Ooplasm transfer and interspecies somatic cell nuclear transfer: heteroplasmy, pattern of mitochondrial migration and effect on embryo development

Sansinena, M.; Lynn, John W.; Bondioli, K. R.; Denniston, R.S.; Godke, R.A.

doi:10.1017/s0967199410000419

Cited by 20 publications

(21 citation statements)

References 41 publications

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“…Xenooplasmic transfer (XOT) in somatic cell models is a valuable tool to address nuclear and mtDNA interactions between different species (Kenyon & Moraes 1997, Trounce & Pinkert 2007. Unlike iSCNT embryos that usually arrest around the stage of EGA, XOT embryos develop beyond EGA (Sansinena et al 2011) and even into live offspring (Chiaratti et al 2010). Thus the XOT approach would provide a better model to address nuclear and cytoplasmic cross talk between different species.…”

Section: Introductionmentioning

confidence: 99%

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Amarnath

Choi²,

Moawad³

et al. 2011

Reproduction

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Inter-species somatic cell nuclear transfer (iSCNT) embryos usually fail to develop to the blastocyst stage and beyond due to incomplete reprogramming of donor cell. We evaluated whether using a karyoplast that would require less extensive reprogramming such as an embryonic blastomere or the meiotic spindle from metaphase II oocytes would provide additional insight into the development of iSCNT embryos. Our results showed that karyoplasts of embryonic or oocyte origin are no different from somatic cells; all iSCNT embryos, irrespective of karyoplast origin, were arrested during early development. We hypothesized that nuclear-cytoplasmic incompatibility could be another reason for failure of embryonic development from iSCNT. We used pig-mouse cytoplasmic hybrids as a model to address nuclear-cytoplasmic incompatibility in iSCNT embryos. Fertilized murine zygotes were reconstructed by fusing with porcine cytoplasts of varying cytoplasmic volumes (1/10 (small) and 1/5 (large) total volume of mouse zygote). The presence of pig cytoplasm significantly reduced the development of mouse zygotes to the blastocyst stage compared with control embryos at 120 h post-human chorionic gondotropin (41 vs 6 vs 94%, P!0.05; 1/10, 1/5, control respectively). While mitochondrial DNA copy numbers remained relatively unchanged, expression of several important genes namely Tfam, Polg, Polg2, Mfn2, Slc2a3 (Glut3), Slc2a1 (Glut1), Bcl2, Hspb1, Pou5f1 (Oct4), Nanog, Cdx2, Gata3, Tcfap2c, mt-Cox1 and mt-Cox2 was significantly reduced in cytoplasmic hybrids compared with control embryos. These results demonstrate that the presence of even a small amount of porcine cytoplasm is detrimental to murine embryo development and suggest that a range of factors are likely to contribute to the failure of inter-species nuclear transfer embryos.

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Section: Introductionmentioning

confidence: 99%

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Amarnath

Choi²,

Moawad³

et al. 2011

Reproduction

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“…The present study showed a good distribution of donor ooplasm containing mitochondria labelled with MitoTracker Green FM during early embryo development period (Figure ). In contrast, mitochondrial mismatch in interspecies OT between caprine and bovine showed early embryo developmental arrest as well as remained as a distinct cluster of transferred donor ooplasm into the developing embryo (Sansinena et al., ). These findings indicate that interspecies OT is unrestrainedly synchronized with recipient ooplasm but interspecies OT has difficulty in migrating throughout recipient ooplasm due to heteroplasmy or mitochondrial incompatibilities between different species.…”

Section: Discussionmentioning

confidence: 99%

Supplement of autologous ooplasm into porcine somatic cell nuclear transfer embryos does not alter embryo development

Lee

Jeon

et al. 2017

Reprod Domestic Animals

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Somatic cell nuclear transfer (SCNT) is considered as the technique in which a somatic cell is introduced into an enucleated oocyte to make a cloned animal. However, it is unavoidable to lose a small amount of the ooplasm during enucleation step during SCNT procedure. The present study was aimed to uncover whether the supplement of autologous ooplasm could ameliorate the oocyte competence so as to improve low efficiency of embryo development in porcine SCNT. Autologous ooplasm-transferred (AOT) embryos were generated by the supplementation with autologous ooplasm into SCNT embryos. They were comparatively evaluated with respect to embryo developmental potential, the number of apoptotic body formation and gene expression including embryonic lineage differentiation, apoptosis, epigenetics and mitochondrial activity in comparison with parthenogenetic, in vitro-fertilized (IVF) and SCNT embryos. Although AOT embryos showed perfect fusion of autologous donor ooplasm with recipient SCNT embryos, the supplement of autologous ooplasm could not ameliorate embryo developmental potential in regard to the rate of blastocyst formation, total cell number and the number of apoptotic body. Furthermore, overall gene expression of AOT embryos was presented with no significant alterations in comparison with that of SCNT embryos. Taken together, the results of AOT demonstrated inability to make relevant values improved from the level of SCNT embryos to their IVF counterparts.

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“…Nuclear-cytoplasmic incompatibility in iSCNT embryos was proved experimentally in goat-bovine (Sansinena, 2011) and pig-mouse (Amarnath et al, 2011) models. In contrast to injection of homologous ooplasm in bovine SCNT embryos that did not affect preimplantation embryo development, Interorder -NA Formation of nucleoli (Lagutina et al, 2011).…”

Section: Effect Of Heteroplasmy On Iscnt Embryo Developmentmentioning

confidence: 99%

Interspecies Somatic Cell Nuclear Transfer: Advancements and Problems

Lagutina

Fulka

Lazzari

et al. 2013

Cellular Reprogramming

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Embryologists working with livestock species were the pioneers in the field of reprogramming by somatic cell nuclear transfer (SCNT). Without the "Dolly experiment," the field of cellular reprogramming would have been slow and induced plutipotent cells (iPSCs) would not have been conceived. The major drive of the work in mammalian cloning was the interest of the breeding industry to propagate superior genotypes. Soon it was realized that the properties of oocytes could be used also to clone endangered mammalian species or to reprogram the genomes of unrelated species through what is known as interspecies (i) SCNT, using easily available oocytes of livestock species. iSCNT for cloning animals works only for species that can interbreed, and experiments with taxonomically distant species have not been successful in obtaining live births or deriving embryonic stem cell (ESC) lines to be used for regenerative medicine. There are controversial reports in the literature, but in most cases these experiments have underlined some of the cellular and molecular mechanisms that are incomplete during cell nucleus reprogramming, including the failure to organize nucleoli, silence somatic cell genes, activate the embryonic genome, and resume mitochondrial replication and function, thus indicating nucleus-cytoplasmic incompatibility.

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Ooplasm transfer and interspecies somatic cell nuclear transfer: heteroplasmy, pattern of mitochondrial migration and effect on embryo development

Cited by 20 publications

References 41 publications

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Supplement of autologous ooplasm into porcine somatic cell nuclear transfer embryos does not alter embryo development

Interspecies Somatic Cell Nuclear Transfer: Advancements and Problems

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