Abstract
Introduction
Bovine female and male embryos differentially release metabolites with signalling effects to culture media. However, it is unknown if the embryo-maternal interface (EMI) metabolome is modified by embryonic sex.
Objective
To analyse using a combination of 1H NMR and a co-culture of endometrial cells the EMI.
Results
Twenty-six metabolites were identified and quantified in the EMI, nine metabolites reflected the sex of the embryo rather than their presence.
Conclusions
1H NMR is sensitive enough to perform quantitative analysis of sex-induced differences in the EMI. These results may help to understand the embryo-maternal dialogue on the basis of embryonic sex.
References
Alonso, A., Marsal, S., & Julià, A. (2015). Analytical methods in untargeted metabolomics: state of the art in 2015. Frontiers in Bioengineering and Biotechnology,5(3), 23.
Bermejo-Alvarez, P., Rizos, D., Rath, D., Lonergan, P., & Gutierrez-Adan, A. (2008). Epigenetic differences between male and female bovine blastocysts produced in vitro. Physiological Genomics,32(2), 264–272.
Bracewell-Milnes, T., Saso, S., Abdalla, H., Nikolau, D., Norman-Taylor, J., Johnson, M., et al. (2017). Metabolomics as a tool to identify biomarkers to predict and improve outcomes in reproductive medicine: A systematic review. Human Reproduction Update.,23(6), 723–736.
Gómez, E., Caamaño, J. N., Corrales, F. J., Díez, C., Correia-Álvarez, E., Martín, D., et al. (2013). Embryonic sex induces differential expression of proteins in bovine uterine fluid. Journal of Proteome Research,12(3), 1199–1210.
Gómez, E., Carrocera, S., Martin, D., Herrero, P., Canela, N., & Muñoz, M. (2018a). Differential release of cell-signaling metabolites by male and female bovine embryos cultured in vitro. Theriogenology,114, 180–184.
Gómez, E., Martin, D., Carrocera, S., Sánchez-Calabuig, M. J., Gutierrez-Adán, A., Alonso-Guervos, M., et al. (2017). Expression and localization of ARTEMIN in the bovine uterus and embryos. Theriogenology,90, 153–162.
Gómez E, Muñoz M, Simó C, Ibáñez C, Carrocera S, Martín-González D, Cifuentes A (2016). Non-invasive metabolomics for improved determination of embryonic sex markers in chemically defined culture medium. Journal of Chromatography A. 1474, 138–144. Erratum in: Journal of Chromatography A. 2017.1526:167.
Gómez, E., Sánchez-Calabuig, M. J., Martin, D., Carrocera, S., Murillo, A., Correia-Alvarez, E., et al. (2018b). In vitro cultured bovine endometrial cells recognize embryonic sex. Theriogenology,108, 176–184.
Gross, N., Kropp, J., & Khatib, H. (2017). Sexual Dimorphism of miRNAs Secreted by Bovine In vitro-produced Embryos. Frontiers in Genetics,8, 39.
Laguna-Barraza, R., Bermejo-Álvarez, P., Ramos-Ibeas, P., de Frutos, C., López-Cardona, A. P., Calle, A., et al. (2013). Sex-specific embryonic origin of postnatal phenotypic variability. Reproduction, Fertility, and Development,25(1), 38–47.
Lamy, J., Gatien, J., Dubuisson, F., Nadal-Desbarats, L., Salvetti, P., Mermillod, P., et al. (2018). Metabolomic profiling of bovine oviductal fluid across the oestrous cycle using proton nuclear magnetic resonance spectroscopy. Reproduction, Fertility, and Development,30(7), 1021–1028.
Rubessa, M., Ambrosi, A., Gonzalez-Pena, D., Polkoff, K. M., & Wheeler, M. B. (2018). Non-invasive nuclear magnetic resonance analysis of male and female embryo metabolites during in vitro embryo culture. Metabolomics,14(9), 113.
Sponchiado, M., Gonella-Diaza, A. M., Rocha, C. C., Turco, E. G. L., Pugliesi, G., Leroy, J. L. M. R., et al. (2019). The pre-hatching bovine embryo transforms the uterine luminal metabolite composition in vivo. Scientific Reports,9(1), 8354.
Sturmey, R. G., Bermejo-Alvarez, P., Gutierrez-Adan, A., Rizos, D., Leese, H. J., & Lonergan, P. (2010). Amino acid metabolism of bovine blastocysts: A biomarker of sex and viability. Molecular Reproduction and Development,77(3), 285–296.
Trigal, B., Gomez, E., Díez, C., Caamaño, J. N., Muñoz, M., Moreno, J. F., et al. (2012). Comparative study of PCR-sexing procedures using bovine embryos fertilized with sex-sorted spermatozoa. Spanish Journal of Agricultural Research,10, 353–359.
Acknowledgements
The authors wish to thank JN Caamaño for his assistance in cell culture and Ann Hannigan for the English revision of this article. Spanish Ministry of Economy and Competitiveness (MINECO) projects AGL2016-78597R and AGL2016-81890-REDT (Redes de Excelencia). Fondo Europeo de Desarrollo Regional (FEDER) and M. Velázquez (Newcastle University, UK), whose critical discussion is very appreciated. The authors are members of the COST Action 16119, In vitro 3-D Total Cell guidance and Fitness (Cellfit).
Author information
Authors and Affiliations
Contributions
EG and MM conceived and designed research. DMG, SC, JG and PS conducted experiments. PS and JG contributed new reagents or analytical tools. EG analysed data. MM wrote the manuscript. All authors read and approved the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
None of the authors have any conflicts of interest.
Ethical approval
All experimental procedures were approved by the Animal Research Ethics Committee of SERIDA (Agreement 02/02/2012), in accordance with the EC Directive 86/609/EC.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Muñoz, M., Gatien, J., Salvetti, P. et al. Nuclear magnetic resonance analysis of female and male pre-hatching embryo metabolites at the embryo-maternal interface. Metabolomics 16, 47 (2020). https://doi.org/10.1007/s11306-020-01672-4
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11306-020-01672-4