Influence of glycerol addition on the quality of cat epididymal sperm during the freeze-thaw process
DOI:
https://doi.org/10.5433/1679-0359.2020v41n4p1237Keywords:
Cryo-injury, Cryopreservation, Sperm, Feline, Intracellular cryoprotectant.Abstract
Cryopreservation of epididymal sperm is a useful tool for preserving the genetic potential of valuable animal specimens. The domestic cat is used as a model to study and develop cryogenics for other felines. However, regulation of the entire cryopreservation process is essential for the success of this biotechnology. Thus, our aim was to evaluate the effects of glycerol equilibration time and freeze-thaw stages on the quality of epididymal sperm obtained from domestic cats. Epididymal sperm were recovered with TRIS and immediately evaluated for total motility (TM), vigor, viability, membrane functionality (HOST), and morphology. Then, TRIS-20% egg yolk was added to the samples, which were equally divided into two 1.5 mL tubes and refrigerated at 4 ºC for 1 hour. Subsequently, glycerol was added at a final concentration of 5%. The samples were incubated with glycerol (equilibration time) for either 5 or 10 minutes (groups G5 and G10, respectively) and then frozen. Thawing occurred at 37 ºC for 30 seconds. The samples were evaluated at all stages. A reduction in TM was observed only after thawing; however, it was higher in G5 (39.00 ± 4.07%) than in G10 (18.50 ± 4.54%). Vigor declined in both groups after thawing; however, they did not differ from each other. Sperm viability was maintained in G5 after glycerolization (53.60 ± 2.59%); in G10, sperm viability decreased in the glycerolized sample (48.80 ± 2.93%) when compared to that in the fresh sample (59.90 ± 1.74%). Post-thaw viability of G5 (33.80 ± 1.89%) was higher than that of G10 (18.80 ± 3.01%). In the HOST, a decrease in viability was only observed after thawing, with no difference between the groups (41.50 ± 2.84% for G5 and 40.20 ± 3.49% for G10). With regard to sperm morphology, normal sperm decreased while sperm with post-thaw secondary defects increased in both groups. In conclusion, a shorter equilibration time for glycerolization preserves epididymal sperm quality better and the freeze-thaw process is the most critical stage of thawing.Downloads
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References
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Axnér, E., Hermansson, U., & Linde-Forsberg, C. (2004). The effect of Equex STM paste and sperm morphology on post-thaw survival of cat epididymal spermatozoa. Animal Reproduction Science, 84(1-2), 179-191. doi:10.1016/j.anireprosci.2003.11.003
Bogliolo, L., Bonelli, P., Madau, L., Zedda, M. T., Santucciu, C., Leoni, G.,... Ledda, S. (2004). Fertilizing ability of epididymal cat spermatozoa after cryopreservation or storage at 4 °C. Reproduction, Fertility and Development, 16(2), 222-222. doi: 10.1071/RDv16n1Ab202
Buranaamnuay, K. (2015). Determination of appropriate cryopreservation protocols for epididymal cat spermatozoa. Reproduction in Domestic Animals, 50(3), 378-385. doi: 10.1111/rda.12496
Buranaamnuay, K. (2017). Protocols for sperm cryopreservation in the domestic cat: a review. Animal Reproduction Science, 183(1), 56-65. doi: 10.1016/j.anireprosci.2017.06.002
Buranaamnuay, K. (2018). Cryopreservation and storage of cat epididymal sperm using‒75° C freezer vs liquid nitrogen. Animal Reproduction Science, 191(1), 56-63. doi: 10.1016/j.anireprosci.2018.02.008
Burg, M. B., Ferraris J. D., & Dmitrieva, N. I. (2007). Cellular response to hyperosmotic stresses. Physiological Reviews, 87(4), 1441-1474. doi: 10.1152/physrev.00056.2006
Colégio Brasileiro de Reprodução Animal (2013). Manual para exame andrológico e avaliação de sêmen animal. 3a ed. Belo Horizonte, MG: CBRA.
Chatdarong, K. (2017). Retained fertilizing capability in cryopreserved feline spermatozoa. Reproduction in Domestic Animals, 52(2), 261-264. doi: 10.1111/rda.12855
Cheuquemán, C., Faúndez, R., Sánchez, R., & Risopatrón, J. (2018). Changes in sperm function and structure after freezing in domestic cat spermatozoa. Andrologia, 50(9), e13080. doi: 10.1111/and.13080
Cheuquemán, C., Sánchez, R., & Risopatrón, J. (2017). Effect of sperm selection techniques in frozen/thawed cat spermatozoa on sperm motility analyzed by CASA System. International Journal of Morphology, 35(4), 1495-1501. doi: 10.4067/S0717-95022017000401495
Cocchia, N, Ciani, F., El-Rass, R., Russo, M., Borzacchiello, G., Esposito, V.,... Lorizio, R. (2010). Cryopreservation of feline epididymal spermatozoa from dead and alive animals and its use in assisted reproduction. Zygote, 18(1), 1-8. doi: 10.1017/S0967199409990256
Costa, L. L. M., Castelo, T. S., Souza, A. L. P., Lima, G. L., & Silva, A. R. (2013). Criopreservação de sêmen canino em diluente Tris adcionado de dodecil sulfato de sódio. Revista Brasileira Reprodução Animal, 37(1), 53-58. Recuperado de https://cbra.websiteseguro.com/pages/publicacoes/rbra/v37n1/p53-58%20 (RB386).pdf
Deco-Souza, T., Paula, T. A. R., Costa, D. S., Costa, E. P., Barros, J. B. G., Araujo, G. R., & Carreta, M., Jr. (2013). Comparação entre duas concentrações de glicerol para a criopreservação de sêmen de suçuarana (Puma concolor). Pesquisa Veterinária Brasileira, 33(4), 512-516. doi: 10.1590/S0100-736X2013000400015
Di Domenico, D., Pedroso, E. M. S. R., & Teixeira, P. P. M. (2015) Diferenças da célula espermática suína e crioprotetores: Revisão. Nucleus Animalium, 7(1), 1-5. doi: 10.3738/1982.2278.1169
Figueroa, E., Valdebenito, I., & Farias, J. G. (2016). Technologies used in the study of sperm function in cryopreserved fish spermatozoa. Aquaculture Research, 47(6), 1691-1705. doi: 10.1111/are.12630
García, B. M., Ferrusola, C. O., Aparicio, I. M., Miró-Morán, A., Morillo-Rodriguez, A., Bolaños, J. M. G.,… Peña, F. J. (2012). Toxicity of glycerol for the stallion spermatozoa: effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential. Theriogenology, 77(7), 1280-1289. doi: 10.1016/j.theriogenology.2011.10.033
García-Vazquez, F. A., Gadea, J., Matás, C., & Holt, W. V. (2016). Importance of sperm morphology during sperm transport and fertilization in mammals. Asian Journal of Andrology, 18(6), 844-850. doi: 10.4103/1008-682X.186880
Guerra, M. M. P., Souza, A. F., Soares, A. T., César, C. N. R., Silva, S. V., & Batista, A. M. (2009). Aspectos críticos da congelação do sêmen caprino. Revista Brasileira de Reprodução Animal, 1(Supl. 6), 42-49. Recuperado de https://cbra.websiteseguro.com/pages/publicacoes/rbra/download/p42-49.pdf
Hartwig, F. O. P., Papa, F. O., & Dell’Aqua, J. A. J. (2012). Utilização do colesterol na criopreservação de espermatozóides na espécie equina: uma revisão. Veterinária e Zootecnia, 19(2), 157-168. Recuperado de https://repositorio.unesp.br/bitstream/handle/11449/141277/ISSN0102-5716-2012-19-02-157-168.pdf?sequence=1&isAllowed=y
Jeyendran, R. S., Van Der Ven, H. H., Perez-Pelaez, M., Crabo, B. G., & Zaneveld, L. J. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Reproduction, 70(1), 219-28. doi: 10.1530/jrf.0.0700219
Jiménez, E., Pérez-Marín, C. C., Vizuete, G., Millán, Y., & Agüera, E. I. (2013). Effect of different extenders on in vitro characteristics of feline epididymal sperm during cryopreservation. Reproduction in Domestic Animals, 48(4), 665-672. doi: 10.1111/rda.12142
Klaus, C., Eder, S., Franz, C., & Müller, K. (2016). Successful cryopreservation of domestic cat (Felis catus) epididymal sperm after slow equilibration to 15 or 10°C. Reproduction in Domestic Animals, 51(2), 195-203. doi: 10.1111/rda.12666
Kunkitti, P., Chatdarong, K., Suwimonteerabutr, J., Nedumpun, T., Johannisson, A., Bergqvist, A.,… Axnér, E. (2017). Osmotic tolerance of feline epididymal spermatozoa. Animal Reproduction Science, 185(1), 148-153. doi: 10.1016/j.anireprosci.2017.08.014
Leite, T. G., Vale, V. R., Fº., Arruda, R. P., Andrade, A. F. C., Emerick, L. L., Zaffalon, F. G.,… Andrade, V. J. (2010). Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry. Animal Reproduction Science, 120(4), 31-38. doi.org/10.1016/j.anireprosci.2010.04.005
Luvoni, G. C. & Morselli, M. G. (2017). Canine epididymal spermatozoa: A hidden treasure with great potential. Reproduction in Domestic Animals, 52(2), 197-201. doi: 10.1111/rda.12820
Martins, J. L. A., Villaverde, A. I. S. B., Lima, A. F. M., Steagall, P. V. M., Ferreira, J. C. P., Taconeli, C. A., & Lopes, M. D. (2009). Impact of 24‐h cooling prior to freezing on the survival of domestic cat (Felis catus) epididymal sperm. Reproduction in Domestic Animals, 44(2), 366-368. doi: 10.1111/j.1439-0531.2009.01432.x
Oliveira, R. A. (2013). Criopreservação de sêmen equino, um desafio a ser vencido. PUBVET, 7(26), 2678-2755. Recuperado de http://www.pubvet.com.br
Oliveira, R. V., Nunes, J. F., Salgueiro, C. C. M., Cavalcante, J. M. M., Brasil, O. O., & Moura, A. A. A. N. (2009). Avaliação de espermatozóides caprinos congelados em meio à base de água de coco em pó (ACP-101®) ou TRIS. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 63(6), 1295-1302. doi: 10.1590/S010209352011000600003
Ozkavukcu, S., Erdemli, E., Isik, A., Oztuna, D., & Karahuseyinoglu, S. (2008). Effects of cryopreservation on sperm parameters and ultrastructural morphology of human spermatozoa. Journal of Assisted Reproduction and Genetics, 25(8), 403-411. doi: 10.1007/s10815-008-9232-3
Prochowska, S., Niżański, W., & Partyka, A. (2016). Comparative analysis of in vitro characteristics of fresh and frozen-thawed urethral and epididymal spermatozoa from cats (Felis domesticus). Theriogenology, 86(8), 2063-2072. doi: 10.1016/j.theriogenology.2016.07.002
Raskin, R. E., Meyer, D. J. (2011). Citologia Clínica de Cães e Gatos: Atlas Colorido e Guia de Interpretação (2a ed.). Rio de Janeiro: Elsevier Brasil.
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2020-05-13
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Barbosa, B. de S., Silva, H. V. R., Silva, T. F. P. da, & Silva, L. D. M. da. (2020). Influence of glycerol addition on the quality of cat epididymal sperm during the freeze-thaw process. Semina: Ciências Agrárias, 41(4), 1237–1246. https://doi.org/10.5433/1679-0359.2020v41n4p1237
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