Stallion Sperm Selection by Density Gradient CentrifugationInvolving a Double Layer Colloid: Effects on Sperm Subpopulation Dynamics in Fresh and Stored Semen

Document Type: Research Article

Authors

1 Departmentof Animal Medicine and Surgery, Faculty of Veterinary, University of Autónoma de Barcelona, 08193 Bellaterra, Spain

2 Department of Celular Biology, Phisiology and Immunology, Faculty of Veterinary, University of Córdoba, 14071 Córdoba, Spain

Abstract

Sperm selection techniques have became an important tool to improve sub fertile ejaculates. The main aim of this study was to assess the dynamics of motile sperm subpopulations of ejaculates from subfertile stallions subjected to density gradient centrifugation (DGC) involving a double layer colloid [DGC2], before and after storage for 24 and 48 hours. Ejaculates from eight stallions with fertility problems were subjected to the different treatments: 1) control [C]; 2) centrifugation [CT]; 3) density gradient centrifugation involving a double layer colloid [DGC2] and 4) post storage density gradient centrifugation involving a double layer colloid DGC2 post]. In the C, CT and DGC2 treatments, sperm variables were analysed at 0 h and after 24 and 48 h of storage at 4-7 ˚C. In the DGC2 post treatment, the semen was stored at 4-7˚Cand sperm samples selected by DGC2 at 24 and 48 h. Viability, sperm abnormalities and motility were then examined using a computerized system. Four motile sperm subpopulations with different motility characteristics were observed in the all treatments. The DGC2 sperm had faster, straighter moving and more active cells at 0 h. At 24 and 48 h, however, the motility variable values fell and the slower spermatozoa subpopulations increased in size. The DGC2 post treatment returned better overall sperm motility variable values at 24 and 48 h; a larger percentage of spermatozoa fell into the faster subpopulations. In conclusion, DGC2 could be used to improve semen from subfertile stallions as well as semen showing poor fertility after storage.

Keywords

References

Abaigar T., Holt W.V., Harrison R.A. and Del Barrio G. (1999). Sperm subpopulations in boar (Sus scrofa) and gazelle (Gazella dama mhorr) semen as revealed by pattern analysis of computer-assisted motility assessments. Biol. Reprod. 60, 32-41.
Abaigar T., Cano M., Pickard A.R. and Holt W.V. (2001). Use of computerized-assisted sperm motility assessment and multivariate pattern analysis to characterize ejaculate quality in mohor gazelles (Gazella dama mhorr): effects of body weight, electro ejaculation technique and short-term semen storage. Reproduction. 122, 265-273.
Crosier A.E., Henghali J.N., Howard J., Pukazhenthi B.S., Terrell K.A., Marker L.L. and Wildt D.E. (2008). Improved quality of cryopreserved cheetah (Acinonyx jubatus) spermatozoa after centrifugation through accudenz.J. Androl. 30, 298-308.
Fleming S.D., Ilad R.S., Griffin A.M., Wu Y., Ong K.J., Smith H.C. and Aitken R.J. (2008). Prospective controlled trial of an electrophoretic method of sperm preparation for assisted reproduction: comparison with density gradient centrifugation. Hum. Reprod. 23, 2646-2651.
Flores E., Taberner E., Rivera M.M., Pena A., Rigau T., Miro J. and Rodriguez-Gil J.E. (2008). Effects of freezing / thawing on motile sperm subpopulations of boar and donkey ejaculates. Theriogenology. 70, 936-945.
Hammoud A.O., Gibson M., Peterson M.C. and Carrell D.T. (2007). Effect of sperm preparation techniques by density gradient on intra-individual variation of sperm motility. Arch. Androl. 53, 349-351.
Johannisson A., Morrell J.M., Thoren J., Jonsson M., Dalin A.M. and Rodriguez-Martinez H. (2009). Colloidal centrifugation with androcoll-E prolongs stallion sperm motility, viability and chromatin integrity. Anim. Reprod. Sci. 116, 119-128.
Keeney R.M., Bergman R.V., Cooper W.L. and Morse G.W. (1975). Minimal contamination techniques for breeding mares: Technique and preliminary finding. Pp. 327-336 in 22th Ann. Conv. Proc. Am. Assoc. Eq. Pract.
Lopez-Fernandez C., Crespo F., Arroyo F., Fernández J.L., Arana P., Johnston S.D. and Gosálvez J. (2007). Dynamics of sperm DNA fragmentation in domestic animals: II. The stallion. Theriogenology. 68, 1240-1250.
Macias Garcia B., Morrell J.M., Ortega-Ferrusola C., Gonzalez-Fernandez L., Tapia J.A., Rodriguez-Martinez H. and Pena F.J. (2009). Centrifugation on a single layer of colloid selects improved quality spermatozoa from frozen-thawed stallion semen. Anim. Reprod. Sci. 114, 193-202.
Macias Garcia B., Gonzalez Fernandez L., Morrell J.M., Ortega Ferrusola C., Tapia J.A., Rodriguez Martinez H. and Pena F.J. (2005). Single-layer centrifugation through colloid positively modifies the sperm subpopulation structure of frozen-thawed stallion spermatozoa. Reprod. Domest. Anim. 44, 523-526.
Martinez-Pastor F., Garcia-Macias V., Alvarez M., Herraez P., Anel L. and de Paz P. (2005). Sperm subpopulations in Iberian red deer epididymal sperm and their changes through the cryopreservation process. Biol. Reprod. 72, 316-327.
Miró J., Lobo V., Quintero-Moreno A., Medrano A., Peña A. and Rigau T. (2009). Sperm motility patterns and metabolism in catalonian donkey semen. Theriogenology. 63, 1706-1716.
Miró J., Taberner E., Rivera M., Peña A., Medrano A., Rigau T. and Peñalba A. (2005). Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen. Theriogenology. 72, 1017-1022.
Morató R., De Souza J., Orero G., Mogas T. and Miró J. (2013). Pre-selection double layer density gradient centrifugation improves the fertilising capacity of frozen-thawed, capacitated stallion sperm. Anim. Reprod. Sci. 139, 62-68.
Morrell J.M., Saravia F., VAN Wienen M., Wallgren M. and Rodriguez-Martinez H. (2009a). Selection of boar spermatozoa using centrifugation on a glycidoxypropyltrimethoxysilane-coated silica colloid. J. Reprod. Dev. 55, 547-552.
Morrell J.M., Johannisson A., Dalin A.M. and Rodriguez-Martinez H. (2009b). Morphology and chromatin integrity of stallion spermatozoa prepared by density gradient and single layer centrifugation through silica colloids. Reprod. Domest. Anim. 44, 512-517.
Morrell J.M., Johannisson A., Strutz H., Dalin A. and Rodriguez-Martinez H. (2009c). Colloidal centrifugation of stallion semen: changes in sperm motility, velocity and chromatin integrity during storage. J. Equine Vet. Sci. 29, 24-32.
Morrell J.M., Dalin A.M. and Rodriguez-Martinez H. (2009d). Comparison of density gradient and single layer centrifugation of stallion spermatozoa: yield, motility and survival. Equine Vet. J. 41, 53-58.
Morrell J.M., Garcia B.M., Pena F.J. and Johannisson A. (2011). Processing stored stallion semen doses by single layer centrifugation. Theriogenology. 76, 1424-1432.
Quintero-Moreno A., Miró J., Teresa Rigau A. and Rodrı́guez-Gil J.E. (2003). Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology. 59, 1973-1990.
SAS Institute. (1996). SAS®/STAT Software, Release 6.11. SAS Institute, Inc., Cary, NC. USA.
Sieme H., Martinsson G., Rauterberg H., Walter K., Aurich C., Petzoldt R. and Klug E. (2003). Application of techniques for sperm selection in fresh and frozen-thawed stallion semen. Reprod. Domest. Anim. 38, 134-140.
De SouzaSoares J., Morató R., Orero G., Mogas T. and Miró J. (2001). Assessment of different capacitation treatments to evaluate the ability of non-filtered and filtered frozen-thawed stallion sperm to penetrate zona pellucida-free bovine oocytes matured in vitro. Pp.134-136 in Proc. 27th Scientific Meeting AETE, Chester, England.
Thys M., Vandaele L., Morrell J.M., Mestach J., Van Soom A., Hoogewijs M. and Rodriguez-Martinez H. (2008). In vitro fertilizing capacity of frozen-thawed bull spermatozoa selected by single-layer (glycidoxypropyltrimethoxysilane) silane-coated silica colloidal centrifugation. Reprod. Domest. Anim. 44, 390-394.
Varner D.D., Love C.C., Brinsko S.P., Blanchard T.L., Hartman D.L., Bliss S.B., Carroll B.S. and Eslick M.C. (2008). Semen processing for the subfertile stallion. J. Equine Vet. Sci. 28, 677-685.
 

Iranian Journal of Applied Animal Science

Volume 5, Issue 2
June 2015
Pages 317-321

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History

  • Receive Date: 27 July 2014
  • Revise Date: 08 September 2014
  • Accept Date: 15 November 2014

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