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Controlled spermatozoa-oocyte interaction improves embryo quality in sheep.

Debora Agata Anzalone1, Luca Palazzese2, Marta Czernik1,2

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Reducing the time sheep oocytes and spermatozoa interact during in vitro fertilization (IVF) significantly improves embryo quality. Shorter incubation times lower polyspermy and oxidative stress, leading to better embryo development and blastocyst formation.

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Area of Science:

  • Reproductive Biology
  • Embryology
  • Animal Science

Background:

  • Current sheep in vitro fertilization (IVF) protocols, using overnight co-incubation of oocytes and spermatozoa, are over 25 years old.
  • These established methods lead to high reactive oxygen species (ROS) concentrations and increased polyspermy due to prolonged gamete interaction.
  • Optimizing IVF requires understanding the precise fertilization time frame to minimize detrimental effects.

Purpose of the Study:

  • To determine the exact time frame for effective sperm penetration and fertilization in sheep oocytes.
  • To compare the outcomes of standard overnight IVF (o/nIVF) with a newly defined short IVF (shIVF) protocol.
  • To evaluate the impact of reduced incubation time on fertilization rates, embryo development, and embryo quality.

Main Methods:

  • Sheep oocytes matured in vitro were co-incubated with spermatozoa in IVF medium.
  • Oocytes were sampled every 30 minutes to identify the fertilization time frame (determined to be 4 hours).
  • Fertilization and embryo development were compared between overnight (16h) and short (4h) incubation groups.

Main Results:

  • Short IVF (shIVF) resulted in significantly lower polyspermic fertilization (6.5%) compared to overnight IVF (o/nIVF) (17.8%).
  • shIVF yielded a higher proportion of 2-cell stage embryos (50.6%) than o/nIVF (34.6%).
  • Embryos developed via shIVF showed improved quality, evidenced by higher blastocyst development rates (29% vs 23.5%) and increased total cell number (TCN).

Conclusions:

  • A reduced spermatozoa-oocyte incubation period (4 hours) enhances sheep embryo production and quality.
  • Shorter incubation minimizes exposure to reactive oxygen species (ROS) and reduces oxidative stress.
  • This optimized IVF approach offers a more efficient and effective method for sheep embryo development.