Oxygen tension and oocyte density during in vitro maturation affect the in vitro fertilization of bovine oocytes

Authors

  • Angelo Bertani Giotto Universidade Federal do Pampa
  • Daniela dos Santos Brum Universidade Federal do Pampa
  • Francielli Weber Santos Universidade Federal do Pampa
  • Antonio Carlos Galarça Guimarães Universidade Federal do Pampa
  • Cibele Garcia Moreira Gonçalves Universidade Federal do Pampa
  • Cecilia Urquiza Machado Pavin Universidade Federal do Pampa
  • Natalia Picoli Folchini Universidade Federal do Pampa
  • Aline Barros Moyses Universidade Federal do Pampa
  • Daniele Missio Universidade Federal do Pampa
  • Fábio Gallas Leivas Universidade Federal do Pampa

DOI:

https://doi.org/10.5433/1679-0359.2015v36n6Supl2p4277

Keywords:

Oocyte density, Oxygen tension, ROS production, Superoxide dismutase, GSH.

Abstract

Oocyte maturation is the key factor affecting the fertilization and embryonic development. Factors such as oocyte density and oxygen tension can directly influence the IMV. Thus, the objective of this study was to evaluate the effect of the association of oxygen tensions (5% or 20%) with different oocyte densities (1:10?l or 1:20?l) in the in vitro maturation (IVM) of bovine oocytes on maturation and fertilization rates, ROS production and antioxidant activity. Three experiments were performed with bovine oocytes that were obtained from slaughterhouse ovaries. After selection, the oocytes were randomly distributed in four treatments: 1:10/5%; 1:10/20%; 1:20/5%and 1:20/20% for each experiment. In experiment I, nuclear maturation status and cytoplasmic maturation were evaluated through detection of the first polar body by immunofluorescence and the mitochondrial reorganization assay. In experiment II, ROS production and antioxidant activity were analyzed in oocytes and IVM medium after 24 h of maturation through detection of ROS, reduced glutathione (GSH) and Superoxide dismutase activity by spectrofluorimetric methods. In experiment III, fertilization was evaluated through pronucleus formation, sperm penetration with or without decondensation and polyspermy rates by immunofluorescence. In experiment I, the nuclear maturation and cytoplasmic maturation were similar among treatments (P>0.05). In experiment II, reactive oxygen species in oocytes were elevated in treatments with low oxygen tension which was independent of oocyte density (P<0.05). Additionally, ROS levels in IVM medium were higher in treatments with high oocyte density by volume of medium, which was independent of oxygen tension (P<0.05). In Experiment III, the fertilization and penetration rates were higher in the treatment with 20% oxygen tension and high oocyte density (P<0.05). Furthermore, a high incidence of polyspermy was observed in groups with high oxygen tension and low oocyte density (P<0.05). In conclusion, the results of this study indicate an interaction between oxygen tension and oocyte density, which increases ROS production in certain associations and subsequently influences the rates of in vitro fertilization of bovine oocytes. The improved rates of IVF were obtained when IVM was conducted using 20% oxygen tension and high oocyte density (1:20 ul).

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Author Biographies

Angelo Bertani Giotto, Universidade Federal do Pampa

Discente do Programa de Pós Graduação em Ciência Animal, Universidade Federal do Pampa, UNIPAMPA, Uruguaiana, RS, Brasil.

Daniela dos Santos Brum, Universidade Federal do Pampa

Profa. Associado, UNIPAMPA, Uruguaiana, RS, Brasil.

Francielli Weber Santos, Universidade Federal do Pampa

Profa. Associado, UNIPAMPA, Uruguaiana, RS, Brasil.

Antonio Carlos Galarça Guimarães, Universidade Federal do Pampa

Discente do Programa de Pós Graduação em Ciência Animal, Universidade Federal do Pampa, UNIPAMPA, Uruguaiana, RS, Brasil.

Cibele Garcia Moreira Gonçalves, Universidade Federal do Pampa

Discente do Programa de Pós Graduação em Ciência Animal, Universidade Federal do Pampa, UNIPAMPA, Uruguaiana, RS, Brasil.

Cecilia Urquiza Machado Pavin, Universidade Federal do Pampa

Discente do Programa de Pós Graduação em Ciência Animal, Universidade Federal do Pampa, UNIPAMPA, Uruguaiana, RS, Brasil.

Natalia Picoli Folchini, Universidade Federal do Pampa

Discente do curso de Medicina Veterinária, UNIPAMPA, Uruguaiana, RS, Brasil. E-mail:

Aline Barros Moyses, Universidade Federal do Pampa

Discente do curso de Medicina Veterinária, UNIPAMPA, Uruguaiana, RS, Brasil. E-mail:

Daniele Missio, Universidade Federal do Pampa

Discente do curso de Medicina Veterinária, UNIPAMPA, Uruguaiana, RS, Brasil. E-mail:

Fábio Gallas Leivas, Universidade Federal do Pampa

Prof. Adjunto, UNIPAMPA, Uruguaiana, RS, Brasil.

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Published

2015-12-16

How to Cite

Giotto, A. B., Brum, D. dos S., Santos, F. W., Guimarães, A. C. G., Gonçalves, C. G. M., Pavin, C. U. M., … Leivas, F. G. (2015). Oxygen tension and oocyte density during in vitro maturation affect the in vitro fertilization of bovine oocytes. Semina: Ciências Agrárias, 36(6Supl2), 4277–4288. https://doi.org/10.5433/1679-0359.2015v36n6Supl2p4277

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