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2014 | 70 | 09 |

Tytuł artykułu

Quality of cooled semen of cold-blooded stallions evaluated with the use of apoptosis and DNA defragmentation markers

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
The aim of this study was to determine the quality of semen collected from stallions and chilled to +4°C. Semen was collected from eight Polish cold-blooded stallions, aged 4-8 years. The ejaculate was examined macro- and microscopically to evaluate volume, concentration, viable sperm percentage, progressive motility, and morphology. The percentage of live/dead spermatozoa was determined by eosin and nigrosin staining. Flow cytometry was applied to detect phosphatidylserine translocation, whereas DNA defragmentation was determined with a commercial kit. The semen samples were examined immediately after collection and dilution (time 0) and at 6 storage times: 24, 48, 72, 96, and 120 hours later. Time-related changes in all the investigated spermatozoa parameters were observed at 6 different times. Progressive motility was negatively correlated with the percentage of live spermatozoa with phosphatidylserine translocation, but positively correlated with DNA defragmentation (P < 0.05). This study demonstrates that the evaluation of selected indicators of apoptosis is very useful in assessing the quality of chilled stallion semen. The results obtained show what significant changes occur in spermatozoa during storage at +4°C. Evaluation of the cell membrane integrity and DNA defragmentation of spermatozoa makes it possible to determine whether and for how long semen chilled to +4°C may be used for artificial insemination, which is extremely important during semen transportation. The methods of semen quality evaluation applied in this study should also be used in determining the usefulness of chilled semen for artificial insemination. This may lead to higher fertilization rates in mares.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

70

Numer

09

Opis fizyczny

p.572-577,ref.

Twórcy

autor
  • Department of Andrology and Reproductive Biotechnology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, ul. Gleboka 30, 20-612 Lublin, Poland
autor
  • Department of Andrology and Reproductive Biotechnology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, ul. Gleboka 30, 20-612 Lublin, Poland
autor
  • Department of Andrology and Reproductive Biotechnology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, ul. Gleboka 30, 20-612 Lublin, Poland
  • Department of Cell Biology, Marie Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland
autor
  • Department of Animal Physiology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, ul. Akademicka 12, 20-950 Lublin, Poland
autor
  • Department of Andrology and Reproductive Biotechnology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, ul. Gleboka 30, 20-612 Lublin, Poland

Bibliografia

  • 1. Anzar M., He L., Buhr M. M., Kroetsch T. G., Pauls K. P.: Sperm apoptosis in fresh and cryopreserved bull semen detected by flow cytometry and its relationship with fertility. Biol. Reprod. 2002, 66, 354-360.
  • 2. Ball B. A.: Oxidative stress, osmotic stress and apoptosis: Impacts on sperm function and preservation in the horse. Anim. Reprod. Sci. 2008, 107, 257-267.
  • 3. Blanco-Rodrigues J., Martinez-Gracia C., Porras A.: Correlation between DNA synthesis in the second, third and fourth generation of spermatogonia and the occurrence of apoptosis in both spermatogonia and spermatocytes. Reproduction 2003, 126, 661-668.
  • 4. Brum A. M., Sabeur K., Ball B. A.: Apoptotic-like changes in equine spermatozoa separated by density-gradient centrifugation or after cryopreservation. Theriogenology 2008, 69, 1041-1055.
  • 5. Evenson D., Jost L.: Sperm chromatin structure assay is useful for fertility assessment. Methods Cell Sci. 2000, 22, 169-189.
  • 6. Evenson D. P., Wixon R.: Predictive value of the sperm chromatin assay in different population. Fertil. Steril. 2006, 85, 811-812.
  • 7. Foster M. L., Varner D. D., Hinrichs K., Teague S., LaCaze K., Blanchard T. L., Love C. C.: Agreement between measures of total motility and membrane integrity in stallion sperm. Theriogenology 2011, 75, 1499-1505.
  • 8. Freitas-Dell’Aqua C. P., Monteiro G. A., Dell’Aqua J. A. Jr, Papa F. O.: The effects of refrigeration temperature and storage time on apoptotic markers in equine semen. J. Equine Vet. Sci. 2012, 30, 1-5.
  • 9. Gallardo-Bolanos J. M., Moran A. M., Balao de Silva C. M., Rodriguez A. M., Davila M. P., Aparicio I. N., Tapia J. A., Ortega-Ferrusola C., Pena F. J.: Autophagy and apoptosis have a role in the survival or death of stallion spermatozoa during conservation in refrigeration. PLoS ONE 2012, 7, e30688.
  • 10. Giannoccaro A., Lacalandra G. M., Filannino A., Pizzi F., Nicassio M., Dell’Aquila M. E., Minervini F.: Assessment of viability, chromatin structure stability, mitochondrial function and motility of stallion fresh sperm by using objective methodologies. J. C. A. B. 2010, 4, 034-041.
  • 11. Gorczyca W.: Cytometric analyses to distinguish death processes. Endocr. Relat. Cancer 1999, 6, 17-19.
  • 12. Heckenbichler S., Deichsel K., Peters P., Aurich C. H.: Quality and fertility of cooled-shipped stallion semen at the time of insemination. Theriogenology 2011, 75, 849-856.
  • 13. Janett F., Thun R., Bettschen S., Burger D., Hassig M.: Seasonal changes of semen quality and freezability in Franches-Montagnes stallion. Anim. Reprod. Sci. 2003, 77, 213-221.
  • 14. Krakowski L., Obara J., Wąchocka A., Piech T., Bartoszek P., Kostro K., Tatara M. R.: Assessment of extent of apoptosis and DNA defragmentation in chilled semen of stallions during the breeding season. Reprod. Domest. Anim. 2013, 48, 826-832.
  • 15. Lachaud C. H., Tesarik J., Canadas M. L., Mendoza C.: Apoptosis and necrosis in human ejaculated spermatozoa. Hum. Reprod. 2004, 19, 607-610.
  • 16. Love C. C.: The sperm chromatin structure assay: A review of clinical applications. Anim. Reprod. Sci. 2005, 89, 39-45.
  • 17. Love C. C., Thompson J. A., Lowry V. K., Varner D. D.: Effect of storage time and temperature on stallion sperm DNA and fertility. Theriogenology 2002, 57, 1135-1142.
  • 18. Love C. C., Thompson J. A., Lowry V. K., Varner D. D.: The relationship between chromatin quality and fertility of chilled stallion sperm. A. A. E. P. 2001, 47, 229-231.
  • 19. López-Fernández C., Crespo F., Arroyo F., Fernández J. L., Arana P., Johnston S. D., Gosálvez J.: Dynamics of sperm DNA fragmentation in domestic animals II. The Stallion. Theriogenology 2007, 68, 1240-1250.
  • 20. Merkies K., Chenier T., Plante C., Buhr M. M.: Assessment of stallion spermatozoa viability by flow cytometry and light microscope analysis. Theriogenology 2000, 54, 1215-1224.
  • 21. Morrel J. M., Jahannisson A., Dalin A. M., Hammar L., Sandebert T., Rodriguez-Martinez H.: Sperm morphology and chromatin integrity in Swedish warmblood stallion and their relationship to pregnancy rates. Acta Vet. Scand. 2008, 50, 2.
  • 22. Oosterhuis G. J. E., Verme I.: Apoptosis in human ejaculated spermatozoa. J. Biol. Regul. Homeost. Agents 2004, 18, 115-119.
  • 23. Ortega-Ferrusola C., Macias Garcia B., Gallardo-Bolanos J. M., González-Fernández L., Rodriguez-Marinez H., Tapia J. A., Pena F. J.: Apoptotic markers can be used to forecast freezeability of stallion spermatozoa. Anim. Reprod. Sci. 2009, 114, 393-403.
  • 24. Ortega-Ferrusola C., Sotillo-Galán Y., Varela-Fernández E., Gallardo-Bolanos J. M., Muriel A., González-Fernández L., Tapia J. A., Pena F. J.: Detection of “apoptosis-like” changes during the cryopreservation process in equine sperm. J. Androl. 2007, 29, 213-221.
  • 25. Rodriguez-Martinez H.: State of the art in farm animal sperm evaluation. Reprod. Fertil. Dev. 2007, 19, 91-101.
  • 26. Varner D. D.: Developments in stallion semen evaluation. Theriogenology 2008, 70, 448-462.
  • 27. Varner D. D., Blanchard T. L., Love C. L., Garci M. C., Kenney R. M.: Effects of cooling rate and storage temperature on equine spermatozoal motility parameters. Theriogenology 1988, 29, 1043-1054.

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Typ dokumentu

Bibliografia

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