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Home / Equine Research Bank / PLoS One / In vitro acute exposure to DEHP affects oocyte meiotic matur...
PloS one2011; 6(11); e27452; doi: 10.1371/journal.pone.0027452

In vitro acute exposure to DEHP affects oocyte meiotic maturation, energy and oxidative stress parameters in a large animal model.

Abstract: Phthalates are ubiquitous environmental contaminants because of their use in plastics and other common consumer products. Di-(2-ethylhexyl) phthalate (DEHP) is the most abundant phthalate and it impairs fertility by acting as an endocrine disruptor. The aim of the present study was to analyze the effects of in vitro acute exposure to DEHP on oocyte maturation, energy and oxidative status in the horse, a large animal model. Cumulus cell (CC) apoptosis and oxidative status were also investigated. Cumulus-oocyte complexes from the ovaries of slaughtered mares were cultured in vitro in presence of 0.12, 12 and 1200 µM DEHP. After in vitro maturation (IVM), CCs were removed and evaluated for apoptosis (cytological assessment and TUNEL) and intracellular reactive oxygen species (ROS) levels. Oocytes were evaluated for nuclear chromatin configuration. Matured (Metaphase II stage; MII) oocytes were further evaluated for cytoplasmic energy and oxidative parameters. DEHP significantly inhibited oocyte maturation when added at low doses (0.12 µM; P<0.05). This effect was related to increased CC apoptosis (P<0.001) and reduced ROS levels (P<0.0001). At higher doses (12 and 1200 µM), DEHP induced apoptosis (P<0.0001) and ROS increase (P<0.0001) in CCs without affecting oocyte maturation. In DEHP-exposed MII oocytes, mitochondrial distribution patterns, apparent energy status (MitoTracker fluorescence intensity), intracellular ROS localization and levels, mt/ROS colocalization and total SOD activity did not vary, whereas increased ATP content (P<0.05), possibly of glycolytic origin, was found. Co-treatment with N-Acetyl-Cysteine reversed apoptosis and efficiently scavenged excessive ROS in DEHP-treated CCs without enhancing oocyte maturation. In conclusion, acute in vitro exposure to DEHP inhibits equine oocyte maturation without altering ooplasmic energy and oxidative stress parameters in matured oocytes which retain the potential to be fertilized and develop into embryos even though further studies are necessary to confirm this possibility.
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Publication Date: 2011-11-04 PubMed ID: 22076161PubMed Central: PMC3208636DOI: 10.1371/journal.pone.0027452Google 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 study is basically about understanding the effects of an environmental contaminant, Di-(2-ethylhexyl) phthalate (DEHP), on horse oocytes (female germ cells or eggs) maturation process and consequent energy and stress parameters when exposed to this contaminant in vitro (in a lab setting).

Research Purpose and Methodology

  • The researchers aimed to explore the impact of DEHP, a common environmental pollutant, on the maturation of oocytes (immature egg cells) and the energy and stress parameters within the eggs.
  • The horse was used as a large animal model for this investigation. Oocytes were harvested from horse ovaries via in vitro culture under exposure to varying concentrations of DEHP.
  • Subsequently, the Cumulus cells (cells surrounding the oocyte) were evaluated for signs of apoptosis (programmed cell death), and for levels of reactive oxygen species (ROS), which can indicate stress within the cells.

Research Findings

  • It was found that lower concentrations of DEHP significantly inhibited the maturation of oocytes, a process associated with increased apoptosis in the surrounding cumulus cells and lowered ROS levels.
  • Contrarily, at higher DEHP concentrations, it caused apoptosis and a rise in ROS levels within the CC’s, but it didn’t affect the maturation of oocytes.
  • DEHP exposure in matured oocytes did not alter the distribution of mitochondria, ROS localization and levels, nor the total SOD activity – an antioxidant defense in nearly all living cells exposed to oxygen.
  • However, there was a noted increase in ATP content, the energy currency of the cell. This increase, which may be originated from glycolysis, did not appear to adversely affect the oocytes’ energy status.
  • Co-treatment with N-Acetyl-Cysteine, an antioxidant, was able to reverse cell apoptosis and effectively reduce excessive ROS in DEHP-treated cells, though it did not enhance oocyte maturation.

Research Conclusion

  • The study concluded that acute laboratory exposure to DEHP can inhibit the maturation process of equine oocytes, though matured oocytes’ energy and oxidative stress parameters remain unaltered.
  • According to the results, matured oocytes seem to retain their potential for fertilization and further embryonic development, a possiblity that needs further study to be confirmed. Thus, the implications of DEHP exposure on equine oocyte health are complex and multifaceted, warranting further investigations.

Cite This Article

APA
Ambruosi B, Uranio MF, Sardanelli AM, Pocar P, Martino NA, Paternoster MS, Amati F, Dell'Aquila ME. (2011). In vitro acute exposure to DEHP affects oocyte meiotic maturation, energy and oxidative stress parameters in a large animal model. PLoS One, 6(11), e27452. https://doi.org/10.1371/journal.pone.0027452

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 6
Issue: 11
Pages: e27452
PII: e27452

Researcher Affiliations

Ambruosi, Barbara
  • Department of Animal Production, University of Bari Aldo Moro, Valenzano, Bari, Italy.
Uranio, Manuel Filioli
    Sardanelli, Anna Maria
      Pocar, Paola
        Martino, Nicola Antonio
          Paternoster, Maria Stefania
            Amati, Francesca
              Dell'Aquila, Maria Elena

                MeSH Terms

                • Acetylcysteine / pharmacology
                • Animals
                • Apoptosis / drug effects
                • Cells, Cultured
                • Cumulus Cells / cytology
                • Cumulus Cells / drug effects
                • Cumulus Cells / metabolism
                • Cytoplasm / drug effects
                • Cytoplasm / metabolism
                • Diethylhexyl Phthalate / toxicity
                • Female
                • Free Radical Scavengers / pharmacology
                • Horses
                • Meiosis / drug effects
                • Mitochondria / drug effects
                • Mitochondria / metabolism
                • Models, Animal
                • Oocytes / cytology
                • Oocytes / drug effects
                • Oocytes / metabolism
                • Oogenesis / drug effects
                • Ovarian Follicle / cytology
                • Ovarian Follicle / drug effects
                • Ovarian Follicle / metabolism
                • Oxidation-Reduction
                • Oxidative Stress / drug effects
                • Plasticizers / toxicity
                • Reactive Oxygen Species / metabolism
                • Superoxide Dismutase / metabolism

                Conflict of Interest Statement

                The authors have declared that no competing interests exist.

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                Citations

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