Assessment of Pisum sativum agglutinin in identifying acrosomal damage in stallion spermatozoa.
Abstract: The use of fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) was evaluated for its ability to distinguish acrosome-intact from acrosome-damaged stallion spermatozoa. Incubation of fresh (acrosome-intact) and frozen-thawed (acrosome-damaged) spermatozoa with FITC-PSA resulted in acrosome-intact spermatozoa that exhibited no fluorescence, while acrosome-damaged spermatozoa exhibited fluorescent staining over the rostral portion of the head and equatorial segment. Experiments using mixtures of various ratios of acrosome-intact and acrosome-damaged spermatozoa determined the precision (intrasample coefficient of variation), and linearity (increased percentage of spermatozoa with PSA binding, with increased percentage of frozen-thawed spermatozoa in a sample) of FITC-PSA binding. The binding of FITC-PSA increased in samples as the portion of frozen-thawed (acrosome-damaged) to fresh (acrosome-intact) spermatozoa increased. A positive correlation existed (r = 0.98, P less than 0.05) between the percentage of FITC-PSA bound sperm and the proportion of damaged spermatozoa added to a sample. Location of PSA lectin binding on acrosome-damaged spermatozoa, determined by electron microscopy using gold-conjugated PSA, was to components of the outer acrosomal membrane and acrosomal matrix. These results demonstrate that FITC-PSA binding may be useful in determining acrosomal integrity of fresh and frozen-thawed stallion spermatozoa.
Publication Date: 1992-05-01 PubMed ID: 1515146DOI: 10.1002/mrd.1080320105Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study investigates the effectiveness of a substance called FITC-PSA in distinguishing between healthy and damaged sperm cells in horses.
About the Research
- The research explores the function of fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA), a type of substance derived from peas, and its capacity to differentiate between acrosome-intact (healthy) and acrosome-damaged (damaged) sperm in stallions (male horses).
- The study sought to understand if FITC-PSA could clearly delineate between these two states of cells by its adherence or lack thereof. If successful, it could serve as a tool for determining the viability and quality of sperm, which is useful in breeding and veterinary medicine.
Methods and Findings
- The researchers subjected fresh (healthy) and frozen-thawed (damaged) spermatozoa to FITC-PSA. They observed that acrosome-intact or fit spermatozoa exhibited no fluorescence (or glow) when exposed to FITC-PSA. On the contrary, acrosome-damaged spermatozoa manifested fluorescent staining on the rostral (front) part of the head and equatorial segment, thus marking them out.
- They conducted experiments with varying ratios of healthy and damaged sperm to verify the reliability (precision and linearity) of FITC-PSA’s binding to the cells. Reception of FITC-PSA was noted to increase as the proportion of frozen-thawed (damaged) to fresh (healthy) sperm increased, which further helped with the differentiation.
- Additionally, a strongly positive correlation (r = 0.98, statistically significant P value less than 0.05) was seen between the percentage of FITC-PSA-bound sperm and the ratio of damaged spermatozoa in a sample.
- Meticulous examination through electron microscopy noted that the FITC-PSA quite accurately clung to the outer acrosomal membrane and acrosomal matrix in the case of acrosome-damaged spermatozoa.
Implications
- These findings indicate that FITC-PSA has the potential to determine the health (acrosomal integrity) of fresh as well as frozen stallion spermatozoa.
- The ability to identify and quantify damaged sperm in relation to healthy ones could assist in developing strategies for animal reproduction, particularly in horse breeding.
- This method, being relatively simple and effective, could become a regular feature in veterinary diagnostics pertaining to reproduction.
Cite This Article
APA
Farlin ME, Jasko DJ, Graham JK, Squires EL.
(1992).
Assessment of Pisum sativum agglutinin in identifying acrosomal damage in stallion spermatozoa.
Mol Reprod Dev, 32(1), 23-27.
https://doi.org/10.1002/mrd.1080320105 Publication
Researcher Affiliations
- Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523.
MeSH Terms
- Acrosome / ultrastructure
- Animals
- Cryopreservation
- Fluorescein-5-isothiocyanate
- Horses / physiology
- Lectins / metabolism
- Male
- Membrane Glycoproteins / metabolism
- Microscopy, Electron
- Microscopy, Fluorescence
- Plant Lectins
- Semen Preservation
Citations
This article has been cited 1 times.- Katila T. In vitro evaluation of frozen-thawed stallion semen: a review. Acta Vet Scand 2001;42(2):199-217.
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