Quantitative analysis of mitochondrial DNA in porcine-mouse cloned embryos
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The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.
Abstract
The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.
참고문헌 (28)
- Fatira E, "Application of interspecific Somatic Cell Nuclear Transfer(iSCNT)in sturgeons and an unexpectedly produced gynogenetic sterlet with homozygous quadruple haploid", Sci Rep, 8, pp.5997~ (2018)[DOI]
- A. Cordova, "Choosing a culture medium for SCNT and iSCNT reconstructed embryos: from domestic to wildlife species", 한국축산학회지, 59(11), pp.1~14 (2017), 한국축산학회[DOI]
- Kono T, "Genomic imprinting is a barrier to parthenogenesis in mammals", Cytogenet Genome Res, 113, pp.31~35 (2006)[DOI]
- Dean W, "Epigenetic reprogramming in early mammalian development and following somatic nuclear transfer", Semin Cell Dev Biol, 14, pp.93~100 (2003)[DOI]
- Smith LC, "Genetic and epigenetic aspects of cloning and potential effects on offspring of cloned mammals", Cloning Stem Cells, 6, pp.126~132 (2004)[DOI]
- Jiang Y, "The fate of mitochondria in Ibexhirus reconstructed early embryos", Acta Biochim Biophys Sin, 36, pp.371~374 (2004)[DOI]
- Hua S, "Development of bovineovine interspecies cloned embryos and mitochondria segregation in blastomeres during preimplantation", Anim Reprod Sci, 105, pp.245~257 (2008)[DOI]
- Chen DY, "Interspecies implantation and mitochondria fate of panda-rabbit cloned embryos", Biol Reprod, 67, pp.637~642 (2002)[DOI]
- Ma LB, "Quantitative analysis of mitochondrial RNA in goat–sheep cloned embryos", Mol Reprod Dev, 75, pp.33~39 (2008)[DOI]
- Hashem MA, "Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger(Panthera tigris Altaica)", Mol Reprod Dev, 74, pp.403~411 (2007)[DOI]
- Chen T, "Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos", Mol Reprod Dev, 73, pp.313~317 (2006)[DOI]
- Naruse K, "Streptolysin-O treatment of fetal fibroblasts improves cell fusion and in vitro development of porcine nuclear transfer embryos", J Reprod Dev, 55, pp.236~239 (2009)[DOI]
- Ju-Yeon Kim, "Bezafibrate prevents aging in in vitro-matured porcine oocytes", 한국축산학회지, 63(4), pp.766~777 (2021), 한국축산학회[DOI]
- Naruse K, "Treatment of porcine oocytes with MEM vitamins during in vitro maturation improves subsequent blastocyst development following nuclear transfer", J Reprod Dev, 53, pp.679~684 (2007)[DOI]
- Rooney JP, "Mitochondrial regulation: methods and protocols", pp.23~38 (2015), Humana Press
- Ma H, "Incompatibility between nuclear and mitochondrial genomes contributes to an interspecies reproductive barrier", Cell Metab, 24, pp.283~294 (2016)[DOI]
- Jiang Y, "Interspecies somatic cell nuclear transfer is dependent on compatible mitochondrial DNA and reprogramming factors", PLOS ONE, 6, pp.e14805~ (2011)[DOI]
- Kim YS, "Development of cloned pig embryos by nuclear transfer following different activation treatments", Mol Reprod Dev, 70, pp.308~313 (2005)[DOI]
- Ikumi S, "Interspecies somatic cell nuclear transfer for in vitro production of Antarctic minke whale(Balaenoptera bonaerensis)embryos", Cloning Stem Cells, 6, pp.284~293 (2004)[DOI]
- Zhang T, "Effects of pyruvate on early embryonic development and zygotic genome activation in pigs", Theriogenology, 189, pp.77~85 (2022)[DOI]
- Amarnath D, "The novel use of modified pig zygotic medium for the efficient culture of the preimplantation mouse embryos", Theriogenology, 76, pp.1639~1646 (2011)[DOI]
- Im GS, "In vitro development of preimplantation porcine nuclear transfer embryos cultured in different media and gas atmospheres", Theriogenology, 61, pp.1125~1135 (2004)[DOI]
- Ma LB, "Development in vitro and mitochondrial fate of interspecies cloned embryos", Reprod Domest Anim, 43, pp.279~285 (2008)[DOI]
- Yang CX, "Quantitative analysis of mitochondrial DNAs in macaque embryos reprogrammed by rabbit oocytes", Reproduction, 127, pp.201~205 (2004)[DOI]
- Jiang Y, "In vitro culture and mtDNA fate of ibex–rabbit nuclear transfer embryos", Zygote, 13, pp.233~240 (2005)[DOI]
- Li Y, "Cloned endangered species takin(Budorcas taxicolor)by inter-species nuclear transfer and comparison of the blastocyst development with yak(Bos grunniens)and bovine", Mol Reprod Dev, 73, pp.189~195 (2006)[DOI]
- Meirelles FV, "Complete replacement of the mitochondrial genotype in a Bos indicus calf reconstructed by nuclear transfer to a Bos taurus oocyte", Genetics, 158, pp.351~356 (2001)[DOI]
- Steinborn R, "Coexistence of Bos taurus and B. indicus mitochondrial DNAs in nuclear transfer-derived somatic cattle clones", Genetics, 162, pp.823~829 (2002)[DOI]