Time-lapse imaging and developmental competence of donkey eggs after ICSI: Effect of preovulatory follicular fluid during oocyte in vitro maturation.
Abstract: Equus members exhibit very divergent karyotype, genetic plasticity, and significant differences in their reproductive physiology. Despite the fact that somatic cell nuclear transfer and intracytoplasmic sperm injection (ICSI) has gained relevance in the last few years in horses, few reports have been published exploring ovum pick up (OPU) and in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) in donkeys. Yet, some donkey species and breeds are considered endangered, and these assisted-reproductive technologies could help to preserve the genetic of valuable individuals. In this study, we tested the hypothesis that supplementation with jenny preovulatory follicular fluid (PFF) during IVM could improve oocyte developmental competence in the donkey. For this, in vitro nuclear maturation rates, cumulus cell expansion, and embryo development after ICSI of donkey COCs matured in culture media supplemented with fetal bovine serum (FBS) or donkey PFF, with a known metabolomic profile, were assessed. Time-lapse imagining was performed after ICSI of horse and donkey oocytes. Eight OPU sessions were done in five jennies with an average recovery rate of 69.2% (n = 45 COCs). Although lower cumulus cells expansion was observed in oocytes of PFF group (P = 0.0010), no significant differences were described in nuclear maturation rates and preimplantation embryo development between groups. Donkey ICSI embryos showed similar morphokinetics to horse ICSI embryos. Our study shows that supplementing IVM media with FBS or donkey PFF supports nuclear maturation and early preimplantation embryo development after ICSI in donkeys. To our knowledge, the present study is the first report of ICSI, time-lapse imaging and in vitro blastocyst production in donkey.
Copyright © 2022 Elsevier Inc. All rights reserved.
Publication Date: 2022-11-01 PubMed ID: 36356490DOI: 10.1016/j.theriogenology.2022.10.030Google Scholar: Lookup
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- Journal Article
Summary
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This research explores the effect of using preovulatory follicular fluid (PFF) during the in vitro maturation of donkey (jenny) eggs, specifically in relation to their maturation rate, cell expansion and embryo development after Intracytoplasmic Sperm Injection (ICSI). The study found that supplementing the maturation process with donkey PFF or fetal bovine serum (FBS) supports nuclear maturation and early preimplantation embryo development.
Objective of the Research
- The objective of this study was to examine the impact of supplementing in vitro maturation with preovulatory follicular fluid (PFF) obtained from female donkeys (known as jennies) on the developmental competence of donkey eggs.
- The researchers aimed to test whether the practice could increase the success rate of the in vitro maturation process, with a focus on nuclear maturation rates, cumulus cell expansion, and post-ICSI embryo development.
- Underpinning the research is the increasing importance of assisted reproductive technologies in maintaining the genetic diversity of donkey species and breeds, some of which are considered endangered.
Methodology
- The researchers compared the effects of using fetal bovine serum (FBS) and donkey PFF during in vitro maturation.
- Time-lapse imaging was conducted following the ICSI of donkey and horse oocytes.
- The team conducted eight ovum pick-up sessions from five jennies, yielding a total of 45 cumulus-oocyte complexes (COCs).
Findings
- Contrary to their hypothesis, the researchers found that donkey COCs matured in culture media supplemented with either FBS or donkey PFF showed no significant differences in nuclear maturation rates and early preimplantation embryo development post-ICSI.
- While the cumulus cells of the PFF group showed lower expansion, this did not seem to affect overall maturation and development.
- ICSI embryos from donkeys exhibited similar morphokinetics to those extracted from horses after ICSI.
Significance
- This study is the first to report on ICSI, time-lapse imaging and in vitro blastocyst production in the donkey.
- The findings suggest that both FBS and donkey PFF can effectively supplement the in vitro maturation process, offering potential routes to preserving the genetic diversity of threatened donkey species and breeds.
Cite This Article
APA
Flores Bragulat AP, Ortiz I, Catalán J, Dorado J, Hidalgo M, Losinno L, Dordas-Perpinyà M, Yánez-Ortiz I, Martínez-Rodero I, Miró J, Gambini A.
(2022).
Time-lapse imaging and developmental competence of donkey eggs after ICSI: Effect of preovulatory follicular fluid during oocyte in vitro maturation.
Theriogenology, 195, 199-208.
https://doi.org/10.1016/j.theriogenology.2022.10.030 Publication
Researcher Affiliations
- Laboratorio de Producción Equina, Facultad de Agronomía y Veterinaria, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina.
- Veterinary Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, 14014, Cordoba, Spain.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, 08193, Bellaterra, Barcelona, Spain.
- Veterinary Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, 14014, Cordoba, Spain.
- Veterinary Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, 14014, Cordoba, Spain.
- Laboratorio de Producción Equina, Facultad de Agronomía y Veterinaria, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, 08193, Bellaterra, Barcelona, Spain.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, 08193, Bellaterra, Barcelona, Spain.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, 08193, Bellaterra, Barcelona, Spain.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, 08193, Bellaterra, Barcelona, Spain. Electronic address: jordi.miro@uab.cat.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina; Veterinary Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, 14014, Cordoba, Spain; School of Agriculture and Food Sciences, The University of Queensland, 4343, Gatton, Queensland, Australia. Electronic address: a.gambini@uq.edu.au.
MeSH Terms
- Male
- Pregnancy
- Animals
- Female
- Horses
- In Vitro Oocyte Maturation Techniques / veterinary
- Follicular Fluid
- Equidae
- Time-Lapse Imaging / veterinary
- Sperm Injections, Intracytoplasmic / veterinary
- Semen
Conflict of Interest Statement
Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Citations
This article has been cited 2 times.- Gambini A, Smith JM, Gurkin RJ, Palacios PD. Current and Emerging Advanced Techniques for Breeding Donkeys and Mules. Animals (Basel) 2025 Mar 29;15(7).
- Catalán J, Padilla L, Maside C, Martínez-Hernández J, Yánez-Ortiz I, Tvarijonaviciute A, Barranco I, Bonet S, Miró J, Yeste M. Redox profiling of preovulatory follicular fluid in the donkey is species-specific, and contributes to modulate sperm function. Sci Rep 2025 Feb 23;15(1):6522.
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