT embryos (Cheong et al 2000, Ikeda & Takahashi 2001, Miyoshi et al 2002. Furthermore, the relationship between morphological remodeling such as NEBD or PCC and the development after activation has been poorly understood (Uhm et al 2000).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer

Kawahara¹,

Wakai²,

Yamanaka³

et al. 2005

Reproduction

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When the nucleus in G0/G1 phase is transferred to an enucleated oocyte by nuclear transfer (NT), its nuclear envelope is broken, followed by condensation of chromosome structure by maturation promoting factor (MPF). This morphological remodeling of the transferred interphase nucleus seems to be essential for subsequent development of NT embryos. In this study, we treated porcine NT embryos with caffeine, which has been reported to increase MPF activity, to keep their MPF level high during NT. When 2.5 mM caffeine was added to the handling medium, the proportion of NT embryos showing condensed chromosome increased significantly (P < 0.05). In NT embryos treated with caffeine, the activity of p34 cdc2 kinase was significantly (P < 0.05) higher than in those without caffeine at 3 h post-injection. In addition, the rate of development to the blastocyst stage after activation was significantly (P < 0.05) higher in NT embryos treated with caffeine. These results indicate that caffeine treatment can increase not only the rate of chromosome condensation but also the developmental rate to the blastocyst stage of porcine NT embryos. This action is most likely due to the support/increase of MPF activity throughout the process of NT.Reproduction (

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Section: Mpf Activity In Porcine Nt Embryossupporting

confidence: 54%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer

Kawahara¹,

Wakai²,

Yamanaka³

et al. 2005

Reproduction

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“…Maturational age of porcine oocytes affects the potential of subsequent embryo development. In pigs, newly matured oocytes showed lower developmental competence after IVF [34] or parthenogenetic activation [35] compared with oocytes cultured for an additional 8-10 h. This might be due to inadequate cytoplasmic maturation and/or immaturity of the metaphase spindle of MII stage oocytes immediately after MIIarrest [36]. The time at which the oocytes reached the MII stage in mNCSU37+PVA was delayed compared with the other maturation media (Fig.…”

Section: Discussionmentioning

confidence: 93%

Defined System for In Vitro Production of Porcine Embryos Using a Single Basic Medium

2008

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Abstract. We have previously indicated that porcine blastocysts can be produced by in vitro fertilization (IVF) and culture (IVC) in chemically defined porcine gamete medium (PGM) and porcine zygote medium (PZM)-5, respectively, In the present study, the effects of basic media and macromolecular components on in vitro maturation (IVM) were investigated to develop a defined system for in vitro embryo production using a single basic medium through IVM, IVF and IVC. Porcine immature oocytes were matured in porcine oocyte medium (POM) or modified North Carolina State University (mNCSU) 37, which were supplemented with either 10% (v/v) porcine follicular fluid (pFF) or 3 mg/ml polyvinyl alcohol (PVA) as a macromolecular component (designated POM+pFF, POM+PVA, mNCSU37+pFF and mNCSU37+PVA). In the maturation with mNCSU37+PVA, the percentages of oocytes that reached the metaphase II stages were significantly lower than those in the other treatments. Following IVM with the above media, oocytes were treated with an electrical stimulus and cycloheximide for parthenogenetic activation and were cultured in PZM-5 for 5 days. The rates of cleavage and blastocyst formation of parthenogenetic oocytes were significantly lowered for maturation with mNCSU37+PVA compared with the other treatments, while there were no significant differences in the total numbers of cells in blastocysts among the treatments. Following IVF and IVC, the rates of penetration, male pronucleus formation, cleavage and blastocyst formation were significantly lower when oocytes were matured in mNCSU37+PVA than in other maturation media. The normal fertilization rate was significantly higher in POM+PVA compared with the other treatments, although the total number of cells in blastocysts was reduced with the addition of PVA to both POM and mNCSU37 compared with pFF supplementation. These results demonstrate that porcine blastocysts can be produced by the defined system using a single basic medium. Key words: Defined system, In vitro production (IVP), Maturation, Porcine embryo, Porcine zygote medium (PZM) (J. Reprod. Dev. 54: [208][209][210][211][212][213] 2008) he establishment of an in vitro production (IVP) system for preimplantation embryos that can develop to full term after transfer will contribute to a better understanding of the physiology of embryonic development in early pregnancy and the control of animal reproduction, including embryo transfer, transgenesis and cloning [1]. Numerous studies have investigated the ability of porcine oocytes and embryos to develop in vitro using a wide variety of culture media [2][3][4][5][6][7]. Recently, we developed a chemically defined medium (porcine zygote medium: PZM) for in vitro culture (IVC) of porcine zygotes based on the composition of pig oviduct fluid [8]. Moreover, we established a chemically defined system for in vitro fertilization (IVF) of porcine in vitro-matured oocytes using porcine gamete medium (PGM), modified from PZM, as a basic medium [9], and the in vivo viability of blastocysts produced i...

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“…In both species, when a donor nucleus in the G0/G1 stage of the cell cycle is transferred to an enucleated oocyte, the PCC of the transferred nucleus is related to the success of cloning and probably reprogramming [12]. Donor nuclei in the G0/G1 stage show nuclear envelope breakdown (NEBD) and PCC when they are transferred into recipient cytoplasts with high levels of maturation/ metaphase promoting factor (MPF) [19] before activation [20]. Methods that induce PCC in SCNT oocytes result in increased developmental potential, indicating that these processes may promote reprogramming of gene expression in the donor nucleus [12,[21][22][23][24][25][26].…”

Section: Discussionmentioning

confidence: 99%

Spindle Formation and Microtubule Organization During First Division in Reconstructed Rat Embryos Produced by Somatic Cell Nuclear Transfer

Tomioka

Mizutani

Yoshida

et al. 2007

Journal of Reproduction and Development

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Abstract. The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 µM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-α-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization.

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Effects of Maturational Age of Porcine Oocytes on the Induction of Activation and Development In Vitro Following Somatic Cell Nuclear Transfer.

Cited by 32 publications

References 33 publications

Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer

Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer

Defined System for In Vitro Production of Porcine Embryos Using a Single Basic Medium

Spindle Formation and Microtubule Organization During First Division in Reconstructed Rat Embryos Produced by Somatic Cell Nuclear Transfer

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