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Reproductive biology and endocrinology : RB&E2014; 12; 99; doi: 10.1186/1477-7827-12-99

Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential.

Abstract: Evaluation of mitochondrial function offers an alternative to evaluate embryo development for assessment of oocyte viability, but little information is available on the relationship between mitochondrial and chromatin status in equine oocytes. We evaluated these parameters in immature equine oocytes either fixed immediately (IMM) or held overnight in an Earle's/Hank's' M199-based medium in the absence of meiotic inhibitors (EH treatment), and in mature oocytes. We hypothesized that EH holding may affect mitochondrial function and that holding temperature may affect the efficiency of meiotic suppression. Methods: Experiment 1 - Equine oocytes processed immediately or held in EH at uncontrolled temperature (22 to 27°C) were evaluated for initial chromatin configuration, in vitro maturation (IVM) rates and mitochondrial energy/redox potential. Experiment 2 - We then investigated the effect of holding temperature (25°C, 30°C, 38°C) on initial chromatin status of held oocytes, and subsequently repeated mitochondrial energy/redox assessment of oocytes held at 25°C vs. immediately-evaluated controls. Results: EH holding at uncontrolled temperature was associated with advancement of germinal vesicle (GV) chromatin condensation and with meiotic resumption, as well as a lower maturation rate after IVM. Holding did not have a significant effect on mitochondrial distribution within chromatin configurations. Independent of treatment, oocytes having condensed chromatin had a significantly higher proportion of perinuclear/pericortical mitochondrial distribution than did other GV configurations. Holding did not detrimentally affect oocyte energy/redox parameters in viable GV-stage oocytes. There were no significant differences in chromatin configuration between oocytes held at 25°C and controls, whereas holding at higher temperature was associated with meiosis resumption and loss of oocytes having the condensed chromatin GV configuration. Holding at 25°C was not associated with progression of mitochondrial distribution pattern and there were no significant differences in oocyte energy/redox parameters between these oocytes and controls. Conclusions: Mitochondrial distribution in equine GV-stage oocytes is correlated with chromatin configuration within the GV. Progression of chromatin configuration and mitochondrial status during holding are dependent on temperature. EH holding at 25°C maintains meiotic arrest, viability and mitochondrial potential of equine oocytes. This is the first report on the effects of EH treatment on oocyte mitochondrial energy/redox potential.
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Publication Date: 2014-10-11 PubMed ID: 25306508PubMed Central: PMC4209075DOI: 10.1186/1477-7827-12-99Google 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 deals with the effect of holding equine oocytes (horse egg cells) in a specific medium before in vitro maturation, and the impact of different holding temperatures on meiotic suppression and mitochondrial function. The study suggests that these factors may affect whether or not these oocytes are viable.

Research Process

  • The scientists examined how holding immature equine oocytes in an Earle’s/Hank’s M199-based medium, devoid of meiotic inhibitors, impacts both chromatin and mitochondrial status, specifically the mitochondrial energy/redox potential.
  • The oocytes were either fixed immediately or held overnight with the experiment divided into two parts.
  • In the first experiment, the oocytes were held at an uncontrolled temperature (22 to 27°C) while in the second experiment, they were held at controlled temperatures of 25°C, 30°C, and 38°C.
  • After each experiment, the oocytes were evaluated for initial chromatin configuration, in vitro maturation (IVM) rates, and mitochondrial energy/redox potential.

Findings

  • The results showed that holding the oocytes at an uncontrolled temperature led to advancement of germinal vesicle (GV) chromatin condensation and meiotic resumption, and a reduced maturation rate post in vitro maturation (IVM).
  • The holding process did not significantly affect the distribution of mitochondria within various chromatin configurations.
  • Oocytes demonstrating the condensed form of chromatin had a higher proportion of mitochondria located near the nucleus (perinuclear) or towards the outer layer (pericortical) compared to other GV configurations.
  • The holding did not negatively impact the energy/redox parameters of viable GV-stage oocytes.

Temperature Specific Findings

  • When analyzing the effects of fixed holding temperatures, there were no significant differences in chromatin configuration between oocytes held at 25°C and the control group.
  • Holding the oocytes at higher temperatures resulted in the initiation of meiosis and a loss of oocytes presenting the condensed chromatin GV configuration.
  • Keeping the oocytes at 25°C did not lead to progression in the mitochondrial distribution pattern, and there were no significant differences in the oocyte energy/redox parameters between the 25°C samples and the control group.

Conclusions

  • The results of this research imply that in equine GV-stage oocytes, their chromatin configuration within the GV is correlated to the distribution of mitochondria.
  • Also, the progression of chromatin configuration and mitochondrial status while holding these oocytes is temperature dependent.
  • Lastly, it was found that holding at 25°C maintains meiotic arrest, as well as the viability and mitochondrial potential of equine oocytes, indicating that 25°C could be a suitable temperature for holding oocytes in the absence of meiotic inhibitors.

Cite This Article

APA
Martino NA, Dell'Aquila ME, Filioli Uranio M, Rutigliano L, Nicassio M, Lacalandra GM, Hinrichs K. (2014). Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential. Reprod Biol Endocrinol, 12, 99. https://doi.org/10.1186/1477-7827-12-99

Publication

Reproductive biology and endocrinology : RB&E
ISSN: 1477-7827
NlmUniqueID: 101153627
Country: England
Language: English
Volume: 12
Pages: 99
PII: 99

Researcher Affiliations

Martino, Nicola A
  • Veterinary Clinics and Animal Productions Unit-Dipartimento dell'Emergenza e Trapianti D'Organo (DETO), Università di Bari Aldo Moro, Str, Prov, Casamassima Km 3°, Valenzano 70010, Bari, Italy. nicmartino@libero.it.
Dell'Aquila, Maria E
    Filioli Uranio, Manuel
      Rutigliano, Lucia
        Nicassio, Michele
          Lacalandra, Giovanni M
            Hinrichs, Katrin

              MeSH Terms

              • Abattoirs
              • Animals
              • Cell Survival
              • Chromatin Assembly and Disassembly
              • Cold Temperature / adverse effects
              • Culture Media
              • Energy Metabolism
              • Female
              • Horses / physiology
              • In Vitro Oocyte Maturation Techniques / veterinary
              • Meiosis
              • Microscopy, Confocal / veterinary
              • Microscopy, Fluorescence / veterinary
              • Mitochondria / metabolism
              • Oocytes / cytology
              • Oocytes / metabolism
              • Oogenesis
              • Oxidation-Reduction
              • Reactive Oxygen Species / metabolism

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