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Home / Equine Research Bank / Theriogenology / Dynamics of sperm DNA fragmentation in domestic animals II. ...
Theriogenology2007; 68(9); 1240-1250; doi: 10.1016/j.theriogenology.2007.08.029

Dynamics of sperm DNA fragmentation in domestic animals II. The stallion.

Abstract: The mixed success of equine artificial insemination programs using chilled and frozen-thawed semen is most likely associated with the variable response of the sperm cell to the preservation process and the fact that stallions are not selected on the basis of reproductive performance. We propose that the traditional indicators of sperm viability do not fully account for male factor infertility in the stallion and that knowledge of sperm DNA damage in the original semen sample and during semen processing may provide a more informed explanation of an individual stallion's reproductive potential. This study reports on the validation of a sperm DNA fragmentation test based on the sperm chromatin dispersion test (SCD) for stallion spermatozoa and on its application to semen that was chilled (4 degrees C; n=10) or frozen-thawed (n=13). Semen samples were collected by artificial vagina and the proportion of sperm with fragmented DNA determined. Seminal plasma was then removed by centrifugation and the sperm pellet re-suspended in commercial extenders prior to being chilled or cryopreserved using standard industry protocols. Chilled semen was cooled slowly to 4 degrees C and stored for 1h before commencing the analysis; cryopreserved semen was thawed and immediately analyzed. Following chilling or cryopreservation, the semen samples were incubated at 37 degrees C and analyzed for SCD after 0, 4, 6, 24 and 48 h storage. The results of this investigation revealed that there was no significant difference in the sperm DNA fragmentation index (sDFI) of sperm evaluated initially after collection compared to those tested immediately after chilling or cryopreservation. However, within 1h of incubation at 37 degrees C, both chilled and frozen-thawed spermatozoa showed a significant increase in the proportion of sDFI; after 6h the sDFI had increased to over 50% and by 48 h, almost 100% of the sperm showed DNA damage. While the sDFI of individual stallions at equivalent times of incubation was variable, an analysis of the rate of change of sDFI revealed no difference between stallions or the way in which the semen was preserved. In terms of sperm DNA fragmentation dynamics, the highest intensity of sperm DNA damage occurred in the first 6h of incubation. We suggest that the SCD test can be used as a routine assessment tool for the development and refinement of preservation protocols designed to reduce stallion sperm DNA damage.
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Publication Date: 2007-10-04 PubMed ID: 17919715DOI: 10.1016/j.theriogenology.2007.08.029Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research study is about the viability of sperm cells in stallions throughout the preservation process. Using a newly developed testing method, the researchers investigated the damage that occurs in the sperm DNA during chilling and freeze-thawing process, which can impact the success of artificial insemination programs.

Research Subject and Method

  • The study focuses on the viability of stallion’s sperm cells during cooling and freeze-thawing process used for artificial insemination programs.
  • To examine this, a sperm DNA fragmentation test known as the sperm chromatin dispersion test (SCD) was used. It was rigorously tested on stallion spermatozoa samples collected by artificial vagina.
  • The semen was then processed – the seminal plasma was removed, and the sperm pellet was re-suspended in commercial extenders. The samples were either chilled or cryopreserved following established industry protocols.

Sperm Samples and Analysis

  • Chilled semen was slowly brought to 4 degrees C and analyzed after 0, 4, 6, 24, and 48 hours. With cryopreserved semen, the samples were thawed and the analysis started immediately. Between the tests, the semen samples were kept at 37 degrees Celsius.
  • The aspect being closely examined was the sperm DNA fragmentation index (sDFI) – a measure of the degree to which the sperm DNA is fragmented or broken apart.

Findings of the Research

  • The study revealed that chilling or cryopreserving the semen does not significantly impact the initial sDFI after collection.
  • A significant increase in sDFI was observed within one hour of incubation at 37 degrees Celsius, with over 50% of sDFI seen after six hours. After 48 hours, almost all sperm showed DNA damage.
  • The rate of change of sDFI varied between individual stallions but not due to the preservation method.
  • The most intense period of DNA damage took set during the first six hours of incubation.
  • This suggests that chill or freeze-thaw preservation methods contribute to the fragmentation of sperm DNA, affecting the success of artificial insemination programs.

Implications and Recommendations

  • Using the SCD test can significantly help researchers understand the fertility potential of individual stallions.
  • These results allow for improved development and refinement of preservation protocols designed to reduce DNA damage in stallion sperm, potentially increasing the success rate of artificial insemination programs.

Cite This Article

APA
López-Fernández C, Crespo F, Arroyo F, Fernández JL, Arana P, Johnston SD, Gosálvez J. (2007). Dynamics of sperm DNA fragmentation in domestic animals II. The stallion. Theriogenology, 68(9), 1240-1250. https://doi.org/10.1016/j.theriogenology.2007.08.029

Publication

Theriogenology
ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 68
Issue: 9
Pages: 1240-1250

Researcher Affiliations

López-Fernández, C
  • Departamento de Biología, Unidad de Genética, Universidad Autónoma de Madrid (UAM), 20849 Madrid, Spain.
Crespo, F
    Arroyo, F
      Fernández, J L
        Arana, P
          Johnston, S D
            Gosálvez, J

              MeSH Terms

              • Analysis of Variance
              • Animals
              • Cryopreservation / methods
              • Cryopreservation / veterinary
              • DNA Fragmentation
              • Genetic Techniques / veterinary
              • Horses / physiology
              • Male
              • Microscopy, Fluorescence / veterinary
              • Nucleotides / metabolism
              • Refrigeration / veterinary
              • Regression Analysis
              • Reproductive Techniques, Assisted / veterinary
              • Semen / physiology
              • Semen Preservation / methods
              • Semen Preservation / veterinary
              • Spermatozoa / physiology
              • Time Factors

              Citations

              This article has been cited 17 times.
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