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Theriogenology2020; 160; 40-49; doi: 10.1016/j.theriogenology.2020.10.036

Mitochondrial function, blastocyst development and live foals born after ICSI of immature vitrified/warmed equine oocytes matured with or without melatonin.

Abstract: Oocyte vitrification is considered experimental in the horse with only three live foals reported. The oxidative conditions induced by vitrification could in part explain the poor results and melatonin, a powerful antioxidant, could stimulate ROS metabolization and restore mitochondrial function in these oocytes. Our objective was to determine the oxidative status of vitrified equine oocytes and to analyze the effect of melatonin on mitochondrial-specific ROS (mROS), oocyte maturation, ICSI embryo development and viability. Immature, abattoir-derived oocytes were held for 15 h and vitrified in a final concentration of 20% EG, 20% DMSO and 0.65 M trehalose. In Experiment 1, overall ROS was determined by DCHF-DA; vitrification increased ROS production compared to non-vitrified controls (1.29 ± 0.22 vs 0.74 ± 0.25 a. u.; P = 0.0156). In Experiment 2, mROS was analyzed by MitoSOX™ in vitrified/warmed oocytes matured with (+) or without (-) supplementation of 10-9 M melatonin; mROS decreased in vitrified and non-vitrified oocytes matured in presence of melatonin (P < 0.05). In Experiment 3, we assessed the effect of melatonin supplementation on oocyte maturation, embryo development after ICSI, and viability by pregnancy establishment. Melatonin did not improve oocyte maturation, cleavage or blastocyst rate of non-vitrified oocytes. However, vitrified melatonin (+) oocytes reached similar cleavage (61, 75 and 77%, respectively) and blastocyst rate (15, 29 and 26%, respectively) than non-vitrified, melatonin (+) and (-) oocytes. Vitrified, melatonin (-) oocytes had lower cleavage (46%) and blastocyst rate (9%) compared to non-vitrified groups (P < 0.05), but no significant differences were observed when compared to vitrified melatonin (+). Although the lack of available recipients precluded the transfer of every blastocyst produced in our study, transferred embryos from non-vitrified oocytes resulted in 50 and 83% pregnancy rates while embryos from vitrified oocytes resulted in 17 and 33% pregnancy rates, from melatonin (+) and (-) treatments respectively. Two healthy foals, one colt from melatonin (+) and one filly from melatonin (-) treatment, were born from vitrified/warmed oocytes. Gestation lengths (considering day 0 = day of ICSI) were 338 days for the colt and 329 days for the filly, respectively. Our work showed for the first time that in the horse, as in other species, intracellular reactive oxygen species are increased by the process of vitrification. Melatonin was useful in reducing mitochondrial-related ROS and improving ICSI embryo development, although the lower pregnancy rate in presence of melatonin should be further analyzed in future studies. To our knowledge this is the first report of melatonin supplementation to an in vitro embryo culture system and its use to improve embryo developmental competence of vitrified oocytes following ICSI.
Copyright © 2020 Elsevier Inc. All rights reserved.
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Publication Date: 2020-11-02 PubMed ID: 33171351DOI: 10.1016/j.theriogenology.2020.10.036Google 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 research explored whether the antioxidant melatonin could boost the success rate of horse egg (oocyte) vitrification, a preservation technique. The research revealed that melatonin indeed reduced oxidative stress in vitrified oocytes and improved embryo development, leading to the birth of healthy foals. However, the impact of melatonin on pregnancy rates warrants further studies.

Objective and Methodology

  • The research aimed to understand how vitrification impacts the oxidative status of horse oocytes, and whether melatonin could help restore their mitochondrial function and improve their maturation and viability.
  • The eggs used for the study were obtained from a slaughterhouse. These immature eggs were preserved using vitrification and then exposed to melatonin.
  • The effects of vitrification and melatonin were assessed through several experiments that evaluated parameters such as Reactive Oxygen Species (ROS) production, mitochondrial-specific ROS (mROS), oocyte maturation, fertilized egg (embryo) development, and pregnancy establishment.

Findings

  • The findings confirmed that vitrification induced oxidative stress in the eggs, shown by an increase in overall ROS production as compared to non-vitrified eggs.
  • Supplementation of melatonin reduced mROS in both vitrified and non-vitrified eggs. This suggests that melatonin could counteract the oxidative stress induced by vitrification.
  • While melatonin did not improve the maturation or development rates of non-vitrified eggs, it significantly improved the outcomes for vitrified eggs. Vitrified eggs exposed to melatonin were almost as successful as non-vitrified ones in terms of their cleavage and development into blastocysts.
  • Importantly, two healthy foals were born from vitrified eggs subjected to melatonin treatment. This represents a direct, positive outcome of the addition of this antioxidant.

Implications

  • This research demonstrates, for the first time, that melatonin could be a vital supplement in the vitrification process of horse oocytes. It shows the potential of melatonin in mitigating oxidative damage and enhancing embryo development, increasing success rates of in vitro fertilization using vitrified eggs.
  • Despite these promising results, the study also revealed a lower pregnancy rate in the presence of melatonin. Therefore, more research is required to understand the impact and optimize the use of melatonin in in vitro embryo culture systems.

Cite This Article

APA
Clérico G, Taminelli G, Veronesi JC, Polola J, Pagura N, Pinto C, Sansinena M. (2020). Mitochondrial function, blastocyst development and live foals born after ICSI of immature vitrified/warmed equine oocytes matured with or without melatonin. Theriogenology, 160, 40-49. https://doi.org/10.1016/j.theriogenology.2020.10.036

Publication

Theriogenology
ISSN: 1879-3231
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 160
Pages: 40-49

Researcher Affiliations

Clérico, G
  • Facultad de Ingeniería y Ciencias Agrarians, Universidad Católica Argentina, 1107, Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.
Taminelli, G
  • Facultad de Ingeniería y Ciencias Agrarians, Universidad Católica Argentina, 1107, Buenos Aires, Argentina.
Veronesi, J C
  • Frigorífico Lamar, 6600, Buenos Aires, Argentina.
Polola, J
  • Haras La Aguada El Dok, 6708, Buenos Aires, Argentina; Facultad de Ciencias Veterinarias, Universidad Nacional de Rosario, 2170, Santa Fe, Argentina.
Pagura, N
  • Facultad de Ciencias Veterinarias, Universidad Nacional de Rosario, 2170, Santa Fe, Argentina.
Pinto, C
  • Louisiana State School of Veterinary Medicine, 70808, Baton Rouge, LA, USA.
Sansinena, M
  • Facultad de Ingeniería y Ciencias Agrarians, Universidad Católica Argentina, 1107, Buenos Aires, Argentina; Louisiana State School of Veterinary Medicine, 70808, Baton Rouge, LA, USA; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina. Electronic address: msansinena@uca.edu.ar.

MeSH Terms

  • Animals
  • Blastocyst
  • Cryopreservation / veterinary
  • Embryonic Development
  • Female
  • Horses
  • Male
  • Melatonin / pharmacology
  • Mitochondria
  • Oocytes
  • Pregnancy
  • Sperm Injections, Intracytoplasmic / veterinary
  • Vitrification

Citations

This article has been cited 20 times.
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