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Non-invasive nuclear magnetic resonance analysis of male and female embryo metabolites during in vitro embryo culture

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Abstract

Introduction

In the past 20+ years, several studies of bovine embryo production showed how the ratio of male to female embryos changes if embryos are made in vivo or in vitro. It is known that in in vitro systems, the sex ratio is in favor of males when there are high levels of glucose, and favors females when the principal energetic substrate is one other than glucose, like citrate.

Objectives

The aim of this study was to evaluate the embryo metabolism during three important periods of in vitro development: the early development (from day 1 until day 3), the middle of culture (day 3 until day 5), and later development (day 5 until day 7).

Methods

To obtain this information we evaluated the spent medium from each time period by 1H NMR.

Results

Our results confirm that embryo metabolism is different between sexes. The new information obtained by identifies markers that we can use to predict the embryo sex.

Conclusion

These results open a new, non-invasive method to evaluate sex of the embryos before the transfer. In the first period of embryo culture, valine concentration is good indicator (66.7% accurate), while in the last phase of culture, pyruvate depletion is the best marker (64% accurate) to evaluate the sex of the embryo.

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Acknowledgements

The authors would like to thank Professor Scott E. Denmark for this helpful discussions, the generous use of his lab and the 750-MHz Agilent VNS750NB spectrometer. This work was partially funded by the University of Illinois Experiment Station and the U.S. Department of Agriculture-National Institute of Food and Agriculture via Multistate Research Project W-2171, #ILLU-538-347.

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Authors and Affiliations

  1. Institute for Genomic Biology, University of Illinois, Urbana, IL, USA

    Marcello Rubessa & Matthew B. Wheeler

  2. Department of Animal Sciences, University of Illinois, 1207 West Gregory Drive, Urbana, IL, 61801, USA

    Marcello Rubessa, Dianelys Gonzalez-Pena, Kathryn M. Polkoff & Matthew B. Wheeler

  3. Department of Chemistry, University of Illinois, Urbana, IL, USA

    Andrea Ambrosi

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  1. Marcello Rubessa
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  2. Andrea Ambrosi
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Contributions

Conceived and designed the experiments: MR, MBW; Performed the experiments: MR, AA KMP, MBW; Analyzed the data: DG-P, MR, MBW; Contributed reagents/materials/analysis tools: DG-P, AA, Scott Denmark. Wrote the paper: MR, KMP, MBW. Critical revised the paper: MR, AA, DG-P, KMP, MBW.

Corresponding author

Correspondence to Matthew B. Wheeler.

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The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants/tissues or live animals performed by any of the authors. The oocytes were purchased from a commercial company, DeSoto Biosciences, Seymour, TN, USA. The oocytes were purchased by DeSoto Biosciences who aspirated the oocytes from the ovaries and provided them to us. The abattoir where the ovaries were purchased is a USDA inspected facility and conforms to all USDA regulations. We did not used any live animals for these studies. Similarly, the frozen semen used was from a commercial semen vendor. We purchased the semen used it these studies and had no contact with the bulls used for the in vitro fertilization. Our University does not require IACUC (animal care and use) protocols when using purchased tissues or abattoir materials. We therefore see no ethics issues with the present study.

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Rubessa, M., Ambrosi, A., Gonzalez-Pena, D. et al. Non-invasive nuclear magnetic resonance analysis of male and female embryo metabolites during in vitro embryo culture. Metabolomics 14, 113 (2018). https://doi.org/10.1007/s11306-018-1414-0

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