Noninvasive characterization of metabolites secreted in culture media by bovine embryos during in vitro production

Santos, E. C. dos; Lima, Camila Bruna de; Annes, Kelly; Milazzotto, Marcella Pécora

doi:10.1007/s11306-016-1029-2

Cited by 6 publications

(6 citation statements)

References 37 publications

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“…Previous reports stated numerous differences in the transcription pattern and DNA methylation profile between bovine embryos developing at different cleavage speeds 15,18 . Other studies provided evidences that such differences were related to the metabolic stress caused by the culture system 16,17 . Based on these assumptions, the present study was initially designed to investigate how Fast and Slow embryos respond to stressors commonly found during in vitro culture, i.e., non-optimized oxygen tension and high glucose concentration.…”

Section: Discussionmentioning

confidence: 98%

“…In the bovine, the kinetics of the first 2 cell divisions is associated with distinct phenotypes at the blastocyst stage 15 . Molecular and biochemical characterization demonstrated that fast-developing embryos exhibit improved ability to trigger adaptive responses 15 – 17 and distinct profiles of epigenetic reprogramming (more DNA methylation) 18 .…”

Section: Introductionmentioning

confidence: 99%

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The dynamics between in vitro culture and metabolism: embryonic adaptation to environmental changes

Lima

Santos

Ispada

et al. 2020

Sci Rep

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Previous studies have discussed the importance of an optimal range of metabolic activity during preimplantation development. To avoid factors than can trigger an undesirable trajectory, it is important to learn how nutrients and metabolites interact to help launching the correct developmental program of the embryo, and how much the in vitro culture system can impair this process. Here, using the bovine model, we describe a factorial experimental design used to investigate the biochemical and molecular signature of embryos in response to different combinations of morphological features—i.e. speed of development—and external stimuli during in vitro culture—i.e. different oxygen tensions and glucose supplementation. Our analyses demonstrate that the embryos present heterogeneous metabolic responses depending on early morphological phenotypes and the composition of their surroundings. However, despite the contribution of each single stimulus for the embryo phenotype, oxygen tension is determinant for such differences. The lower oxygen environment boosts the metabolism of embryos with faster kinetics, in particular those cultured in lower glucose concentrations.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

The dynamics between in vitro culture and metabolism: embryonic adaptation to environmental changes

Lima

Santos

Ispada

et al. 2020

Sci Rep

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“…Selection of embryos with higher potential to generate pregnancy is important to improve the success of IVP, and in this context, morphokinetics has been used as a promising tool. Recent studies showed that embryos with distinct kinetics of development present differences related to DNA damage [ 20 ], consumption of subtracts [ 10 ], secretion of metabolites [ 11 ], energy storage [ 10 ], activity of antioxidant enzymes, [ 21 ] cell survival [ 22 ], and transcription profile [ 10 ].…”

Section: Discussionmentioning

confidence: 99%

Genome-wide screening of DNA methylation in bovine blastocysts with different kinetics of development

Ispada

Lima

Sirard

et al. 2018

Epigenetics & Chromatin

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Background The timing of the first cell divisions may predict the developmental potential of an embryo, including its ability to establish pregnancy. Besides differences related to metabolism, stress, and survival, embryos with different speeds of development present distinct patterns of gene expression, mainly related to energy and lipid metabolism. As gene expression is regulated by epigenetic factors, and that includes DNA methylation patterns, in this study we compared the global DNA methylation profile of embryos with different kinetics of development in order to identify general pathways and regions that are most influenced by this phenotype. For this purpose, bovine embryos were in vitro produced using sexed semen (female), classified as fast (four or more cells) or slow (two cells) at 40 hpi and cultured until blastocyst stage, when they were analyzed.ResultsGenome-wide DNA methylation analysis identified 11,584 differently methylated regions (DMRs) (7976 hypermethylated regions in fast and 3608 hypermethylated regions in slow embryos). Fast embryos presented more regions classified as hypermethylated distributed throughout the genome, as in introns, exons, promoters, and repeat elements while in slow embryos, hypermethylated regions were more present in CpG islands. DMRs were clustered by means of biological processes, and the most affected pathways were related to cell survival/differentiation and energy/lipid metabolism. Transcripts profiles from DM genes connected with these pathways were also assessed, and the most part disclosed changes in relative quantitation.ConclusionThe kinetics of the first cleavages influences the DNA methylation and expression profiles of genes related to metabolism and differentiation pathways and may affect embryo viability.Electronic supplementary materialThe online version of this article (10.1186/s13072-017-0171-z) contains supplementary material, which is available to authorized users.

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“…Our group previously described that embryos completing the first two cleavage cycles earlier, that is, the faster ones, are remarkably different from those that need longer time for that (slower embryos). Such differences include the flow of metabolites and carbohydrates, mRNA profile, and genome-wide DNA methylation (dos Santos et al 2016, Silva et al 2016, Ispada et al 2018, 2020, de Lima et al 2020). The differences in kinetics are caused by follicular environment variations that lead to molecular changes in oocytes and, later, alteration in embryonic first cleavage dynamics, their metabolic status, and DNA methylation reprogramming (Alves et al 2019).…”

Section: Introductionmentioning

confidence: 99%

Metabolism-driven post-translational modifications of H3K9 in early bovine embryos

et al. 2021

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Metabolic and molecular profiles were reported as different for bovine embryos with distinct kinetics during the first cleavages. In this study, we used this same developmental model (fast vs slow) to determine if the relationship between metabolism and developmental kinetics affects the levels of acetylation or tri-methylation at histone H3 lysine 9 (H3K9ac and H3K9me3, respectively). Fast and slow developing embryos presented different levels of H3K9ac and H3K9me3 from the earliest stages of development (40 and 96 hpi) and up to the blastocyst stage. For H3K9me3, both groups of embryos presented a wave of demethylation and de novo methylation, although it was more pronounced in fast than slow embryos, resulting in blastocysts with higher levels of this mark. The H3K9ac reprogramming profile was distinct between kinetics groups. While slow embryos presented a wave of deacetylation, followed by an increase in this mark at the blastocyst stage, fast embryos reduced this mark throughout all the developmental stages studied. H3K9me3 differences corresponded to writer and eraser transcript levels, while H3K9ac patterns were explained by metabolism-related gene expression. To verify if metabolic differences could alter levels of H3K9ac, embryos were cultured with sodium-iodoacetate (IA) or dichloroacetate (DCA) to disrupt the glycolytic pathway or increase acetyl-CoA production, respectively. IA reduced H3K9ac while DCA increased H3K9ac in blastocysts. Concluding, H3K9me3 and H3K9ac patterns differ between embryos with different kinetics, the second one explained by metabolic pathways involved in acetyl-CoA production. So far, this is the first study demonstrating a relationship between metabolic differences and histone post-translational modifications in bovine embryos.

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Noninvasive characterization of metabolites secreted in culture media by bovine embryos during in vitro production

Cited by 6 publications

References 37 publications

The dynamics between in vitro culture and metabolism: embryonic adaptation to environmental changes

The dynamics between in vitro culture and metabolism: embryonic adaptation to environmental changes

Genome-wide screening of DNA methylation in bovine blastocysts with different kinetics of development

Metabolism-driven post-translational modifications of H3K9 in early bovine embryos

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