Endometrial DNA methylation signatures during the time of breeding in relation to the pregnancy outcome in postpartum dairy cows fed a control diet or supplemented with rumen-protected methionine

Dessie Salilew-Wondim; Ernst Tholen; Eva Held-Hoelker; Karl Shellander; Carina Blaschka; Marc Drillich; Michael Iwersen; David Suess; Samuel Gebremedhn; Dawit Tesfaye; Claudia Parys; Ariane Helmbrecht; Jessie Guyader; Dennis Miskel; Nares Trakooljul; Klaus Wimmers; Michael Hoelker

doi:10.3389/fgene.2023.1267053

Endometrial DNA methylation signatures during the time of breeding in relation to the pregnancy outcome in postpartum dairy cows fed a control diet or supplemented with rumen-protected methionine

Front Genet. 2024 Jan 24:14:1267053. doi: 10.3389/fgene.2023.1267053. eCollection 2023.

Authors

Dessie Salilew-Wondim^{1

2}, Ernst Tholen², Eva Held-Hoelker², Karl Shellander¹, Carina Blaschka¹, Marc Drillich³, Michael Iwersen³, David Suess³, Samuel Gebremedhn⁴, Dawit Tesfaye⁴, Claudia Parys⁵, Ariane Helmbrecht⁵, Jessie Guyader⁵, Dennis Miskel², Nares Trakooljul⁶, Klaus Wimmers⁶, Michael Hoelker¹

Affiliations

¹ Department of Animal Science, Biotechnology and Reproduction of Farm Animals, University of Göttingen, Göttingen, Germany.
² Institute of Animal Sciences, Animal Breeding, University of Bonn, Bonn, Germany.
³ Clinical Unit for Herd Health Management, University Clinic for Ruminants, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine Vienna, Vienna, Austria.
⁴ Department of Biomedical Sciences, Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO, United States.
⁵ Evonik Operations GmbH, Hanau, Germany.
⁶ Research Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

Abstract

Post calving metabolic stress reduces the fertility of high producing dairy cows possibly by altering the expression of genes in the maternal environment via epigenetic modifications. Therefore, this study was conducted to identify endometrial DNA methylation marks that can be associated with pregnancy outcomes in postpartum cows at the time of breeding. For this, twelve days post-calving, cows were either offered a control diet or supplemented daily with rumen-protected methionine. Cows showing heat 50-64 days postpartum were artificially inseminated. Endometrial cytobrush samples were collected 4-8 h after artificial insemination and classified based on the pregnancy out comes as those derived from cows that resulted in pregnancy or resulted in no pregnancy. The DNAs isolated from endometrial samples were then subject to reduced representative bisulfite sequencing for DNA methylation analysis. Results showed that in the control diet group, 1,958 differentially methylated CpG sites (DMCGs) were identified between cows that resulted in pregnancy and those that resulted in no pregnancy of which 890 DMCGs were located on chr 27: 6217254-6225600 bp. A total of 537 DMCGs were overlapped with 313 annotated genes that were involved in various pathways including signal transduction, signalling by GPCR, aldosterone synthesis and secretion. Likewise, in methionine supplemented group, 3,430 CpG sites were differentially methylated between the two cow groups of which 18.7% were located on Chr27: 6217254-6225600 bp. A total of 1,781 DMCGS were overlapped with 890 genes which involved in developmental and signalling related pathways including WNT-signalling, focal adhesion and ECM receptor interaction. Interestingly, 149 genes involved in signal transduction, axon guidance and non-integrin membrane-ECM interactions were differentially methylated between the two cow groups irrespective of their feeding regime, while 453 genes involved in axon guidance, notch signalling and collagen formation were differentially methylated between cows that received rumen protected methionine and control diet irrespective of their fertility status. Overall, this study indicated that postpartum cows that could potentially become pregnant could be distinguishable based on their endometrial DNA methylation patterns at the time of breeding.

Keywords: endometrium; methionine; methylation; postpartum; pregnancy.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was funded by Evonik Operations GmbH. The funding body has no role in designing the experiment, collection of data, analysis and interpretation of data, writing the manuscript, or the decision to submit it for publication.