Use of two conventional staining methods to assess the acrosomal status of stallion spermatozoa.
Abstract: The acrosome is a highly specialised region of the spermatozoon that is essential for fertilisation. Defects or dysfunction of this structure have been associated with fertility problems in man and various domestic species including stallions. Current methods of evaluating the acrosome of stallion spermatozoa are time consuming and require specialised equipment, which is cost prohibitive to the average practitioner. Objective: To evaluate 2 conventional stains (Dip Quick and Spermac) and determine their usefulness in assessing acrosome integrity in stallions as compared with specific acrosomal labelling with a fluorescein-conjugated lectin - a method that has been validated for acrosome status evaluation in stallions. Methods: In vivo experimental design. Methods: Semen from 6 mature Miniature horse stallions of known fertility was collected on 5 separate occasions. To increase the number of reacted acrosomes, portions of each ejaculate were incubated with the calcium ionophore, A23187. Ejaculates were divided and semen samples were processed according to recommendations for fluorescein-conjugated peanut lectin, Pisum sativum agglutin, Dip Quick, and Spermac staining methods. Slides were evaluated independently by 2 separate investigators. Spermatozoa were classified as having intact, reacting, reacted or defective acrosomes. Results: All parameters obtained by both investigators, using all 3 staining methods were highly correlated (P0.05) between investigators or staining method for the percentages of intact or reacted acrosomes. However, there was a significant difference between investigators and staining methods for determining reacting acrosome percentages (P<0.05). Conclusions: Dip Quick and Spermac stains are useful for determining intact vs. reacted acrosomes for stallion spermatozoa.
© 2013 EVJ Ltd.
Publication Date: 2013-11-20 PubMed ID: 24004290DOI: 10.1111/evj.12179Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research examines the effectiveness of two conventional staining methods, Dip Quick and Spermac, in evaluating the acrosome status of stallion spermatozoa, in comparison with a validated fluorescein-conjugated lectin method. The findings suggest these staining methods are useful for differentiating between intact and reacted acrosomes in stallion spermatozoa.
Objective of the Research
- The study sets out to explore the efficiency of two commonly used stains – Dip Quick and Spermac in evaluating the integrity of acrosome, a vital part of the spermatozoon, in stallions.
- The study also contrasts the practicality and effectiveness of these stains with a fluorescein-conjugated lectin method that is validated for stallion acrosome status analysis.
Methodology
- The methodology involved in vivo experimental design.
- Semen samples were collected from six mature Miniature horse stallions of proven fertility on five separate instances.
- Portions of each ejaculate were incubated with the calcium ionophore, A23187, to amplify the number of reacted acrosomes.
- Ejaculates were divided and processed in accordance with the recommended protocols for fluorescein-conjugated peanut lectin, Pisum sativum agglutin, Dip Quick, and Spermac staining procedures.
- The samples were evaluated independently by two different investigators. They classified spermatozoa based on the acrosome status – intact, reacting, reacted, or defective.
Results
- All parameters obtained by both investigators using all three staining procedures were highly correlated.
- There was no significant difference between the investigators or between the staining methods when examining the percentages of intact or reacted acrosomes (P>0.05).
- However, a significant difference was found between investigators and staining methods when determining the percentages of reacting acrosomes (P<0.05).
Conclusion
- The findings concluded that Dip Quick and Spermac stains are useful in categorising stallion spermatozoa with intact versus reacted acrosomes.
Cite This Article
APA
Runcan EE, Pozor MA, Zambrano GL, Benson S, Macpherson ML.
(2013).
Use of two conventional staining methods to assess the acrosomal status of stallion spermatozoa.
Equine Vet J, 46(4), 503-506.
https://doi.org/10.1111/evj.12179 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine University of Florida, Gainesville, USA.
MeSH Terms
- Acrosome / physiology
- Animals
- Horses / physiology
- Male
- Semen Analysis / veterinary
- Staining and Labeling / methods
- Staining and Labeling / veterinary
Citations
This article has been cited 1 times.- Farias LB, da Cunha Barreto-Vianna AR, de Mello MD, Dos Santos AL, da Fonte Ramos C, Fontoura P. Comparison of Diff-Quick and Spermac Staining Methods for Sperm Morphology Evaluation.. J Reprod Infertil 2023 Jul-Sep;24(3):166-170.
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