In vitro induction of acrosome reactions in stallion spermatozoa by heparin and A23187.
Abstract: The ability of the glycosaminoglycan, heparin, and the calcium ionophore, A23187, to induce acrosome reaction in equine spermatozoa was assessed using semen from 3 warmblood stallions of known high fertility. After collection of semen, the spermatozoa were washed and incubated in vitro with heparin or A23187. Incubation periods were 0, 4, 6 or 8 h with 0, 1, 10 or 100 microg/ml heparin or 0, 10, 30 or 60 min with 0, 0.01, 0.1, 1 or 10 microM A23187, respectively. Acrosome reactions were determined by staining the spermatozoa with naphthol yellow S plus erythrosin B, and sperm viability was assessed by eosin B-nigrosin staining. Both stains were evaluated under bright-field illumination at x 1000 magnification. Maximal percentages of acrosome reactions were found to occur after incubation for 4 h with 100 microg/ml heparin (71.8 +/- 3.5 % acrosome-reacted spermatozoa compared with 18.7 +/- 1.1 % in control spermatozoa; P < 0.001) or with either 1 or 10 microM A23187 for 60 min (44.0 +/- 6.2 and 45.3 +/- 5.0 % acrosome reacted spermatozoa, respectively, compared with 17.8 +/- 1.5 % in the controls, both P < 0.01). Maximal responses to these conditions varied significantly between stallions (P < 0.01). These results indicate that acrosome reaction can be successfully induced in vitro in stallion spermatozoa with both heparin and A23187, a possible basis for the laboratory prediction of fertility in this species.
Publication Date: 1996-04-15 PubMed ID: 16727876DOI: 10.1016/0093-691x(96)00075-1Google Scholar: Lookup
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- Journal Article
Summary
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This research explores the use of the glycosaminoglycan heparin and the calcium ionophore A23187 to stimulate an acrosome reaction in equine spermatozoa, which can help to predict fertility in stallions.
Objective
The study aims to measure the effect of heparin and A23187 on inducing acrosome reaction in equine spermatozoa. The purpose of detecting these reactions is to potentially use them as an in-lab method for predicting stallion fertility.
Methodology
- The research utilised sperm samples from three warmblood stallions known for their high fertility.
- The collected semen was thoroughly cleaned before the start of the experiment.
- The sperm cells were then incubated, in lateral conditions, with either heparin or A23187.
- In heparin’s case, the incubation lasted 0, 4, 6 or 8 hours and measure 0, 1, 10 or 100 micrograms per milliliter.
- For the calcium ionophore A23187, incubation ranged from 0 to 60 minutes with doses varying from 0 to 10 micromolar.
- Any resulting acrosome reactions were tracked through staining with naphthol yellow S plus erythrosin B.
- Sperm viability was assessed by way of eosin B-nigrosin staining.
- Both stains were examined under bright-field illumination at a 1000x magnification.
Findings
- The highest percentage of induced acrosome reactions was found after a 4-hour incubation with 100 microg/ml of heparin or after a 60-minute incubation with either 1 or 10 microM A23187.
- Compared to controls, these conditions led to a significant increase in acrosome reactions. For instance, the percentage of reacted spermatozoa went up to 71.8% in the case of heparin exposure, compared with 18.7% in control sperm cells.
- The reactions under these conditions varied significantly amongst the stallions.
Conclusion
The study concludes that stimulating acrosome reactions in stallion spermatozoa in laboratory conditions using either glycosaminoglycan heparin or the calcium ionophore A23187 is feasible. These findings are important as they can potentially provide a means for predicting stallion fertility.
Cite This Article
APA
Christensen P, Whitfield CH, Parkinson TJ.
(1996).
In vitro induction of acrosome reactions in stallion spermatozoa by heparin and A23187.
Theriogenology, 45(6), 1201-1210.
https://doi.org/10.1016/0093-691x(96)00075-1 Publication
Researcher Affiliations
- Hampshire Cattle Breeders Society, Ltd., Beechen Lane, Lyndhurst, Hampshire, SO43 7NN, UK.
Citations
This article has been cited 1 times.- McPartlin LA, Visconti PE, Bedford-Guaus SJ. Guanine-nucleotide exchange factors (RAPGEF3/RAPGEF4) induce sperm membrane depolarization and acrosomal exocytosis in capacitated stallion sperm. Biol Reprod 2011 Jul;85(1):179-88.
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