Adaptation of the hypoosmotic swelling test to assess functional integrity of stallion spermatozoal plasma membranes.
Abstract: Hypoosmotic swelling (HOS) is used for assessing plasma membrane function and fertilizing capacity of human spermatozoa. However, HOS solutions and methodologies have not been evaluated specifically for assessing stallion spermatozoa. The objective of this study was to identify a HOS solution and assay conditions specifically for stallions that would maximize spermatozoal plasma membrane swelling. The HOS solutions and assay conditions, including incubation time (15 to 180 min), temperature (25 degrees vs 37 degrees C), and total number of cells examined (100, 200 or 500) were evaluated. Assay consistency, accuracy, reliability and repeatability also were determined. Maximum spermatozoal plasma membrane swelling was observed in a 100 mosmol sucrose solution (P < 0.001). Incubation time (P = 0.67), temperature (P = 0.70) and total number of spermatozoa examined (P = 0.38 and P = 0.24 for 100 vs 200 and 100 vs 500, respectively) did not influence percent of HOS positive spermatozoa observed. A high degree of assay accuracy was indicated when a correlation of r = 0.998 was obtained between the HOS positive spermatozoa observed and expected when known amounts of heat-treated spermatozoa, unable to undergo swelling, were added to untreated spermatozoa. Assay consistency was demonstrated, as reflected by a mean coefficient of variation of 0.073 for 4 stallions. Also, the coefficient of variation from 2 analyses of variance was 0.168 and 0.096, indicating reasonably good assay reliability; estimates of repeatability from the same analyses were 0.794 and 0.968. The HOS test adapted to stallion spermatozoa in this study is a simple, highly accurate and consistent assay with good reliability and repeatability. Results observed under the conditions evaluated also permit some flexibility in adapting this assay to individual laboratory and practice settings for evaluating stallion spermatozoal plasma membranes.
Publication Date: 2001-04-09 PubMed ID: 11291907DOI: 10.1016/s0093-691x(01)00461-7Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The abstract discusses a study that aimed to adapt the hypoosmotic swelling (HOS) test to evaluate the plasma membrane functionality in stallion sperm cells. The research proved the test’s effectiveness, demonstrating accuracy, consistency, and reliability, with scope for customization to individual laboratory conditions.
Objective and Methodology
- The primary pursuit of this study was to find an appropriate hypoosmotic swelling (HOS) solution and to establish assay conditions for evaluating the plasma membrane function of stallion sperm.
- The researchers tested various HOS solutions and conditions, which included different incubation durations (ranging from 15 to 180 minutes), temperatures (25 degrees Celsius or 37 degrees Celsius), and the total number of cells scrutinized (ranging between 100 to 500).
- Furthermore, the study aimed to determine the level of consistency, accuracy, reliability, and repeatability of the test.
Findings
- The study observed that the maximum swelling of the sperm cell plasma membrane took place in a 100 mosmol sucrose solution.
- The researchers found that the incubation period, temperature, and the total quantity of sperm cells examined did not significantly impact the percentage of HOS positive sperm cells observed.
- The accuracy of the assay was validated since a high correlation (r = 0.998) existed between the expected and observed HOS positive sperm when known amounts of heat-treated sperm (which couldn’t undergo swelling) were added to untreated sperm cells.
Consistency, Reliability, and Repeatability
- The consistency of the assay was demonstrated by a mean coefficient variation of 0.073 for four stallions.
- The coefficients of variations presented were 0.168 and 0.096, illustrating good assay reliability.
- Repeatability measurements from the same analyses were found to be 0.794 and 0.968, proving the test’s repeatability and consistency.
Conclusion
- The research concludes that the HOS test, when adapted for stallion sperm cells, is a simple, highly accurate, and consistent test with good reliability and repeatability.
- It is noteworthy that the conditions of the assay tested also allow room for some flexibility, enabling adaptation to individual lab and practice settings for evaluating stallion sperm cells’ plasma membranes.
Cite This Article
APA
Nie GJ, Wenzel JG.
(2001).
Adaptation of the hypoosmotic swelling test to assess functional integrity of stallion spermatozoal plasma membranes.
Theriogenology, 55(4), 1005-1018.
https://doi.org/10.1016/s0093-691x(01)00461-7 Publication
Researcher Affiliations
- Department of Large Animal Surgery & Medicine, College of Veterinary Medicine Auburn University, Alabama 36849-5522, USA.
MeSH Terms
- Animals
- Cell Membrane / physiology
- Cell Size
- Horses / physiology
- Hypotonic Solutions
- Male
- Reproducibility of Results
- Sensitivity and Specificity
- Sperm Count
- Spermatozoa / ultrastructure
- Temperature
- Time Factors
Citations
This article has been cited 3 times.- Alamaary MS, Haron AW, Ali M, Hiew MWH, Adamu L, Peter ID. Effects of four extenders on the quality of frozen semen in Arabian stallions.. Vet World 2019 Jan;12(1):34-40.
- Absalan F, Ghannadi A, Zabihi A. The effects of different doses of ketamine on quality of normal ejaculated sperm.. Int J Fertil Steril 2014 Jul;8(2):207-14.
- Katila T. In vitro evaluation of frozen-thawed stallion semen: a review.. Acta Vet Scand 2001;42(2):199-217.
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