Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer

Burgstaller, Joerg P.; Schinogl, Pamela; Dinnyés, András; Müller, Mathias; Steinborn, Ralf

doi:10.1186/1471-213x-7-141

Cited by 49 publications

(46 citation statements)

References 74 publications

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“…reconstructed embryos (Hiendleder et al 2003, Yang et al 2004, Burgstaller et al 2007. These organelles are involved in cellular metabolism with ATP production, apoptosis, regulation of calcium and cellular aging (Wang et al 2009b), so an inefficient nucleo-cytoplasmic communication for the regulation of mtDNA transcription and replication can lead to failure in embryonic development.…”

Section: Effect Of Iscnt and Aggregation On In Vitro Development Of Dmentioning

confidence: 99%

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Moro¹,

Hiriart²,

Buemo³

et al. 2015

Reproduction

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The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes were in vitro matured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P!0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.Reproduction (2015) 150 1-10

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Section: Effect Of Iscnt and Aggregation On In Vitro Development Of Dmentioning

confidence: 99%

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Moro¹,

Hiriart²,

Buemo³

et al. 2015

Reproduction

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show abstract

“…Although the rapid elimination of heteroplasmy facilitates the screening of homoplasmic mtDNA transformants, it hinders investigations on heteroplasmy dynamics, for example, determining the mechanism by which specific mitochondria with different mtDNAs are eliminated. Most naturally occurring heteroplasmic examples are small nucleotide polymorphisms or fragment length polymorphisms, which are difficult to identify (7,50).…”

Section: Discussionmentioning

confidence: 99%

“…The coexistence of different mtDNA molecules in a single eukaryotic cell, termed mtDNA heteroplasmy, can be observed from yeasts to higher organisms. Under natural conditions, mtDNA heteroplasmy arises from one of four sources: (i) partial damage to mtDNA from agents such as ionizing radiation and environmental toxins (17), (ii) mating of diploid fungal cells bearing different kinds of mtDNA (3,27), (iii) some types of sexual reproduction in higher organisms (7,53), and (iv) rare spontaneous heteroplasmy (50). Some plant and animal cells maintain a heteroplasmic state for extended periods (24,64).…”

Section: Discussionmentioning

confidence: 99%

Mitochondrial DNA Heteroplasmy in Candida glabrata after Mitochondrial Transformation

2010

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Genetic manipulation of mitochondrial DNA (mtDNA) is the most direct method for investigating mtDNA, but until now, this has been achieved only in the diploid yeast Saccharomyces cerevisiae. In this study, the ATP6 gene on mtDNA of the haploid yeast Candida glabrata (Torulopsis glabrata) was deleted by biolistic transformation of DNA fragments with a recoded ARG8 m mitochondrial genetic marker, flanked by homologous arms to the ATP6 gene. Transformants were identified by arginine prototrophy. However, in the transformants, the original mtDNA was not lost spontaneously, even under arginine selective pressure. Moreover, the mtDNA transformants selectively lost the transformed mtDNA under aerobic conditions. The mtDNA heteroplasmy in the transformants was characterized by PCR, quantitative PCR, and Southern blotting, showing that the heteroplasmy was relatively stable in the absence of arginine. Aerobic conditions facilitated the loss of the original mtDNA, and anaerobic conditions favored loss of the transformed mtDNA. Moreover, detailed investigations showed that increases in reactive oxygen species in mitochondria lacking ATP6, along with their equal cell division, played important roles in determining the dynamics of heteroplasmy. Based on our analysis of mtDNA heteroplasmy in C. glabrata, we were able to generate homoplasmic ⌬atp6 mtDNA strains.

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“…O DNA mitocondrial é herdado apenas por via materna, já que as mitocôndrias espermáticas que penetram no ooplasma durante a fertilização são destruídas (Hiendler et al, 2005). Na SCNT os embriões podem possuir mitocôndrias só procedentes do citoplasto (haloplasma) ou procedentes tanto da célula somática que doadora do núcleo quanto do citoplasto; ainda que a maioria dos animais estudados seja haloplasma, em raras ocasiões podem se encontrar um número elevado de mitocôndrias procedentes da célula somática (Burgstaller et al, 2007).…”

Section: O Genoma Mitocondrialunclassified

Clonagem de embriões bovinos a partir de células somáticas

2014

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Titulo en español: Clonación de embriones bovinos a partir de células somáticastitulo en ingles: Cloning bovine embryos from somatic cellsResumo: No presente artigo revisa-se dedicadamente a literatura do tema da clonagem tentando levar o leitor paulatinamente através dos avanços da técnica até alcançar as mais novas alternativas propostas pelos cientistas, e incluindo os erros mais comumente acontecidos nelas. A finalidade é mostrar o amplio panorama que tem aberto esta biotecnologia para a humanidade toda.Palavras chave: Transferência nuclear, fissão, paraclonagem, clonagem verdadeira, genoma mitocondrial.Resumen: En el presente artículo se revisa detenidamente la literatura en el tema de clonación, intentando llevar al lector paulatinamente a través de los avances de la técnica hasta alcanzar las más nuevas alternativas propuestas por los investigadores, incluyendo los errores más comúnmente cometidos. La finalidad es mostrar el amplio panorama que a abierto esta biotecnología para la humanidad.Palabras clave: Transferência nuclear, fissão, paraclonagem, clonagem verdadeira, genoma mitocondrial.Abstract: The present article provides a review of the pertinent literature on the topic of cloning. It aims to take the reader gradually through the advances made so far regarding the technique, to the newest alternatives proposed by researchers, including the most commonly committed errors, to unveil the broad panorama this biotechnology has opened up for humankind.Key words: Nuclear transfer, fission, paraclone colony, true cloning, mitochondrial genome.

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Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer

Cited by 49 publications

References 74 publications

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Mitochondrial DNA Heteroplasmy in Candida glabrata after Mitochondrial Transformation

Clonagem de embriões bovinos a partir de células somáticas

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