In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer.
Abstract: Three experiments were conducted to evaluate the effect of oocyte and sperm treatments on rates of in vitro fertilization (IVF) in the horse and to determine the capacity of in vitro-matured horse oocytes to be fertilized in vivo. There was no effect of duration of oocyte maturation (24 vs. 42 h) or calcium ionophore concentration during sperm capacitation (3 microM vs. 7.14 microM) on in vitro fertilization rates. Oocytes matured in 100% follicular fluid had significantly higher fertilization (13% to 24%) than did oocytes matured in maturation medium or in 20% follicular fluid (0% to 12%; P < 0.05). There was no significant difference in fertilization rate among 3 sperm treatments utilizing 7.14 microM calcium ionophore (12% to 21%). Of in vitro-matured oocytes recovered 40-44 h after transfer to the oviducts of inseminated mares, 77% showed normal fertilization (2 pronuclei to normal cleavage). Cleavage to 2 or more cells was seen in 22% of oocytes matured in follicular fluid and 63% of oocytes matured in maturation medium; this difference was significant (P < 0.05). We conclude that in vitro-matured horse oocytes are capable of being fertilized at high rates in the appropriate environment and that in vitro maturation of oocytes in follicular fluid increases fertilization rate in vitro but reduces embryo development after fertilization in vivo. Further work is needed to determine the optimum environment for sperm capacitation and IVF in the horse.
Publication Date: 2002-06-25 PubMed ID: 12080025DOI: 10.1095/biolreprod67.1.256Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research investigated the impact of various oocyte and sperm treatments on horse in vitro fertilization (IVF), and the ability of in vitro-matured horse oocytes to be fertilized in vivo. It found that the environment in which oocyte matures significantly affects fertilization rates but duration of maturation and sperm treatment have no effect.
Study Design and Experiments
- Three different experiments were conducted to understand how oocyte and sperm treatments influence IVF in horses. The ultimate goal was to see whether in vitro-matured horse oocytes can be fertilized in vivo.
- The study compared two durations of oocyte maturation: 24 hours and 42 hours. Two concentrations of calcium ionophore during sperm capacitation were also compared: 3 microM and 7.14 microM. The rate of IVF was not significantly affected by either of these variables.
Follicular Fluid versus Maturation Medium
- Oocytes that matured in 100% follicular fluid had a significantly higher fertilization rate (13 to 24%) compared to those matured in either a maturation medium or in 20% follicular fluid (0 to 12%).
- However, beyond fertilization, the study also found a significant difference in embryonic development between the two maturation environments. A significantly higher percentage of oocytes matured in maturation medium cleaved into two or more cells (63%) than those matured in follicular fluid (22%).
Sperm Treatment and In Vivo Fertilization
- Three sperm treatments using 7.14 microM calcium ionophore were applied. There was no significant difference in fertilization rate among these treatments (12 to 21%).
- The study also included an in vivo verification experiment, where in vitro-matured oocytes were transferred to the oviducts of inseminated mares. Of the oocytes recovered 40-44 hours after the procedure, 77% showed normal fertilization (displaying 2 pronuclei to normal cleavage).
Conclusion
- The research concluded that horse oocytes matured in vitro are capable of being fertilized at high rates, given the right environment.
- Natural maturation in follicular fluid boosted the in vitro fertilization rate. However, this appears to limit embryo development post-fertilization in a natural in vivo setting.
- The paper highlights the need for additional research to determine the best environment for sperm capacitation and IVF in horses.
Cite This Article
APA
Hinrichs K, Love CC, Brinsko SP, Choi YH, Varner DD.
(2002).
In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer.
Biol Reprod, 67(1), 256-262.
https://doi.org/10.1095/biolreprod67.1.256 Publication
Researcher Affiliations
- Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843-4466, USA. khinrichs@cvm.tamu.edu
MeSH Terms
- Animals
- Calcium / pharmacology
- Culture Media
- Fallopian Tubes / physiology
- Female
- Fertilization / physiology
- Fertilization in Vitro
- Fluorescent Dyes
- Horses / physiology
- In Vitro Techniques
- Ionophores / pharmacology
- Male
- Oocytes / physiology
- Sperm Motility / physiology
- Spermatozoa / drug effects
Citations
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