DNA methylation in porcine preimplantation embryos developed in vivo and produced by in vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer

Deshmukh, Rahul S.; Østrup, Oľga; Østrup, Esben; Vejlsted, Morten; Niemann, Heiner; Lucas‐Hahn, Andrea; Petersen, Björn; Li, Juan; Callesen, Henrik; Hyttel, Poul

doi:10.4161/epi.6.2.13519

Cited by 100 publications

(78 citation statements)

References 57 publications

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“…What is more, there was no active demethylation in control zygotes produced from ovulated oocytes. Our observations of DNA methylation are in agreement with the work of Deshmukh et al (2011). Authors evaluated the methylation pattern of naturally produced zygotes with no demethylation of paternal genome observed.…”

Section: Discussionsupporting

confidence: 91%

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Dynamics of epigenetic remodeling in interspecies porcine zygotes

Barnetova¹,

Vacková²,

Firla³

2012

CZECH J ANIM SCI

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ABSTRACT:The process of active paternal chromatin demethylation after fertilization in the pig is not fully understood and very inconsistent data have been published by different research groups. We have applied the interspecies intracytoplasmic sperm injection (iICSI) to evaluate remodeling capabilities of porcine oocytes in more details. We injected mouse frozen-thawed sperm heads into porcine in vitro matured or ovulated oocytes, respectively. Embryos produced by intracytoplasmic sperm injection (ICSI) of boar spermatozoa into porcine ovulated oocytes (intraspecies) served as controls. Zygotes with 2-pronuclei were labeled with antibodies against certain epigenetic modifications (5-methylcytosine, 5-MeC; heterochromatin protein 1, HP1; trimethylation of H3/K4, H3/K4-me3; and dimethylation of H3/K9, H3/K9-me2). The labeling patterns were not different between zygotes produced from in vitro matured and ovulated oocytes. Both pronuclei were symmetrically labeled with 5-MeC, HP1, and H3/K4-me3 antibodies. Asymmetrical labeling was observed only with H3/K9-me2 antibody. The labeling of interspecies zygotes was similar to that of intraspecies zygotes. Moreover, the DNA demethylation was observed neither in control zygotes (intraspecies). The only difference observed between zygotes produced from in vitro matured and ovulated oocytes was in their ability to be activated. Intraspecies zygotes produced from ovulated oocytes were able to form the paternal pronucleus without additional activation; the zygotes produced from in vitro matured oocytes formed the paternal pronucleus only after additional activation with electric pulses. Our results show that the remodeling abilities of in vitro matured and ovulated oocytes are essentially similar. Moreover, it seems that reasons of inconsistent data reporting the active demethylation in the pig are more complicated and they are not associated exclusively with the oocyte quality.

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Section: Discussionsupporting

confidence: 91%

“…However, it seems that the active demethylation of paternal genome is absent also in high quality oocytes. As shown in the latest published paper, the active demethylation was not observed either in in vivo produced zygotes (Deshmukh et al, 2011).…”

Section: Dynamics Of Epigenetic Remodeling In Interspecies Porcine Zymentioning

confidence: 73%

Dynamics of epigenetic remodeling in interspecies porcine zygotes

Barnetova¹,

Vacková²,

Firla³

2012

CZECH J ANIM SCI

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“…Epigenetic reprogramming is crucial for the early development of the embryo, and the process is similar among various mammals like mouse, rat, pig, and cattle (Dean et al, 2001). In porcine preimplantation embryos, paternal pronuclei undergo active and rapid demethylation, whereas the maternal genome is passively demethylated during early cell cycles (Deshmukh et al, 2011). Subsequently cells undergo de novo remethylation during blastocyst formation and postimplantation development.…”

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confidence: 99%

Factors Determining the Efficiency of Porcine Somatic Cell Nuclear Transfer: Data Analysis with Over 200,000 Reconstructed Embryos

Liu

Dou

Xiang

et al. 2015

Cellular Reprogramming

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Data analysis in somatic cell nuclear transfer (SCNT) research is usually limited to several hundreds or thousands of reconstructed embryos. Here, we report mass results obtained with an established and consistent porcine SCNT system (handmade cloning [HMC]). During the experimental period, 228,230 reconstructed embryos and 82,969 blastocysts were produced. After being transferred into 656 recipients, 1070 piglets were obtained. First, the effects of different types of donor cells, including fetal fibroblasts (FFs), adult fibroblasts (AFs), adult preadipocytes (APs), and adult blood mesenchymal (BM) cells, were investigated on the further in vitro and in vivo development. Compared to adult donor cells (AFs, APs, BM cells, respectively), FF cells resulted in a lower blastocyst/reconstructed embryo rate (30.38% vs. 37.94%, 34.65%, and 34.87%, respectively), but a higher overall efficiency on the number of piglets born alive per total blastocysts transferred (1.50% vs. 0.86%, 1.03%, and 0.91%, respectively) and a lower rate of developmental abnormalities (10.87% vs. 56.57%, 24.39%, and 51.85%, respectively). Second, recloning was performed with cloned adult fibroblasts (CAFs) and cloned fetal fibroblasts (CFFs). When CAFs were used as the nuclear donor, fewer developmental abnormalities and higher overall efficiency were observed compared to AFs (56.57% vs. 28.13% and 0.86% vs.

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“…After the fertilization in mammalian zygotes (mouse, bovine, porcine, human), the next round of reprogramming begins. The paternal pronucleus is rapidly demethylated, while the maternal pronucleus undergoes passive demethylation (5). The demethylation process continues until the morula stage, when the cells located on the periphery of the embryo are predestined to become a trophoectoderm and the centrally located cells will form the ICM.…”

Section: Introductionmentioning

confidence: 99%

The Influence of Interspecies Somatic Cell Nuclear Transfer on Epigenetic Enzymes Transcription in Early Embryos

Morovic

Murín

Strejcek

et al. 2016

Macedonian Veterinary Review

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One of the main reason for the incorrect development of embryos derived from somatic cell nuclear transfer is caused by insufficient demethylation of injected somatic chromatin to a state comparable with an early embryonic nucleus. It is already known that the epigenetic enzymes transcription in oocytes and early embryos of several species including bovine and porcine zygotes is species-dependent process and the incomplete DNA methylation correlates with the nuclear transfer failure rate in mammals. In this study the transcription of DNA methyltransferase 1 and 3a (DNMT1, DNMT3a) genes in early embryonic stages of interspecies (bovine, porcine) nuclear transfer embryos (iSCNT) by RT-PCR were analyzed. Coming out from the diverse timing of embryonic genome activation (EGA) in porcine and bovine preimplantation embryos, the intense effect of ooplasm on transferred somatic cell nucleus was expected. In spite of the detection of ooplasmic DNA methyltransferases, the somatic genes for DNMT1 and DNMT3a enzymes were not expressed and the development of intergeneric embryos stopped at the 4-cell stage. Our results indicate that the epigenetic reprogramming during early mammalian development is strongly infl uenced by the ooplasmic environment.

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DNA methylation in porcine preimplantation embryos developed in vivo and produced by in vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer

Abstract: (2011) DNA methylation in porcine preimplantation embryos developed in vivo and produced by in vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer , Epigenetics, 6:2, 177-187,

Cited by 100 publications

References 57 publications

Dynamics of epigenetic remodeling in interspecies porcine zygotes

Dynamics of epigenetic remodeling in interspecies porcine zygotes

Factors Determining the Efficiency of Porcine Somatic Cell Nuclear Transfer: Data Analysis with Over 200,000 Reconstructed Embryos

The Influence of Interspecies Somatic Cell Nuclear Transfer on Epigenetic Enzymes Transcription in Early Embryos

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