The analysis of chromatin remodeling and the staining for DNA methylation and histone acetylation do not provide definitive indicators of the developmental ability of inter-species cloned embryos

Lee, Eugine; Kim, Ji Hye; Park, Seon Mi; Jeong, Yeon Ik; Lee, Jong Yun; Park, Sun Woo; Choi, Jiho; Kim, Huen Suk; Kim, Sue; Hyun, Sang Hwan; Hwang, Woo Suk

doi:10.1016/j.anireprosci.2007.12.021

Cited by 14 publications

(12 citation statements)

References 25 publications

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“…Now it is well understood that the first step of any iSCNT experiments, such us remodeling of donor nucleus in the recipient ooplasm, is very conserved among species and depends on the maternally inherited factors of oocyte cytoplasm. The normal pattern of nucleus remodeling was followed as previously reported in different studies on mammalian nuclear transfer and iSCNT embryos (Arat et al, 2003;Dominko et al, 1999;Lee et al, 2008;Tarkowski and Balakier, 1980;Uhm et al, 2007). Thus, the biochemical mechanism of this process is universal and it works also in intraclass nuclear transfer embryos derived from chicken blastodermal cells and rabbit oocytes .…”

Section: Donor Nucleus Remodeling In Recipient Ooplasm Upon Nuclear Tmentioning

confidence: 53%

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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Section: Donor Nucleus Remodeling In Recipient Ooplasm Upon Nuclear Tmentioning

confidence: 53%

Interspecies Somatic Cell Nuclear Transfer: Advancements and Problems

Lagutina

Fulka

Lazzari

et al. 2013

Cellular Reprogramming

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“…Use of genetically modified donor cells for iSCNT not only offers a method of transgenic animal production but also provides solid evidence for donor cell origin of the resultant cloned embryos. The latter is particularly important because iSCNT embryos in several species are known to show normal chromatin remodeling and embryonic development beyond the embryonic genomic activation (EGA) stage, but they fail to develop further (Arat et al, 2003;Lee et al, 2008) due to aberrant nuclear reprogramming and gene expression (Arat et al, 2003;Chung et al, 2009;Lagutina et al, 2010;Lagutina et al, 2011;Ostrup et al, 2012;Wang et al, 2009;Wang et al, 2011). In this study, using EGFP-expressing donor cells, we have shown that pig oocytes are capable of supporting the early embryonic development of cattle, mice, and chicken nuclei at least up to the blastocyst stage.…”

Section: Discussionmentioning

confidence: 91%

“…Pig oocytes could reprogram somatic cells from rodents such as rabbit (Chen et al, 2006) and nonrodents such as dog (Lee et al, 2008;Sugimura et al, 2009), goat (Loi et al, 2011), tiger (Hashem et al, 2007), monkey (Qin et al, 2012), and cattle (Uhm et al, 2007). Pig oocytes were also shown to reprogram the somatic cells of rat that could not be reprogrammed by iSCNT into phylogenically closer mice oocytes (Sugawara et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Transgenic Chicken, Mice, Cattle, and Pig Embryos by Somatic Cell Nuclear Transfer into Pig Oocytes

Gupta

Das

Heo

et al. 2013

Cellular Reprogramming

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This study explored the possibility of producing transgenic cloned embryos by interspecies somatic cell nuclear transfer (iSCNT) of cattle, mice, and chicken donor cells into enucleated pig oocytes. Enhanced green florescent protein (EGFP)-expressing donor cells were used for the nuclear transfer. Results showed that the occurrence of first cleavage did not differ significantly when pig, cattle, mice, or chicken cells were used as donor nuclei (p>0.05). However, the rate of blastocyst formation was significantly higher in pig (14.9±2.1%; p<0.05) SCNT embryos than in cattle (6.3±2.5%), mice (4.2±1.4%), or chicken (5.1±2.4%) iSCNT embryos. The iSCNT embryos also contained a significantly less number of cells per blastocyst than those of SCNT pig embryos (p<0.05). All (100%) iSCNT embryos expressed the EGFP gene, as evidenced by the green florescence under ultraviolet (UV) illumination. Microinjection of purified mitochondria from cattle somatic cells into pig oocytes did not have any adverse effect on their postfertilization in vitro development and embryo quality (p>0.05). Moreover, NCSU23 medium, which was designed for in vitro culture of pig embryos, was able to support the in vitro development of cattle, mice, and chicken iSCNT embryos up to the blastocyst stage. Taken together, these data suggest that enucleated pig oocytes may be used as a universal cytoplast for production of transgenic cattle, mice, and chicken embryos by iSCNT. Furthermore, xenogenic transfer of mitochondria to the recipient cytoplast may not be the cause for poor embryonic development of cattle-pig iSCNT embryos.

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“…is used together when no DNA is observed and when the nuclear stage is not recognizable including picnotic DNA forming small intense patches~Otoi et al, 2002patches~Otoi et al, , 2004patches~Otoi et al, , 2006patches~Otoi et al, , 2007Luvoni et al, 2003;Rodrigues & Rodrigues, 2003;Willingham-Rocky et al, 2003;Rodrigues et al, 2004Rodrigues et al, , 2006Rodrigues et al, , 2009Vannucchi et al, 2006Vannucchi et al, , 2009Lee et al, 2007bLee et al, , 2008 Hossein et al, 2007;Hanna et al, 2008;Saikhun et al, 2008!. In some cases, "degenerated" oocytes are distinguished from those with unobservable DNA/undetermined nuclear stage~Saint-Dizier et al, 2004 with Hoechst and wide-field microscopy; Hatoya et al, 2009 with aceto orcein under phase contrast!.…”

Section: Discussionmentioning

confidence: 98%

Reliability of Hoechst 33342 Staining under Wide-Field Microscopy for Evaluation of the Nuclear Status of Living Dog Oocytes

et al. 2012

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Due to the marked cytoplasmic opacity of canine oocytes, the diagnosis of their nuclear status is difficult. The objective of the present study was to evaluate the accuracy of Hoechst staining observed under epifluorescence wide-field microscopy @living oocyte observation~LivOO!# by comparison to a reference technique @DNA staining with ethidium homodimer-2 under confocal microscopy; fixed oocyte observatioñ FixOO!#. Four Hoechst 33342 concentrations~200 ng, 500 ng, 1 mg, 2 mg/mL! were tested and 1 mg/mL was the lowest one with the lowest proportion of oocytes in which DNA was missed. At this concentration, LivOO procedure did not affect the degeneration rate. On 379 oocytes observed individually with the two techniques successively, diagnosis of meiosis resumption by LivOO was exact in 87.3% of the cases, but the meiosis resumption rate was underestimated~23.5% versus 34.3% with FixOO; p , 0.001!. Diagnosis for metaphase II was exact in 80% of the cases, but LivOO detected only 72.7% of the metaphase II oocytes present. Metaphase rates did not differ between LivOO and FixOO. This study contributes to a better interpretation of in vitro maturation results. The developmental potential of metaphase II canine oocytes sorted after Hoechst staining is to be evaluated.

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The analysis of chromatin remodeling and the staining for DNA methylation and histone acetylation do not provide definitive indicators of the developmental ability of inter-species cloned embryos

Cited by 14 publications

References 25 publications

Interspecies Somatic Cell Nuclear Transfer: Advancements and Problems

Interspecies Somatic Cell Nuclear Transfer: Advancements and Problems

Transgenic Chicken, Mice, Cattle, and Pig Embryos by Somatic Cell Nuclear Transfer into Pig Oocytes

Reliability of Hoechst 33342 Staining under Wide-Field Microscopy for Evaluation of the Nuclear Status of Living Dog Oocytes

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