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Acetylation and methylation profiles of H3K27 in porcine embryos cultured in vitro

Published online by Cambridge University Press:  11 July 2017

Luciana Simões Rafagnin Marinho
Affiliation:
Laboratory of Animal Reproduction, State University of Londrina (UEL), Rodovia Celso Garcia Cid, Km 380, s/n – Campus Universitário, CEP 86057–970, Londrina, PR, Brazil
Vitor Braga Rissi
Affiliation:
Laboratory of Biotechnology and Animal Reproduction – BioRep, Veterinary Hospital, Federal University of Santa Maria, Av. Roraima, 1000 – Camobi, CEP 97105–900, Santa Maria, RS, Brazil
Andressa Guidugli Lindquist
Affiliation:
Laboratory of Animal Reproduction, State University of Londrina (UEL), Rodovia Celso Garcia Cid, Km 380, s/n – Campus Universitário, CEP 86057–970, Londrina, PR, Brazil
Marcelo Marcondes Seneda*
Affiliation:
Laboratory of Animal Reproduction, State University of Londrina (UEL), Rodovia Celso Garcia Cid, Km 380, s/n – Campus Universitário, CEP 86057–970, Londrina, PR, Brazil.
Vilceu Bordignon
Affiliation:
Department of Animal Science, McGill University, 21111 Lakeshore Road, Sainte Anne de Bellevue, Quebec H9X 3V9, Canada
*
All correspondence to: Marcelo Marcondes Seneda. Laboratory of Animal Reproduction, State University of Londrina (UEL), Rodovia Celso Garcia Cid, Km 380, s/n – Campus Universitário, CEP 86057–970, Londrina, PR, Brazil. E-mail: [email protected]

Summary

Methylation and acetylation of histone H3 at lysine 27 (H3K27) regulate chromatin structure and gene expression during early embryo development. While H3K27 acetylation (H3K27ac) is associated with active gene expression, H3K27 methylation (H3K27me) is linked to transcriptional repression. The aim of this study was to assess the profile of H3K27 acetylation and methylation (mono-, di- and trimethyl) during oocyte maturation and early development in vitro of porcine embryos. Oocytes/embryos were fixed at different developmental stages from germinal vesicle to day 8 blastocysts and submitted to an immunocytochemistry protocol to identify the presence and quantify the immunofluorescence intensity of H3K27ac, H3K27me1, H3K27me2 and H3K27me3. A strong fluorescent signal for H3K27ac was observed in all developmental stages. H3K27me1 and H3K27me2 were detected in oocytes, but the fluorescent signal decreased through the cleavage stages and rose again at the blastocyst stage. H3K27me3 was detected in oocytes, in only one pronucleus in zygotes, cleaved-stage embryos and blastocysts. The nuclear fluorescence signal for H3K27me3 increased from the 2-cell stage to 4-cell stage embryos, decreased at the 8-cell and morula stages and increased again in blastocysts. Different patterns of the H3K27me3 mark were observed at the blastocyst stage. Our results suggest that changes in the H3K27 methylation status regulate early porcine embryo development as previously shown in other species.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2017 

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