1. Quantification of leukocyte genomic 5-methylcytosine levels reveals epigenetic plasticity in healthy adult cloned cattle
- Author
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Oliver J. Schmitz, Stefan Hiendleder, Jean-Paul Renard, Valeri Zakhartchenko, Béatrice de Montera, Dalia E. Zeihery, Eckhard Wolf, Gottfried Brem, Fabian Scheipl, Pascale Chavatte-Palmer, Horst-Dietrich Reichenbach, Sigrid Müller, Hélène Jammes, Biologie du développement et reproduction (BDR), Centre National de la Recherche Scientifique (CNRS)-École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), University of Wuppertal, Ludwig Maximilians University of Munich, University of Veterinary Sciences, Partenaires INRAE, Bavarian State Institute for Agriculture, University of Adelaide, European Commission FP6 contract No. FOOD-CT-2006-016250 ('SABRE'), Deutsche Forschungsgemeinschaft, Deutscher Akademischer Austausch Dienst (DAAD), French National Institute of Agronomical Research (INRA), and PhD grant from the French Ministry of Research (MRES)
- Subjects
Swine ,Cloning, Organism ,[SDV]Life Sciences [q-bio] ,Chemie ,Clone (cell biology) ,Monozygotic twin ,Biology ,somatic cell nuclear transfer ,Epigenesis, Genetic ,03 medical and health sciences ,0302 clinical medicine ,clones ,Transforming Growth Factor beta ,Genetic variation ,Genotype ,Leukocytes ,Animals ,Cell Lineage ,[INFO]Computer Science [cs] ,Epigenetics ,Oligonucleotide Array Sequence Analysis ,Skin ,030304 developmental biology ,Genetics ,0303 health sciences ,030219 obstetrics & reproductive medicine ,Stem Cells ,Original Articles ,Cell Biology ,Epigenome ,Immunohistochemistry ,Phenotype ,Gene Expression Regulation ,DNA methylation ,5-Methylcytosine ,Cell Division ,Signal Transduction ,Developmental Biology ,Biotechnology - Abstract
International audience; Successful somatic cell nuclear transfer (SCNT) requires epigenetic reprogramming of a differentiated donor cell nucleus. Incorrect reprogramming of epigenetic markings such as DNA methylation is associated with compromised prenatal development and postnatal abnormalities. Clones that survive into adulthood, in contrast, are assumed to possess a normalized epigenome corresponding to their normal phenotype. To address this point, we used capillary electrophoresis to measure 5-methylcytosine (5mC) levels in leukocyte DNA of 38 healthy female bovine clones that represented five genotypes from the Simmental breed and four genotypes from the Holstein breed. The estimated variance in 5mC level within clone genotypes of both breeds [0.104, 95% confidence interval (CI): 0.070–0.168] was higher than between clone genotypes (0, CI: 0–0.047). We quantified the contribution of SCNT to this unexpected variability by comparing the 19 Simmental clones with 12 female Simmental monozygotic twin pairs of similar age. In Simmental clones, the estimated variability within genotype (0.0636, CI: 0.0358–0.127) was clearly higher than in twin pairs (0.0091, CI: 0.0047–0.0229). In clones, variability within genotype (0.0636) was again higher than between genotypes (0, CI: 0–0.077). Twins, in contrast, showed lower variability within genotypes (0.0091) than between genotypes (0.0136, CI: 0.00250–0.0428). Importantly, the absolute deviations of 5mC values of individual SCNT clones from their genotype means were fivefold increased in comparison to twins. Further comparisons with noncloned controls revealed DNA hypermethylation in most of the clones. The clone-specific variability in DNA methylation and DNA hypermethylation clearly show that healthy adult SCNT clones must be considered as epigenome variants.
- Published
- 2010
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