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Follicular fluid hydrogen peroxide and lipid hydroperoxide in bovine antral follicles of various size, atresia, and dominance status

  • Gonadal Physiology and Disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To avoid inducing a state of oxidative stress (OS), assisted reproductive technologies (ART) must maintain a balance of reactive oxygen species (ROS) and antioxidants during the in vitro culture of oocytes. However, oocyte requirements and tolerance thresholds for ROS during in vivo development are still unclear. Previous studies have examined ROS levels in follicular fluid (FF) using pooled samples or according to follicle size. This study sought to examine two OS markers, lipid hydroperoxides (LPO) and hydrogen peroxide (H2O2), in FF of individually sampled follicles from bovine ovary pairs according to follicle size, atresia, and dominance status.

Methods

TUNEL and cleaved Caspase-3 labeling were used to identify apoptotic granulosa cells and determine follicle atresia status. LPO were measured directly for the first time in FF.

Results

Non-atretic follicles and dominant follicles contained more FF H2O2 than atretic follicles and corresponding subordinate follicles, respectively. FF LPO did not vary in relation to atretic status, and no difference existed between dominant and subordinate follicles. However, FF LPO was significantly lower in first subordinate follicles than in the second subordinate follicles from each ovary pair. Neither H2O2 nor LPO levels correlated with follicle size.

Conclusions

These data provide clear evidence that the events of antral folliculogenesis are relevant to ROS dynamics in vivo. Furthermore, such studies will help to optimize in vitro conditions for oocyte culture protocols, particularly when combined with a comparison of oocyte quality with respect to source follicle characteristics.

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Acknowledgments

We thank Richard Single (Department of Mathematics and Statistics, University of Vermont) and Sallie Sheldon for statistical advice; Jim Larrabee and Roger Sandwick (Department of Chemistry and Biochemistry, Middlebury College) for assistance with technical and chemical adjustments to the lipid peroxidation assay; and Tom Sheluga for laboratory support. This project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2011-67016-20041 from the USDA National Institute of Food and Agriculture.

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Authors and Affiliations

  1. Biology Department, Middlebury College, McCardell Bicentennial Hall 346, Middlebury, VT, 05753, USA

    Margo L. Hennet, Hope Y. Yu & Catherine M. H. Combelles

Authors
  1. Margo L. Hennet
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  2. Hope Y. Yu
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  3. Catherine M. H. Combelles
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Correspondence to Catherine M. H. Combelles.

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Capsule

The follicular fluid milieu of dominant and non-atretic follicles is characterized by elevated levels of hydrogen peroxide, while lipid hydroperoxides don’t vary according to follicle size, atresia, and dominance status.

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Hennet, M.L., Yu, H.Y. & Combelles, C.M.H. Follicular fluid hydrogen peroxide and lipid hydroperoxide in bovine antral follicles of various size, atresia, and dominance status. J Assist Reprod Genet 30, 333–340 (2013). https://doi.org/10.1007/s10815-012-9925-5

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  • DOI: https://doi.org/10.1007/s10815-012-9925-5

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