Epigenetic modifications and related mRNA expression during bovine oocyte in vitro maturation
S. E. Racedo A B , C. Wrenzycki C , K. Lepikhov D , D. Salamone B , J. Walter D and H. Niemann A EA Institute of Farm Animal Genetics (FLI), Department of Biotechnology, Mariensee, 31535 Neustadt, Germany.
B Laboratorio de Biotecnología Animal, Facultad de Agronomía, Universidad de Buenos Aires, Av. San Martin 4453, C1417 Buenos Aires, Argentina.
C University of Veterinary Medicine Hannover, Clinic for Cattle, Reproductive Medicine Unit, Bischofsholer Damm 15, D-30173 Hannover, Germany.
D Department of Natural Sciences – Technical Faculty III, Biological Sciences, Genetics/Epigenetics, University of Saarland, 66123 Saarbrücken, Germany.
E Corresponding author. Email: heiner.niemann@fli.bund.de
Reproduction, Fertility and Development 21(6) 738-748 https://doi.org/10.1071/RD09039
Submitted: 4 March 2009 Accepted: 3 May 2009 Published: 1 July 2009
Abstract
The present study investigated the global pattern of two histone modifications and methylation of DNA during in vitro maturation of bovine oocytes retrieved from follicles of two different sizes (<2 mm and 2–8 mm). The methylation status of histone H3 at position lysine K9 (H3K9 me2), the acetylation status of histone H4 at position lysine K12 (H4K12ac) and the methylation of DNA were assessed by immunocytochemistry. In parallel, the relative abundance of mRNAs coding for proteins specifically involved in reprogramming, including HLA-B associated transcript 8 (G9A), suppressor of variegation 3-9 homolog 1 (SUV39H1), the somatic isoform of DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3b (DNMT3b) and zygote arrest 1 (ZAR1) was determined by RT-PCR. The α-H3K9 me2 signal was present in the GV stage and remained detectable until the end of the maturation period. α-H4K12ac antibody gave a stronger signal in GV and GVBD oocytes and markedly decreased after GVBD. The signal showing the methylation of DNA was present during the entire maturation period. The five transcripts showed a gene-specific expression profile. Results revealed the global patterns of H3K9 me2, H4K12ac, DNA methylation and the mRNA pool profiles of genes critically involved in epigenetic modifications during bovine oocyte maturation and their possible relationship with the acquisition of oocyte developmental competence and follicular development.
Additional keywords: developmental competence.
Acknowledgements
S.E.R. received a fellowship from the National Research Council from Argentina (CONICET). We are grateful to Thomas Jenuwein for providing the antibodies against H3K9 me2 and to Paul Hyttel’s lab for providing the antibodies against 5-methyl cytidine. The expert technical assistance of Brigitte Barg-Kues, Erika Lemme, Doris Herrmann and Khursheed Iqbal is gratefully acknowledged.
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