Subjecting horse spermatozoa to hypoosmotic incubation: effects of ouabain.
Abstract: Although hypoosmotic tests are widely used to assess spermatozoal quality in different species, they have not been used extensively in the stallion. Moreover, the role of the Na (+)K (+), ouabain sensitive-ATP-ase in the response of equine sperm to hypoosmotic shock is not well understood. This study tests two hypotheses: 1) that equine spermatozoa will respond to a hypoosmotic medium by swelling of the tail, and 2) that addition of ouabain will increase the percentage of swollen sperm tails. Ejaculates from 3 stallions were collected with an artificial vagina and diluted in Kenney's medium (Time = 0). Aliquots were randomly selected to be incubated in an isoosmotic (297 mOsm) or different hypoosmotic media that were composed of citrate or of citrate wïth fructose. The osmolarity of the hypoosmotic media with citrate ranged from 18 to 96 mOsm, and the medium composed of citrate plus fructose (HOS medium) was of 153 mOsm. Moreover, aliquots of spermatozoa pretreated with ouabain were added to the isoosmotic medium and also to the HOS and the 96 mOsm citrate medium (ORT medium). Incubation of equine sperm in the hypoosmotic media resulted in a time- and osmolarity-dependent swelling of the sperm tail, reaching maximum values after incubation for 20-30 min in both the HOS and ORT media. Ouabain induced a dose-dependent effect on swollen tails and viability in fresh semen and also affected some parameters related to motility. Ouabain also increased the swelling response in a hypoosmotic medium although viability decreased. The percentage of swollen tails after incubation in ORT and HOS media snowed significant correlations to viability, altered acrosomes and total motility, but not to other parameters of horse semen analysis. Our results suggest that hypoosmotic tests could be used to improve standard horse semen analysis. Additionally, Na (+)K (+)-ATP-ase activity could be related to the response against hypoosmotic shock of horse spermatozoa.
Publication Date: 1997-02-01 PubMed ID: 16728027DOI: 10.1016/s0093-691x(97)00033-2Google Scholar: Lookup
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- Journal Article
Summary
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This research is about exploring the effects of ouabain and hypoosmotic conditions on horse spermatozoa, suggesting that such conditions could be useful in assessing sperm quality and could have implications for understanding the role of Na(+)K(+)-ATP-ase activity in sperm.
Research Objectives & Hypotheses
- The study aimed to investigate how horse sperm reacts to a hypoosmotic environment, particularly in regards to tail swelling, and how ouabain—an inhibitor of the sodium-potassium pump (Na(+)K(+)-ATP-ase)—affects this reaction.
- The researchers hypothesized that horse sperm would respond to a hypoosmotic medium by swelling of the tail and that the addition of ouabain would increase the percentage of swollen sperm tails.
Methodology
- Ejaculates from three stallions were collected and diluted, then a part of each sample was incubated in either an isoosmotic medium, or in hypoosmotic media of various osmolarity that was composed of citrate alone, or citrate with fructose.
- Semen samples were also pretreated with ouabain before being added to the media.
- The researchers monitored changes in sperm, such as tail swelling and overall viability, over time in response to the different conditions.
Findings
- The study found that exposing horse sperm to hypoosmotic media caused a time- and osmolarity-dependent swelling of the sperm tail.
- Maximum tail swelling was observed after approximately 20-30 minutes of incubation in both the hypoosmotic citrate medium and the citrate plus fructose medium.
- Ouabain application has been shown to exacerbate the swelling effect but also decreased sperm viability and affected some motility-related parameters.
Conclusions & Implications
- The research suggests that hypoosmotic tests could be useful tools for improving the standard analysis of horse semen.
- Furthermore, the response of horse sperm to hypoosmotic shock seems related to the activity of Na(+)K(+)-ATP-ase, suggesting a possible area for further study in sperm biology and fertility.
Cite This Article
APA
de la Cueva FI, Rigau T, Bonet S, Miró J, Briz M, Rodríguez-Gil JE.
(1997).
Subjecting horse spermatozoa to hypoosmotic incubation: effects of ouabain.
Theriogenology, 47(3), 765-784.
https://doi.org/10.1016/s0093-691x(97)00033-2 Publication
Researcher Affiliations
- Reproduction Unit, Department of Animal Pathology and Production, School of Veterinary Medicine, Autonomous University of Barcelona, E-08193 Bellaterra, Spain.
Citations
This article has been cited 2 times.- McCarthy MJ, Baumber J, Kass PH, Meyers SA. Osmotic stress induces oxidative cell damage to rhesus macaque spermatozoa.. Biol Reprod 2010 Mar;82(3):644-51.
- Corral-Baqués MI, Rigau T, Rivera M, Rodríguez JE, Rigau J. Effect of 655-nm diode laser on dog sperm motility.. Lasers Med Sci 2005;20(1):28-34.
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