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BMC veterinary research2024; 20(1); 272; doi: 10.1186/s12917-024-04129-1

Preovulatory follicular fluid secretome added to in vitro maturation medium influences the metabolism of equine cumulus-oocyte complexes.

Abstract: In vitro embryo production is a highly demanded reproductive technology in horses, which requires the recovery (in vivo or post-mortem) and in vitro maturation (IVM) of oocytes. Oocytes subjected to IVM exhibit poor developmental competence compared to their in vivo counterparts, being this related to a suboptimal composition of commercial maturation media. The objective of this work was to study the effect of different concentrations of secretome obtained from equine preovulatory follicular fluid (FF) on cumulus-oocyte complexes (COCs) during IVM. COCs retrieved in vivo by ovum pick up (OPU) or post-mortem from a slaughterhouse (SLA) were subjected to IVM in the presence or absence of secretome (Control: 0 µg/ml, S20: 20 µg/ml or S40: 40 µg/ml). After IVM, the metabolome of the medium used for oocyte maturation prior (Pre-IVM) and after IVM (Post-IVM), COCs mRNA expression, and oocyte meiotic competence were analysed. Results: IVM leads to lactic acid production and an acetic acid consumption in COCs obtained from OPU and SLA. However, glucose consumption after IVM was higher in COCs from OPU when S40 was added (Control Pre-IVM vs. S40 Post-IVM: 117.24 ± 7.72 vs. 82.69 ± 4.24; Mean µM ± SEM; p < 0.05), while this was not observed in COCs from SLA. Likewise, secretome enhanced uptake of threonine (Control Pre-IVM vs. S20 Post-IVM vs. S40 Post-IVM: 4.93 ± 0.33 vs. 3.04 ± 0.25 vs. 2.84 ± 0.27; Mean µM ± SEM; p < 0.05) in COCs recovered by OPU. Regarding the relative mRNA expression of candidate genes related to metabolism, Lactate dehydrogenase A (LDHA) expression was significantly downregulated when secretome was added during IVM at 20-40 µg/ml in OPU-derived COCs (Control vs. S20 vs. S40: 1.77 ± 0.14 vs. 1 ± 0.25 vs. 1.23 ± 0.14; fold change ± SEM; p < 0.05), but not in SLA COCs. Conclusions: The addition of secretome during in vitro maturation (IVM) affects the gene expression of LDHA, glucose metabolism, and amino acid turnover in equine cumulus-oocyte complexes (COCs), with diverging outcomes observed between COCs retrieved using ovum pick up (OPU) and slaughterhouse-derived COCs (SLA).
© 2024. The Author(s).
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Publication Date: 2024-06-25 PubMed ID: 38918770PubMed Central: PMC11197253DOI: 10.1186/s12917-024-04129-1Google Scholar: Lookup
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Summary

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This research investigates the effects of adding preovulatory follicular fluid secretome to the maturation medium of horse oocytes during in vitro maturation. The study found a difference in gene expression, glucose metabolism, and amino acid turnover between oocytes collected via ovum pick up (OPU) and those sourced from a slaughterhouse.

Study Background

  • The research focuses on enhancing in-vitro embryo production in horses; a process that involves recovering and maturing oocytes in vitro (IVM).
  • Commercially available maturation media have been reported as suboptimal for this process, leading to poor developmental competence in oocytes subjected to IVM compared to those matured in vivo.
  • The researchers sought to improve this by supplementing the IVM medium with secretome obtained from equine preovulatory follicular fluid (FF).

Study Design

  • The secretome was added to the maturation medium at different concentrations (0, 20, and 40 µg/ml).
  • Cumulus-oocyte complexes (COCs) used for the study were either collected via ovum pick up (OPU) or sourced post-mortem from a slaughterhouse (SLA).
  • The effects of this treatment on the metabolome of the used maturation medium, the COCs’ mRNA expression, and oocyte meiotic competence were assessed.

Study Findings

  • The research found that IVM resulted in lactic acid production and acetic acid consumption in the COCs, regardless of their source.
  • However, glucose consumption after IVM was higher in OPU-sourced COCs when higher concentrations of secretome were added.
  • Higher secretome concentrations also led to increased uptake of the amino acid threonine in OPU-sourced COCs.
  • Gene expression of Lactate dehydrogenase A (LDHA), which is related to metabolism, was prominently downregulated in the presence of secretome among the OPU-derived COCs.
  • No significant changes in glucose metabolism, threonine uptake, or gene expression were observed in SLA-sourced COCs with or without added secretome.

Conclusion

  • The addition of preovulatory follicular fluid secretome to the IVM medium appears to affect gene expression, glucose metabolism, and amino acid turnover in COCs, but the effects differ based on the source of COCs.
  • Further research may be required to understand why the source of the COC has such a significant impact on the effectiveness of the supplementation.

Cite This Article

APA
Luis-Calero M, Ortiz-Rodríguez JM, Fernández-Hernández P, Muñoz-García CC, Pericuesta E, Gutiérrez-Adán A, Marinaro F, Embade N, Conde R, Bizkarguenaga M, Millet Ó, González-Fernández L, Macías-García B. (2024). Preovulatory follicular fluid secretome added to in vitro maturation medium influences the metabolism of equine cumulus-oocyte complexes. BMC Vet Res, 20(1), 272. https://doi.org/10.1186/s12917-024-04129-1

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 20
Issue: 1
Pages: 272
PII: 272

Researcher Affiliations

Luis-Calero, Marcos
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Facultad de Veterinaria, Instituto de Investigación INBIO G+C, Universidad de Extremadura, Av. de la Universidad s/n, Cáceres, 10004, Spain.
Ortiz-Rodríguez, José Manuel
  • Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Via Tolara di Sopra 50, Bologna, 40064, Ozzano dell'Emilia, Italy.
Fernández-Hernández, Pablo
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Facultad de Veterinaria, Instituto de Investigación INBIO G+C, Universidad de Extremadura, Av. de la Universidad s/n, Cáceres, 10004, Spain.
Muñoz-García, Carmen Cristina
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Facultad de Veterinaria, Instituto de Investigación INBIO G+C, Universidad de Extremadura, Av. de la Universidad s/n, Cáceres, 10004, Spain.
Pericuesta, Eva
  • Departamento de Reproducción Animal, INIA-CSIC, Av. Puerta de Hierro 18, Madrid, 28040, Spain.
Gutiérrez-Adán, Alfonso
  • Departamento de Reproducción Animal, INIA-CSIC, Av. Puerta de Hierro 18, Madrid, 28040, Spain.
Marinaro, Federica
  • Departamento de Reproducción Animal, INIA-CSIC, Av. Puerta de Hierro 18, Madrid, 28040, Spain.
Embade, Nieves
  • Precision Medicine and Metabolism Laboratory, CIC bioGUNE, Basque Research and Technology Alliance (BRTA), Parque Tecnológico de Bizkaia, 801 A Building, Derio, 48160, Bizkaia, Spain.
Conde, Ricardo
  • Precision Medicine and Metabolism Laboratory, CIC bioGUNE, Basque Research and Technology Alliance (BRTA), Parque Tecnológico de Bizkaia, 801 A Building, Derio, 48160, Bizkaia, Spain.
Bizkarguenaga, Maider
  • Precision Medicine and Metabolism Laboratory, CIC bioGUNE, Basque Research and Technology Alliance (BRTA), Parque Tecnológico de Bizkaia, 801 A Building, Derio, 48160, Bizkaia, Spain.
Millet, Óscar
  • Precision Medicine and Metabolism Laboratory, CIC bioGUNE, Basque Research and Technology Alliance (BRTA), Parque Tecnológico de Bizkaia, 801 A Building, Derio, 48160, Bizkaia, Spain.
González-Fernández, Lauro
  • Departamento de Bioquímica y Biología Molecular y Genética, Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Av. de la Universidad s/n, Cáceres, 10004, Spain.
Macías-García, Beatriz
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Facultad de Veterinaria, Instituto de Investigación INBIO G+C, Universidad de Extremadura, Av. de la Universidad s/n, Cáceres, 10004, Spain. bemaciasg@unex.es.

MeSH Terms

  • Animals
  • Horses
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Follicular Fluid / metabolism
  • Follicular Fluid / chemistry
  • In Vitro Oocyte Maturation Techniques / veterinary
  • Cumulus Cells / metabolism
  • Cumulus Cells / drug effects
  • Female
  • Culture Media / pharmacology
  • Secretome / metabolism

Grant Funding

  • RC1 / Banco Santander
  • J45F21002000001 / Ministero dell'Universitu00e0 e della Ricerca
  • IB20005 / Junta de Extremadura
  • PI-0152-22 / Junta de Extremadura
  • IB20005 / Junta de Extremadura
  • PID2021-122507OB-I00 / Ministerio de Ciencia e Innovaciu00f3n
  • FJC2021-047675-I / Ministerio de Ciencia e Innovaciu00f3n
  • PID2020-112723RBI00 / Ministerio de Ciencia e Innovaciu00f3n
  • PID2020-112723RBI00 / Ministerio de Ciencia e Innovaciu00f3n

Conflict of Interest Statement

The authors declare no competing interests.

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