Analysis of 5-methyl cytosine and 5-hydromethyl cytosine content and distribution in different lineages during the bovine development
| Autor: | Hélène Jammes, Hélène Kiefer, Michel Guillomot, Audrey Prézelin, Anne Gabory, Juliette Salvaing, Renaud Fleurot, Evelyne Campion, Aladin Sassi, Nathalie Beaujean |
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| Přispěvatelé: | Biologie du Développement et Reproduction (BDR), Institut National de la Recherche Agronomique (INRA), 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), European Cooperation in Science and Technology (COST). BEL. |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: | |
| Zdroj: | Epigenetics and Periconception Environment 1. Annual Epiconcept Conference 1. Annual Epiconcept Conference, Apr 2013, Antalya, Turkey. pp.110 1. Annual Epiconcept Conference, European Cooperation in Science and Technology (COST). BEL., Apr 2013, Antalya, Turkey. pp.110 HAL |
| Popis: | DNA methylation of cytosines (5-mC) plays a crucial role in the epigenetic regulation of gene expression throughout development. The pattern of 5-mC is accurately established by de nova DNA methyltransferases (DNMT3a-3b) and inherited through mitosis by a maintenance DNA methyltransferase (DNMT1). Recently, another DNA modification, 5- hydroxymethylcytosine (5-hmC), was reported. 5-hmC is generated from 5-mC by TET proteins, and couId be an intermediate in demethylation pathways. Given the potential role of 5-hmC in nuclear reprogramming, it is crucial to obtain data in species with longer developmental lime courses than the mouse. This study aims to determine the tissue-, cell and nuclear content and distribution of 5-mC and 5-hmC in different bovine lineages during development. Bovine foetal (brain, liver, heart) and extraembryonic (allantois, amnion, chorion and cotyledon) tissues were collected at day 60 of gestation from 3 lemales and 3 males. Genomic DNA was prepared and digested by EcoRI, a methylation-non sensitive enzyme, to homogenize ils size. The 5-hmC or 5-mC content was measured using specifie 5-hmC or 5-mC immunoassays (Epigentek) and compared with mouse tissues (brain, muscle and liver from 3 females and 3 males at day 15 of gestation). ln arder to determine the distribution of 5-mC and 5-hmC in tissues/cells/nuclei, we performed immunostaining experiments with 5-mC and 5-hmC antibodies (Eurogentec and Active Motif). Image acquisitions were performed using the high-resolution Apotome microscope (Zeiss, MIMA2 facility, INRA, Jouy en Josas). Finally, the expression of the genes encoding DNMT3a, 3b, DNMT1 and TET was analysed in the different bovine tissues by quantitative PCR. Our data provide the first overview of the tissue/cell/nuclear distribution of 5-mC and 5-hmC during the development in bovine with a locus on the lineage origin, and will be a useful reference to determine the epigenetic alterations induced by situations affecting nuclear reprogramming, such as nuclear transfer |
| Databáze: | OpenAIRE |
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