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Equine veterinary journal2020; 53(4); 787-795; doi: 10.1111/evj.13342

A high glucose concentration during early stages of in vitro equine embryo development alters expression of genes involved in glucose metabolism.

Abstract: Equine embryos exhibit an unusual pattern of glucose tolerance in vitro and are currently cultured in hyperglycaemic conditions. Objective: Our main objective was to analyse the effect of different glucose concentrations on in vitro-produced equine embryo development and quality. Methods: Experiments comparing in vitro and in vivo produced embryos. Methods: Oocytes (n = 641) were collected from post-mortem ovaries, matured in vitro and fertilised by intracytoplasmic sperm injection (ICSI). Embryo culture was divided from Day 0 to Day 4 and from Day 4 to Day 9 in three groups: 5-10 (5 and 10 mmol/L glucose respectively; n = 87); 5-17 (5 and 17.5 mmol/L; n = 66); and 10-17 (10 and 17.5 mmol/L; n = 117). A control group of 20 in vivo produced blastocysts was included. Cleavage and blastocyst rates were evaluated and embryos were snap-frozen for analysis of the relative mRNA expression of genes related to mitochondrial function, DNA methylation, apoptosis, glucose transport and metabolism. Results: No differences were observed in the cleavage or blastocyst rates among in vitro groups. Under high glucose conditions in vitro (10-17 group), BAX/BCL2 was higher, and PFKP, LDHA and COX2 were overexpressed compared to all other groups. The two groups with 5 mmol/L glucose concentration during the first culture stage (5-10 and 5-17) displayed similar patterns which differed to the 10-17 group. Conclusions: Conclusions related to embryo quality are based on gene expression patterns. Transfer of in vitro-produced embryos would reveal whether the observed differences improve embryo developmental competence. Conclusions: Five mM glucose during the first days of culture seems to be preferable to avoid over-activation of embryonic glycolytic pathways. Further studies are necessary to determine whether this improves embryo developmental competence.
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Publication Date: 2020-10-12 PubMed ID: 32881040DOI: 10.1111/evj.13342Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The researchers’ study suggests that high glucose concentrations during the early stages of equine embryo development in a lab environment can change the expression of genes related to glucose metabolism. However, further studies are needed to see if these changes can improve the development of embryos.

Study Objective

The main goal of this study was to examine how different glucose concentrations impact the development and quality of equine embryos cultivated in vitro.

Methods

  • To carry out this analysis, 641 oocytes (immature egg cells) were obtained from equine ovaries after death.
  • These were then matured in a lab setting and fertilized through the process of intracytoplasmic sperm injection (ICSI).
  • The researchers divided the embryos’ cultivation period into two stages – from Day 0 to Day 4 and from Day 4 to Day 9. These embryos were further divided into three sub-groups: 5-10 (5 and 10 mmol/L glucose respectively; n = 87), 5-17 (5 and 17.5 mmol/L; n = 66), and 10-17 (10 and 17.5 mmol/L; n = 117).
  • A control group of 20 in vivo produced blastocysts was included to compare to the in vitro groups.
  • Fertilization and blastocyst rates were assessed and embryos were frozen to analyze the relative mRNA expression of genes associated with mitochondrial function, DNA methylation, apoptosis (cell death), glucose transport and metabolism.

Results

  • They didn’t find any significant differences in fertilization or blastocyst rates among the in vitro groups.
  • In the 10-17 group, which was exposed to high glucose conditions in vitro, certain genes (BAX/BCL2, PFKP, LDHA, and COX2) were overexpressed compared to all other groups.
  • The two groups with 5 mmol/L glucose concentration during the first culture stage, (5-10 and 5-17), demonstrated similar gene expression patterns, which were different from the 10-17 group.

Conclusions

  • The conclusions about embryo quality were based on the patterns of gene expression observed during the study.
  • The differences in gene expression suggest that in vitro-produced embryos may see improved development, but this has yet to be confirmed via the successful transfer of these embryos.
  • It appears that a 5 mM glucose concentration during the early days of culture seems to be optimal to avoid over-stimulating embryonic glycolytic pathways.
  • The authors emphasize the need for further studies to confirm whether this specific glucose concentration during early cultivation stages can enhance embryo developmental competence.

Cite This Article

APA
Sánchez-Calabuig MJ, Fernández-González R, Hamdi M, Smits K, López-Cardona AP, Serres C, Macías-García B, Gutiérrez-Adán A. (2020). A high glucose concentration during early stages of in vitro equine embryo development alters expression of genes involved in glucose metabolism. Equine Vet J, 53(4), 787-795. https://doi.org/10.1111/evj.13342

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 53
Issue: 4
Pages: 787-795

Researcher Affiliations

Sánchez-Calabuig, María J
  • Department of Animal Medicine and Surgery, Faculty of Veterinary Science, University Complutense of Madrid, Madrid, Spain.
  • Department of Animal Reproduction, INIA, Madrid, Spain.
Fernández-González, Raúl
  • Department of Animal Reproduction, INIA, Madrid, Spain.
Hamdi, Meriem
  • Department of Animal Reproduction, INIA, Madrid, Spain.
Smits, Katrien
  • Department of Obstetrics, Reproduction and Herd Health, Ghent University, Ghent, Belgium.
López-Cardona, Angela P
  • Department of Animal Reproduction, INIA, Madrid, Spain.
  • Grupo de Investigación (GI) - Biogénesis, Universidad de Antioquia, Medellín, Colombia.
Serres, Consuelo
  • Department of Animal Medicine and Surgery, Faculty of Veterinary Science, University Complutense of Madrid, Madrid, Spain.
Macías-García, Beatriz
  • Animal Medicine Department, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain.
  • Research Group of Intracellular Signalling and Technology of Reproduction (SINTREP), Research Institute of Biotechnology in Livestock and Cynegetic (INBIO G+C), University of Extremadura, Cáceres, Spain.
Gutiérrez-Adán, Alfonso
  • Department of Animal Reproduction, INIA, Madrid, Spain.

MeSH Terms

  • Animals
  • Blastocyst
  • Embryo Culture Techniques / veterinary
  • Embryonic Development
  • Glucose
  • Horses
  • Oocytes
  • Sperm Injections, Intracytoplasmic / veterinary

Grant Funding

  • AGL2017-84681-R / Spanish Ministry of Science, Innovation and Universities Competitiveness
  • RTI2018-093548 / Ministerio de Ciencia e Innovación
  • RYC-2017-21545 / Fondo Europeo de Desarrollo Regional (FEDER)
  • 512/2010 / Universidad de Antioquia (COLCIENCIAS)

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Citations

This article has been cited 5 times.
  1. Wu D, Liu C, Ding L. Follicular metabolic dysfunction, oocyte aneuploidy and ovarian aging: a review. J Ovarian Res 2025 Mar 12;18(1):53.
    doi: 10.1186/s13048-025-01633-2pubmed: 40075456google scholar: lookup
  2. Zhao H, Sun Y, Feng H, Cai J, Liu Y, Li Y, Chen S, Zhou Z, Du Y, Zeng X, Ren H, Su W, Mei Q, Chen G. PFKP silencing suppresses tumor growth via the AXL-MET axis. Int J Biol Sci 2024;20(15):6056-6072.
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  3. Wang Y, Guo Y, Duan C, Yang R, Zhang L, Liu Y, Zhang Y. Long Non-Coding RNA GDAR Regulates Ovine Granulosa Cells Apoptosis by Affecting the Expression of Apoptosis-Related Genes. Int J Mol Sci 2022 May 6;23(9).
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  4. Uysal F, Kahveci S, Sukur G, Cinar O. Embryo culture media differentially alter DNA methylating enzymes and global DNA methylation in embryos and oocytes. J Mol Histol 2022 Feb;53(1):63-74.
    doi: 10.1007/s10735-021-10038-6pubmed: 34741214google scholar: lookup
  5. Cañón-Beltrán K, Cajas YN, Peréz-Cerezales S, Leal CLV, Agirregoitia E, Gutierrez-Adán A, González EM, Rizos D. Nobiletin enhances the development and quality of bovine embryos in vitro during two key periods of embryonic genome activation. Sci Rep 2021 Jun 3;11(1):11796.
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