Oocyte age and nuclear donor cell type affect the technical efficiency of somatic cloning in rabbits

Cervera, Rita P.; García-Ximénez, F.

doi:10.1017/s0967199403002181

Cited by 28 publications

(26 citation statements)

References 53 publications

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“…Our data do not confirm results from previous studies, in which cumulus cells were found to be better (Cervera & Garcia-Ximenez 2003) Figure 2 Histone acH3K9/14 status of in vivo fertilized and cloned embryos. Normal: in vivo fertilized embryos.…”

Section: Discussioncontrasting

confidence: 99%

Rabbit somatic cell cloning: effects of donor cell type, histone acetylation status and chimeric embryo complementation

Yang

Hao²,

Keßler³

et al. 2007

Reproduction

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The epigenetic status of a donor nucleus has an important effect on the developmental potential of embryos produced by somatic cell nuclear transfer (SCNT). In this study, we transferred cultured rabbit cumulus cells (RCC) and fetal fibroblasts (RFF) from genetically marked rabbits (Alicia/Basilea) into metaphase II oocytes and analyzed the levels of histone H3-lysine 9-lysine 14 acetylation (acH3K9/14) in donor cells and cloned embryos. We also assessed the correlation between the histone acetylation status of donor cells and cloned embryos and their developmental potential. To test whether alteration of the histone acetylation status affects development of cloned embryos, we treated donor cells with sodium butyrate (NaBu), a histone deacetylase inhibitor. Further, we tried to improve cloning efficiency by chimeric complementation of cloned embryos with blastomeres from in vivo fertilized or parthenogenetic embryos. The levels of acH3K9/14 were higher in RCCs than in RFFs (P!0.05). Although the type of donor cells did not affect development to blastocyst, after transfer into recipients, RCC cloned embryos induced a higher initial pregnancy rate as compared to RFF cloned embryos (40 vs 20%). However, almost all pregnancies with either type of cloned embryos were lost by the middle of gestation and only one fully developed, live RCC-derived rabbit was obtained. Treatment of RFFs with NaBu significantly increased the level of acH3K9/14 and the proportion of nuclear transfer embryos developing to blastocyst (49 vs 33% with non-treated RFF, P!0.05). The distribution of acH3K9/14 in either group of cloned embryos did not resemble that in in vivo fertilized embryos suggesting that reprogramming of this epigenetic mark is aberrant in cloned rabbit embryos and cannot be corrected by treatment of donor cells with NaBu. Aggregation of embryos cloned from NaBu-treated RFFs with blastomeres from in vivo derived embryos improved development to blastocyst, but no cloned offspring were obtained. Two live cloned rabbits were produced from this donor cell type only after aggregation of cloned embryos with a parthenogenetic blastomere. Our study demonstrates that the levels of histone acetylation in donor cells and cloned embryos correlate with their developmental potential and may be a useful epigenetic mark to predict efficiency of SCNT in rabbits.

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Section: Discussioncontrasting

confidence: 99%

Rabbit somatic cell cloning: effects of donor cell type, histone acetylation status and chimeric embryo complementation

Yang

Hao²,

Keßler³

et al. 2007

Reproduction

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“…Studies have shown that while nuclear maturation (emission of the first polar body) was mildly affected, the potential to form blastocysts in vitro was significantly compromised in oocytes with a poor cumulus expansion (Feng et al, 2007;Han et al, 2006). Furthermore, rates of cell fusion and embryonic development of NT embryos decreased significantly when aged oocytes were used for recipient cytoplasts (Cervera and García-Ximénez, 2003;Iwamoto et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Demecolcine-Assisted Enucleation of Goat Oocytes: Protocol Optimization, Mechanism Investigation, and Application to Improve the Developmental Potential of Cloned Embryos

Lan

Han

et al. 2008

Cloning and Stem Cells

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Although demecolcine-assisted enucleation has been performed successfully in porcine and cattle, the mechanism and protocol optimization of chemically assisted enucleation need further investigation. The present study optimized the protocol for goat oocyte enucleation and demonstrated that a 30-min treatment with 0.8 ng/mL demecolcine-induced cytoplasmic protrusions in over 90% of the oocytes. Rates of enucleation, cell fusion, and blastocyst formation were significantly higher after demecolcine-assisted than after blind aspiration enucleation, although differences in rates of live births remain to be unequivocally determined between the two treatments. The ability to form protrusions decreased significantly as spindles became less organized in aged oocytes and the oocytes with a poor cumulus expansion. More than 93% of the demecolcine-induced protrusions persisted for 2 h in the absence of cytochalasin B (CB) but most disappeared within 30 min of CB treatment. The spindle disintegrated, an actin-rich ring formed around the chromosome mass and the MAP kinase activity increased significantly after demecolcine treatment. When oocytes with induced protrusions were treated with CB, however, the contractile ring disappeared, the spindle reintegrated, and both MPF and MAP kinase activities decreased significantly. It is concluded that (1) cytoplasmic protrusions can be induced in goat oocytes with a very low concentration of demecolcine; (2) oocyte selection and enucleation can be achieved simultaneously with demecolcine treatment; and (3) an interactive effect between the MAP kinase, MPF, microfilaments and microtubules might be implicated in the control of cytoplasmic protrusion formation after demecolcine treatment.

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“…Many experimental designs in both research and clinical applications involve culture of matured oocytes prior to micromanipulation or insemination. For example, the cell cycle stage of the recipient cytoplasts must be finely controlled for the success of nuclear transfer for cloning (Zhou et al 2003, Wu et al 2007, and rates of cell fusion and embryonic development of nuclear transfer embryos decreased significantly when aged oocytes were used for recipient cytoplasts (Cervera & García-Ximénez 2003, Iwamoto et al 2005. Therefore, studies on the mechanisms and control of oocyte aging are important for the healthy reproduction of both humans and nonhuman mammals.…”

Section: Introductionmentioning

confidence: 99%

Pyruvate prevents aging of mouse oocytes

Liu

Wu²,

Lan³

et al. 2009

Reproduction

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Inhibiting oocyte aging is important not only for healthy reproduction but also for the success of assisted reproduction techniques. Although our previous studies showed that cumulus cells accelerated aging of mouse oocytes, the underlying mechanism is unknown. The objective of this paper was to study the effects of pyruvate and cumulus cells on mouse oocyte aging. Freshly ovulated mouse cumulus-oocyte complexes (COCs) or cumulus-denuded oocytes (DOs) were cultured in Chatot-Ziomek-Bavister (CZB) medium or COC-conditioned CZB medium supplemented with different concentrations of pyruvate before being examined for aging signs and developmental potential. Pyruvate supplementation to CZB medium decreased rates of ethanol-induced activation in both COCs and DOs by maintaining their maturation-promoting factor activities, but more pyruvate was needed for COCs than for DOs. Addition of pyruvate to the COC-conditioned CZB also alleviated aging of DOs. Observations on cortical granules, level of BCL2 proteins, histone acetylation, intracellular concentration of glutathione, and embryo development all confirmed that pyruvate supplementation inhibited aging of mouse oocytes. It is concluded that the aging of mouse oocytes, facilitated by culture in COCs, can be partially prevented by the addition of pyruvate to the culture medium.

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Oocyte age and nuclear donor cell type affect the technical efficiency of somatic cloning in rabbits

Cited by 28 publications

References 53 publications

Rabbit somatic cell cloning: effects of donor cell type, histone acetylation status and chimeric embryo complementation

Rabbit somatic cell cloning: effects of donor cell type, histone acetylation status and chimeric embryo complementation

Demecolcine-Assisted Enucleation of Goat Oocytes: Protocol Optimization, Mechanism Investigation, and Application to Improve the Developmental Potential of Cloned Embryos

Pyruvate prevents aging of mouse oocytes

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