Effect of capacitation of stallion sperm with polyvinylalcohol or bovine serum albumin on penetration of bovine zona-free or partially zona-removed equine oocytes.
Abstract: Experiments were conducted to study effects of macromolecules on stallion sperm capacitation and fertilization as determined by penetration of bovine zona-free and equine partially zona-removed oocytes. Stallion sperm were capacitated in TYH medium (modified Krebs-Ringer bicarbonate) supplemented with either 1 mg/mL of polyvinylalcohol (PVA) or 4 mg/mL of BSA. Capacitation was induced with 8 bromoadenosine cyclic monophosphate (8BrcAMP; 0.5 mM) alone or in combination with 0.1 microM of ionomycin. Intraspecies gametes were co-incubated in TYH/PVA or TYH/BSA for 18 to 20 h. For zona-free bovine oocytes, penetration rate (35%) with the combination of 8BrcAMP and ionomycin in PVA-containing medium was higher (P < 0.05) than any treatment in BSA-containing medium (5 to 6%). A similar study was conducted using equine oocytes with partially removed zonae. Sperm capacitated and used for in vitro fertilization (IVF) in PVA-containing medium had higher penetration rates (P < 0.01) than sperm in BSA-containing medium (54 vs. 11%). The effect of equine preovulatory follicular fluid on bovine oocyte penetration was assessed. Bovine oocytes were matured in tissue culture medium-199 with 0, 20, 50, or 100% equine preovulatory follicular fluid, and 1 IU/mL of equine chorionic gonadotropin. Stallion sperm were treated with 8BrcAMP + ionomycin in PVA- or BSA-containing media. The penetration rates of bovine zona-free oocytes by stallion sperm were again higher with PVA (47%) than BSA (18%; P < 0.01). Penetration rates of oocytes matured in 100% follicular fluid were higher (P < 0.05) than for oocytes matured with 0% follicular fluid. The effects of equine follicular fluid and PVA/BSA during sperm capacitation on standard bovine IVF were examined. Culture of bovine oocytes with equine follicular fluid did not affect oocyte maturation or penetration rates after IVF. Bovine sperm capacitated with heparin in PVA-containing medium yielded lower (P < 0.05) fertilization rates than those capacitated in BSA-containing medium when incubated with both zona-intact and zona-free bovine oocytes. In summary, PVA was superior to BSA for ionophore-induced capacitation of equine sperm for penetration of zona-free bovine oocytes or partially zona-removed equine oocytes, but not for standard bovine IVF with bovine sperm. Zona-free bovine oocytes may be useful for assaying in vitro capacitation and fertilization of stallion sperm.
Publication Date: 2003-08-21 PubMed ID: 12926790DOI: 10.2527/2003.8182080xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research studied how certain macromolecules affect the capacitation (preparation for fertilization) and fertilization of stallion sperm cells. Using two different substances, polyvinylalcohol or bovine serum albumin, it was found that polyvinylalcohol was a superior supplement for this process in certain situations, but not for standard bovine in vitro fertilization using bovine sperm.
Methods
- Stallion sperm cells were prepared in a specialized medium, supplemented with either 1 mg/mL of polyvinylalcohol (PVA) or 4 mg/mL of bovine serum albumin (BSA).
- Capacitation was induced with a compound called 8 bromoadenosine cyclic monophosphate (8BrcAMP), either alone or together with a substance called ionomycin.
- These prepared sperm cells were incubated with corresponding species oocytes (female reproductive cells) in either PVA or BSA solutions for around 18 to 20 hours.
Results
- For bovine oocytes that had no zona (a protective layer around the oocyte), the combination of 8BrcAMP and ionomycin in a PVA medium resulted in a higher penetration rate than any treatment involving BSA. This suggests that PVA was more effective at promoting capacitation and improving the chances of fertilization.
- Similar results were observed when equine oocytes with partially removed zonae were used. Sperm prepared in PVA had higher penetration rates than those prepared in BSA.
- The effects of equine preovulatory follicular fluid (fluid containing mature oocytes right before ovulation) on the penetration of bovine oocytes were also observed. Higher penetration rates were achieved with PVA, and penetration rates also increased when oocytes were matured in 100% follicular fluid.
- However, while PVA seemed to be more effective in these scenarios, its performance in standard bovine IVF procedures using bovine sperm was less effective. Bovine sperm capacitated in a PVA medium produced lower fertilization rates than the ones prepared in BSA.
Conclusion
- The study concluded that while PVA was generally superior to BSA for promoting capacitation and improving penetration rates in some scenarios, it did not prove effective in standard bovine in vitro fertilization using bovine sperm.
- This research indicates that bovine oocytes with no zona might be a useful tool to study how different conditions affect the in vitro capacitation and fertilization process of stallion sperm.
Cite This Article
APA
Choi YH, Landim-Alvarenga FC, Seidel GE, Squires EL.
(2003).
Effect of capacitation of stallion sperm with polyvinylalcohol or bovine serum albumin on penetration of bovine zona-free or partially zona-removed equine oocytes.
J Anim Sci, 81(8), 2080-2087.
https://doi.org/10.2527/2003.8182080x Publication
Researcher Affiliations
- Department of Biomedical Sciences, Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523, USA.
MeSH Terms
- 8-Bromo Cyclic Adenosine Monophosphate / pharmacology
- Animals
- Cattle
- Culture Techniques / veterinary
- Female
- Fertilization in Vitro / veterinary
- Follicular Fluid / physiology
- Horses / physiology
- Male
- Oocytes
- Polyvinyl Alcohol / pharmacology
- Serum Albumin, Bovine / pharmacology
- Sperm Capacitation / drug effects
- Sperm Capacitation / physiology
- Sperm-Ovum Interactions / drug effects
- Zona Pellucida / physiology
Citations
This article has been cited 4 times.- Felix MR, Turner RM, Dobbie T, Hinrichs K. Successful in vitro fertilization in the horse: production of blastocysts and birth of foals after prolonged sperm incubation for capacitation†. Biol Reprod 2022 Dec 10;107(6):1551-1564.
- Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE. Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status. Animals (Basel) 2021 Jan 3;11(1).
- Liu L, Nutter LM, Law N, McKerlie C. Sperm freezing and in vitro fertilization in three substrains of C57BL/6 mice. J Am Assoc Lab Anim Sci 2009 Jan;48(1):39-43.
- Xiong J, Xiong C, Tian Y, Hu L, Wei H. Effects of murine cytomegalovirus infection on sperm viability in mice. J Huazhong Univ Sci Technolog Med Sci 2006;26(1):130-2.
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