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The cryoprotective effect of Ficoll on the rabbit spermatozoa quality

Published online by Cambridge University Press:  26 September 2014

Barbora Kuliková*
Affiliation:
Constantine the Philosopher University, Slovak Republic Research Institute for Animal Production, NAFC, Nitra, Slovak Republic
Michele Di Iorio
Affiliation:
University of Molise, Campobasso, Italy
Elena Kubovicova
Affiliation:
Research Institute for Animal Production, NAFC, Nitra, Slovak Republic
Lenka Kuzelova
Affiliation:
Research Institute for Animal Production, NAFC, Nitra, Slovak Republic Slovak University of Agriculture, Nitra, Slovak Republic
Nicolaia Iaffaldano
Affiliation:
University of Molise, Campobasso, Italy
Peter Chrenek
Affiliation:
Research Institute for Animal Production, NAFC, Nitra, Slovak Republic Slovak University of Agriculture, Nitra, Slovak Republic
*
All correspondence to: Barbora Kuliková. Constantine the Philosopher University, Slovak Republic. Tel: +421 37 6546 289; Fax: +421 37 6546 285; e-mail: [email protected]

Summary

The aim of this study was to evaluate the effect of the addition of Ficoll 70 into the cryopreservation medium containing sucrose and dimethyl sulfoxide (DMSO) on rabbit spermatozoa characteristics following freezing/thawing. This large molecular weight polymer elevates the viscosity of medium and, therefore, could better protect spermatozoa during the freezing process. Only ejaculates of good initial motility (>80%) were used in the experiments. Heterospermic pools were diluted in a freezing medium composed of commercial diluent, 16% dimethyl sulphoxide (DMSO) and 2% sucrose (control) or in the same medium enriched with 4% Ficoll 70 (Ficoll) and frozen in liquid nitrogen vapours for 10 min before being plunged in liquid nitrogen. The quality of fresh and frozen/thawed spermatozoa samples was evaluated in vitro using the Computer Assisted Semen Analysis (CASA) system, fluorescent probes (peanut agglutinin (PNA)-Alexa Fluor®; annexin V-FLOUS) and by electron microscopy. Better cryoprotective effect was observed when Ficoll 70 was added, compared with the semen cryopreserved with sucrose and DMSO only. The higher values (P < 0.05) of motile and progressively moving spermatozoa immediately after thawing and at 30 min following incubation at 37°C were obtained in the Ficoll group. Moreover, the higher number (P < 0.05) of acrosome intact sperm was found in the Ficoll compared with the control group. Furthermore, no significant differences in kindling rates and number of pups born between frozen/thawed and fresh semen group were found. In conclusion, our study showed that the addition of Ficoll 70 might improve several characteristics of rabbit spermatozoa measured in vitro following freezing/thawing.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2014 

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References

An, T.Z., Iwakiri, M., Edashige, K., Sakurai, T. & Kasai, M. (2000). Factors affecting survival of frozen-thawed mouse spermatozoa. Cryobiology 40, 237–49.CrossRefGoogle ScholarPubMed
Castellini, C, Battaglini, M. & Lattaioli, P. (1992). Effects of cryoprotectants and freezing on rabbit semen quality. J. Appl. Rabbit Res. 15, 431–38.Google Scholar
Chen, Y. & Foote, R.H. (1994). Survival of rabbit spermatozoa frozen and thawed at different rates with and without seeding. Anim. Reprod. Sci. 35, 131–43.CrossRefGoogle Scholar
Chen, Y., Li, J., Simkin, M.E., Yang, X. & Foote, R.H. (1989). Fertility of fresh and frozen rabbit semen inseminated at different times is indicative of male differences in capacitation time. Biol. Reprod. 41, 848–53.CrossRefGoogle ScholarPubMed
Cheng, F.P., Fazeli, A., Voorhout, W.F., Marks, A., Bevers, M.M. & Colenbrander, B. (1996). Use of peanut agglutinin to assess the acrosomal status and the zona pellucida-induced acrosome reaction in stallion spermatozoa. J. Androl. 17, 674–82.CrossRefGoogle ScholarPubMed
Chrenek, P., Makarevich, A.V. & Simon, M. (2010). Viability and apoptosis in spermatozoa of transgenic rabbits. Zygote 20, 33–7.CrossRefGoogle ScholarPubMed
Curry, M.R., Redding, B.J. & Watson, P.F. (1995). Determination of water permeability coefficient and its activation energy for rabbit spermatozoa. Cryobiology 32, 175–81.CrossRefGoogle ScholarPubMed
Dalimata, A.M. & Graham, J.K. (1997). Cryopreservation of rabbit spermatozoa using acetamide in combination with trehalose and methyl cellulose. Theriogenology 48, 831–41.CrossRefGoogle ScholarPubMed
Darin-Bennett, A. & White, I.G. (1997). Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock. Cryobiology. 14, 466–70.CrossRefGoogle Scholar
Del Olmo, E., Bisbal, A., Maroto-Morales, A., García-Alvarez, O., Ramon, M., Jimenez-Rabadan, P., Martínez-Pastor, F., Soler, A.J., Garde, J.J. & Fernandez-Santes, M.R. (2013). Fertility of cryopreserved ovine semen is determined by sperm velocity. Anim. Reprod. Sci. 138, 102–9.CrossRefGoogle ScholarPubMed
Dumoulin, J.C., Janssen, J.M.B., Pieters, M.H., Enginsu, M.E., Geraedt, J.P. & Evers, J.L. (1994). The protective effects of polymers in the cryopreservation of human and mouse zonae pellucidae and embryos. Fertil. Steril. 62, 793–98.CrossRefGoogle ScholarPubMed
Gadella, B.M. & Harrison, R.A.P. (2002). Capacitation induces cyclic adenosine 3–5-monophosphate - dependent, but apoptosis - unrelated, exposure of aminophospholipids at the apical head plasma membrane of boar sperm cells. Biol. Reprod. 6, 640–50.Google Scholar
Hagen, D.R., Gilkey, A.L. & Foote, R.H. (2003). Spermatozoal velocity and motility and its relationship to fertility in rabbit inseminated with low sperm numbers. World Rabbit Sci. 10, 135–40.Google Scholar
Hirano, Y., Shibahara, H., Obara, H., Suzuki, T., Takamizawa, S., Jamaguchi, C., Tsunoda, H. & Sato, I. (2001). Relationship between sperm motility characteristics assessed by the computer–aided sperm analysis (CASA) and fertilization rates in vitro. J. Assist. Reprod. Genet. 18, 213–18.CrossRefGoogle ScholarPubMed
Iaffaldano, N., Di Iorio, M. & Rosato, M.P. (2012). The cryoprotectant used, its concentration, and the equilibration time are critical for the successful cryopreservation of rabbit sperm: dimethylacetamide versus dimethylsulfoxide. Theriogenology 78, 1381–89.CrossRefGoogle ScholarPubMed
Kapucinski, J. (1995). DAPI: a DNA-specific fluorescence probe. Biotech. Histochem. 70, 220–33.CrossRefGoogle Scholar
Kasai, M., Hamaguchi, Y., Zhu, S.E., Miyake, T., Sakurai, T. & Machida, T. (1992). High survival of rabbit morulae after vitrification in an ethylene glycol-based solution by a simple method. Biol. Reprod. 46, 1042–46.CrossRefGoogle Scholar
Kuleshowa, L.L., Shaw, J.M. & Trounson, A.O. (2001). Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation. Cryobiology 43, 2131.CrossRefGoogle Scholar
Lavara, R., Mocé, E., Lavara, F., Viudes de Castro, M.P. & Vicente, J.S. (2005). Do parameters of seminal quality correlate with the results of on-farm inseminations in rabbits? Theriogenology 64, 1130–41.CrossRefGoogle ScholarPubMed
López-Gatius, F., Sances, G., Sancho, M., Yániz, J., Santolaria, P., Gutiérrez, R., Núñez, M., Núñez, J. & Soler, C. (2005). Effect of solid storage at 15°C on the subsequent motility and fertility of rabbit semen. Theriogenology 64, 252–60.CrossRefGoogle Scholar
Lukac, N., Bardos, L., Stawarz, R., Roychoudhury, S., Makarevich, A.V. & Chrenek, P. (2011). In vitro effect of nickel on bovine spermatozoa motility and annexin-V-labeled membrane changes. J. Appl. Toxicol. 31, 144–49.CrossRefGoogle ScholarPubMed
Makarevich, A.V., Chrenek, P., Olexikova, L., Popelkova, M., Turanova, Z., Ostro, A. & Pivko, J. (2008a). Post-thaw survival, cell death and actin cytoskeleton in gene-microinjected rabbit embryos after vitrification. Theriogenology 70, 675–81.CrossRefGoogle ScholarPubMed
Makarevich, A.V., Parkanyi, V., Ondruska, Ľ., Kubovicova, E., Flak, P., Slezakova, M., Pivko, J. & Rafay, J. (2008b). Evaluation of fertilizing capacity of rabbit sperm on the basis of annexin-V-labelled membrane changes. Slovak. J. Anim. Sci. 41, 15.Google Scholar
Makarevich, A.V., Spalekova, E., Olexikova, L., Lukac, N., Kubovicova, E. & Hegedusova, Z. (2011). Functional characteristics or ram cooling-stored spermatozoa under the influence of epidermal growth factor. Gen. Physiol. Biophys. 30, S36–43.CrossRefGoogle ScholarPubMed
Martin, G., Sabido, O., Durand, P. & Levy, R. (2004). Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol. Reprod. 71, 2837.CrossRefGoogle ScholarPubMed
Martin, S.J., Reutelingsperger, C.P., McGahon, A.J., Rader, J.A., van Schie, R.C., LaFace, D.M. & Green, D.R. (1995). Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J. Exp. Med. 182, 1545–56.CrossRefGoogle ScholarPubMed
Mocé, E. & Vicente, J.S. (2009). Rabbit sperm cryopreservation: a review. Anim. Reprod. Sci. 110, 124.CrossRefGoogle ScholarPubMed
Mocé, E., Lavara, R. & Vicente, J.S. (2003a). Effect of an asynchrony between ovulation and insemination on the results obtained after insemination with fresh or frozen sperm in rabbits. Anim. Reprod. Sci. 75, 107–18.CrossRefGoogle ScholarPubMed
Mocé, E., Vicente, J.S. & Lavara, R. (2003b). Effect of freezing–thawing protocols on the performance of semen from three rabbit lines after artificial insemination. Theriogenology 60, 115–23.CrossRefGoogle ScholarPubMed
Mocé, E., Lavara, R. & Vicente, J.S. (2010). Effect of cooling rate to 5°C, straw size and farm on fertilizing ability of cryopreserved rabbit sperm. Reprod. Domest. Anim. 45, e1–7.Google Scholar
Papis, K., Sypecka, J., Korwin-Kossakowski, M., Wenta-Muchalska, E. & Bilska, B. (2005). Banking of embryos of mutated, paralytic tremor rabbits by means of vitrification. Lab. Anim. 39, 284–89.CrossRefGoogle ScholarPubMed
Parrish, J.J. & Foote, R.H. (1986). Fertility of cooled and frozen rabbit sperm measured by competitive fertilization. Biol. Reprod. 35, 253–7.CrossRefGoogle ScholarPubMed
Peña, F.J., Johannisson, A., Wallgren, M. & Rodríguez-Martínez, H. (2003). Assessment of fresh and frozen/thawed boar semen using an annexin-V assay: a new method of evaluating sperm membrane integrity. Theriogenology 60, 677–89.CrossRefGoogle ScholarPubMed
Peña, F.J., Núñez-Martínez, J.M. & Morán, J.M. (2006). Semen technologies in dog breeding: an update. Reprod. Domest. Anim. 41, 21–9.CrossRefGoogle ScholarPubMed
Peña, A.I., Barrio, M., Becerra, J.J., Quintela, L.A. & Herradón, P.G. (2012). Motile sperm subpopulation in frozen-thawed dog semen: Changes after incubation in capacitating conditions and relationship with sperm survival after osmotic stress. Anim. Reprod. Sci. 133, 214–23.CrossRefGoogle ScholarPubMed
Pivko, J., Makarevich, A.V., Kubovicova, E., Riha, L., Sirotkin, A.V. & Matejasakova, E. (2009). Ultrastructural alternations in sperm heads under influence of several implementors to ram semen. Slovak J. Anim. Sci. 42, 149–54.Google Scholar
Polgár, Z.S., Virag, G.Y., Baranyai, B., Bodó, S.Z., Kovács, A. & Gócza, E. (2004). Evaluation. of effects of cryopreservation on rabbit spermatozoa membranes with trypan blue-giemsa staining. Proceedings of the 8th World Rabbit Congress; 7–10 September 2004; Puebla, Mexico.Google Scholar
Rigau, T., Farré, M., Balleste, J., Mogas, T., Peña, A. & Rodríguez-Gil, J. E. (2001). Effects of glucose and fructose on motility patterns of dog spermatozoa from fresh ejaculates. Theriogenology 56, 801–15.CrossRefGoogle ScholarPubMed
Roca, J., Martínez, S., Vázquez, J.M., Lucas, X., Parrilla, I. & Martínez, E.A. (2000). Viability and fertility of rabbit spermatozoa diluted in Tris-buffer extenders and stored at 15°C. Anim. Reprod. Sci. 64, 103–12.CrossRefGoogle Scholar
Rosato, M.P. & Iaffaldano, N. (2013). Cryopreservation of rabbit semen: comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification. Theriogenology 79, 508–16.CrossRefGoogle ScholarPubMed
Saffa, H.M., Vicente, J.S., Lavara, R. & Viudes de Castro, M.P. (2008). Semen evaluation of two selected lines of rabbit bucks. World Rabbit Sci. 16, 141–48.Google Scholar
Shaw, J.M., Kuleshowa, L.L., MacFarlane, D.R. & Trounson, A.O. (1997). Vitrification properties of solutions of ethylene glycol in saline containing PVP, Ficoll, or dextran. Cryobiology 35, 219–29.CrossRefGoogle ScholarPubMed
Siqueira, A.P., Wallgren, M., Hossain, M.S., Johannisson, A., Sanz, L., Calvete, J.J & Rodríguez-Martínez, H. (2011). Quality of boar spermatozoa from the sperm-peak portion of the ejaculate after simplified freezing in Mini Flatpacks compared to the remaining spermatozoa of the sperm-rich fraction. Theriogenology 75, 1175–84.CrossRefGoogle Scholar
Suarez, S.S., Katz, D.F., Owen, D.H., Andrew, J.B. & Powel, R.L. (1991). Evidence for the function of hyperactivated motility in sperm. Biol. Reprod. 44, 375–81.CrossRefGoogle ScholarPubMed
Swain, J.E. & Smith, G.D. (2010). Cryoprotectants. In Fertility Cryopreservation (eds Chian, R.C., & Quinn, P.), pp. 2438. UK: Cambridge University Press.CrossRefGoogle Scholar
Valcárcel, A., De las Heras, M.A., Pérez, L., Moses, D.F. & Baldassarre, H. (1997). Assessment of the acrosomal status of membrane-intact ram-spermatozoa after freezing and thawing, by simultaneous lectin/Hoechst 33258 staining. Anim. Reprod. Sci. 45, 299309.CrossRefGoogle ScholarPubMed
Vicente, J.S. & Viudes de Castro, M.P. (1996). A sucrose-DMSO extender for freezing rabbit semen. Reprod. Nutr. Dev. 36, 485–92.CrossRefGoogle ScholarPubMed