Effect of heparin on capacitation/acrosome reaction of equine sperm.
Abstract: The onset of sperm capacitation/acrosome reaction was evaluated using heparin. Equine semen was incubated at 38 degrees C for 4.5 h in culture medium with and without 10 micrograms/mL heparin and with and without 0.1 microM of Ca2+ ionophore. Sperm acrosome reaction was detected using chlortetracycline fluorescence (CTC) and transmission electron microscopy (TEM). The CTC assay provided staining patterns that corresponded with the capacitation/acrosome reaction in other mammalian species (man, mouse, guinea pig). The percentages of incapacitated sperm (PUC), capacitated acrosome-intact sperm (PC), and acrosome-reacted sperm (PAR) were evaluated following incubation times of 0.5 and 4.5 h in heparin-free and heparinized medium, and at 4.5 h only in sperm exposed to Ca2+ ionophore. The CTC assay was highly correlated with TEM for estimation of PAR. At 4.5 h, heparinized medium reduced PUC and increased PC and PAR, in comparison with heparin-free medium. Addition of Ca2+ ionophore to the medium reduced PUC and increased PC and PAR at 4.5 h, as compared with sperm in ionophore-free medium. Incubation time also affected PUC, PC, and PAR in heparin-free and heparinized medium without ionophore. The PUC was greater at 0.5 h than at 4.5 h, and PC and PAR were less at 0.5 h than at 4.5 h. It would appear that the initiation of capacitation/acrosome reaction of equine sperm in vitro is accelerated by heparin.
Publication Date: 1993-11-01 PubMed ID: 8274046DOI: 10.3109/01485019308988400Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article examines the impact of heparin on the process of equine sperm capacitation/acrosome reaction, finding that heparin can accelerate this process.
Research Methodology
- The researchers used equine semen, which was incubated at 38 degrees Celsius for 4.5 hours.
- This semen was then exposed to two types of culture medium, one with and one without 10 micrograms/mL heparin.
- They also carried out tests both with and without 0.1 microM of Ca2+ ionophore, a substance that changes cells’ permeability and allows calcium ions to pass through.
- They employed chlortetracycline fluorescence (CTC) and transmission electron microscopy (TEM) to detect sperm acrosome reaction.
Findings
- The patterns from the CTC assay were found to match the capacitation/acrosome reaction in other mammalian species such as humans, mice, and guinea pigs.
- They calculated the percentages of incapacitated sperm (PUC), capacitated acrosome-intact sperm (PC), and acrosome-reacted sperm (PAR) following incubation times of 0.5 and 4.5 hours in both heparin-free and heparin-infused mediums, and at 4.5 hours with sperm exposed to Ca2+ ionophore.
- Findings indicate a strong correlation between the CTC assay and TEM for estimation of PAR.
- Heparin was found to reduce PUC but increase PC and PAR at the 4.5-hour mark when compared to a heparin-free medium. Similarly, the addition of Ca2+ ionophore also reduced PUC and increased PC and PAR after 4.5 hours.
- Comparisons of incubation time on PUC, PC, and PAR in heparin-free and heparinized medium without ionophore also confirmed a decrease in PUC and increase in PC and PAR from the 0.5-hour to the 4.5 hour mark.
Conclusion
- The findings suggest that heparin accelerates the onset of capacitation/acrosome reaction of equine sperm in vitro.
Cite This Article
APA
Varner DD, Bowen JA, Johnson L.
(1993).
Effect of heparin on capacitation/acrosome reaction of equine sperm.
Arch Androl, 31(3), 199-207.
https://doi.org/10.3109/01485019308988400 Publication
Researcher Affiliations
- Department of Large Animal Medicine, College of Veterinary Medicine, Texas A&M University, College Station 77843-4475.
MeSH Terms
- Acrosome / drug effects
- Acrosome / physiology
- Acrosome / ultrastructure
- Animals
- Chlortetracycline
- Fluorescent Dyes
- Heparin / pharmacology
- Horses
- In Vitro Techniques
- Male
- Sperm Capacitation / drug effects
Citations
This article has been cited 4 times.- Sahoo B, Choudhary RK, Sharma P, Choudhary S, Gupta MK. Significance and Relevance of Spermatozoal RNAs to Male Fertility in Livestock.. Front Genet 2021;12:768196.
- Zapata-Carmona H, Barón L, Zuñiga LM, Díaz ES, Kong M, Drobnis EZ, Sutovsky P, Morales P. The activation of the chymotrypsin-like activity of the proteasome is regulated by soluble adenyl cyclase/cAMP/protein kinase A pathway and required for human sperm capacitation.. Mol Hum Reprod 2019 Oct 28;25(10):587-600.
- Katila T. In vitro evaluation of frozen-thawed stallion semen: a review.. Acta Vet Scand 2001;42(2):199-217.
- Calvete JJ, Mann K, Schäfer W, Sanz L, Reinert M, Nessau S, Raida M, Töpfer-Petersen E. Amino acid sequence of HSP-1, a major protein of stallion seminal plasma: effect of glycosylation on its heparin- and gelatin-binding capabilities.. Biochem J 1995 Sep 1;310 ( Pt 2)(Pt 2):615-22.
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