Effects of the addition of docosahexaenoic acid and ?-tocopherol on quality of equine spermatozoa stored at 5°C

Breno Fernandes Barreto Sampaio, Bruno Gomes Nogueira, Maria Inês Lenz Souza, Eliane Vianna da Costa-e-Silva, Carmem Estefânia Serra Neto Zúccari

Abstract


Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.

Keywords


Antioxidant; DHA; Stallion; Vitamin E.

Full Text:

PDF

References


Agarwal, A., & Said, T. M. (2003). Role of sperm chromatin abnormalities and DNA damage in male infertility. Human Reproduction, 9(4), 331-345. doi: 10.1093/humupd/dmg027

Agarwal, A., Saleh, R. A., & Bedaiwy, M. A. (2003). Role of reactive oxygen species in the pathophysiology of human reproduction. Fertilily and Sterility, 79(4), 829-843. doi: 10.1016/S0015-0282(02) 04948-8

Amann, R. P., & Pickett, B. W. (1987). Principles of cryopreservation and a review of cryopreservation of stallion spermatozoa. Journal of Equine Veterinary Science, 7(3), 145-173. doi: 10.1016/S0737-0806(87)80025-4

Almeida, J., & Ball, B. A. (2005). Effect of alfa tocopherol and tocopherol succinate on lipid peroxidation in equine spermatozoa. Animal Reproduction Science, 87(3-4), 321-337. doi: 10.1016/j.anireprosci.2004.12.004

Aurich, C. (2005). Factors affecting the plasma membrane function of cooled-stored stallion spermatozoa. Animal Reproduction Science, 89(1-4), 65-75. doi: 10.1016/j.anireprosci.2005.06.025

Aurich, C. (2008). Recent advances in cooled-semen technology. Animal Reproduction Science, 107(3-4), 268-275. doi: 10.1016/j.anireprosci.2008.04.015

Ball, B. A., Medina, V., Gravance, C. G., & Baumber, J. (2001). Effect of antioxidants on preservation of motility, viability and acrosomal integrity of equine spermatozoa during storage at 5°C. Theriogenology, 56(4), 577-589. doi: 10.1016/S0093-691X(01)00590-8

Ball, B. A., & Vo, A. (2002). Detection of lipid peroxidation in equine spermatozoa based upon the lipophilic fluorescent dye C-11-BODIPY581/591. Journal of Andrology, 23(2), 259-269. doi: 10.1002/j.1939-4640.2002.tb02623.x

Ball, B. A. (2008). Oxidative stress, osmotic stress and apoptosis: impacts on sperm function and preservation in the horse. Animal Reproduction Science, 107(3-4), 257-267. doi: 10.1016/j.anireprosci.2008.04.014

Baumber, J., Ball, B. A., & Linfar, J. J. (2005). Assessment of the cryopreservation of equine spermatozoa in the presence of enzyme scavengers and antioxidants. American Journal of Veterinary Research, 66(5), 772-779. doi: 10.2460/ajvr.2005.66.772

Beletti, M. E., Costa, L. F., & Viana, M. P. (2004). Computational approach to characterization of bovine sperm chromatin alterations. Biotechnic and Histochemistry, 79(1), 17-23. doi: 10.1080/10520290410001700774

Brinsko, S. P., Varner, D. D., Love, C. C., Blanchard, T. L., Day, B. C., & Wilson, M. E. (2005). Effect of feeding DHA enriched nutraceutical on the quality of fresh, cooled and frozen stallion semen. Teriogenology, 63(5), 1519-1527. doi: 10.1016/j.theriogenology.2004.07.010

Castellini, C., Lattaioli, P., Moroni, M., & Minelli, A. (2000). Effect of seminal plasma on the characteristics and fertility of rabbit spermatozoa. Animal Reproduction Science, 63(3-4), 275-282. doi: 10.1016/S0378-4320(00)00181-0

Cerolini, S., Maldjian, A., Surai, P., & Noble, R. (2000). Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage. Animal Reproduction Science, 58(1-2), 99-111. doi: 10.1016/S0378-4320(99)00035-4

Connor, W. E., Lin, D. S., Wolf, D. P., & Alexander, M. (1998). Uneven distribution of desmosterol and docosahexaenoic acid in the heads and tails of monkey sperm. Journal of Lipid Research, 39(7), 1404-1411.

Dodaran, H. V., Zhandi, M., Sharafi, M., Nejati-Amiri, E., Nejati-Javaremi, A., Mohammadi-Sangcheshmeh, A., Shakeri, M. (2015). Effect of ethanol induced mild stress on post thawed bull sperm quality. Cryobiology, 71(1), 12-17. doi: 10.1016/j.cryobiol.2015.06.008

Evenson, D. P. & Jost, L. (2000). Sperm chromatin structure assay is useful for fertility assessment. Methods in Cell Science, 22(2-3), 169-189.

Franco, J. S. V., Chaveiro, A., Góis, A., & Silva, F. M. (2013). Effects of alfa tocopherol and ascorbic acid on equine semen quality after cryopreservation. Journal of Equine Veterinary Science, 33(10), 787-793. doi: 10.1016/j.jevs.2012.12.012

Gawrisch, K., Eldho, N. V., & Holte, L. L. (2003). The structure of DHA in phospholipid membranes. Lipids, 38(4), 445-452. doi: 10.1007/s11745-003-1082-0

Gibb, Z., Lambourne, S. R., & Aitken, R. J. (2014). The paradoxical relationship between stallion fertility and oxidative stress. Biology of Reproduction, 91(3), 1-10. doi: 10.1095/biolreprod.114.118539

Giraud, M. N., Motta, C., Boucher, D., & Grizard, G. (2000). Membrane fluidity predicts the outcome of cryopreservation of human spermatozoa. Human Reproduction, 15(10), 2160-2164. doi: 10.1093/humrep/15.10.2160

Green, P., Glozman, S., & Yavin, E. (2001a). Ethyl docosahexaenoate-associated decrease in fetal brain lipid peroxide production is mediated by activation of prostanoid and nitric oxide pathways. Biochimica et Biophysica Acta, 1531(1-2), 156-164. doi: 10.1016/S1388-1981(01)00101-9

Green, P., Glozman, S., Weiner, L., & Yavin, E. (2001b). Enhanced free radical scavenging and decreased lipid peroxidation in the rat fetal brain after treatment with ethyl docosahexaenoate. Biochimica et Biophysica Acta, 1532(3), 203-212. doi: 10.1016/S1388-1981(01)00132-9

Harrison, R. A. P., & Vickers, S. E. (1990). Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. Journal of Reproduction and Fertility, 88(1), 343-352. doi: 10.1530/jrf.0.0880343

Howard, A. C., McNeil, A. K. & McNeil, P. L. (2011). Promotion of plasma membrane repair by vitamin E. Nature Communications, 20, 1-8. doi: 10.1038/ncomms1594

Hrudka, F. (1987). Cytochemical and ultracytochemical demonstration of cytochrome c oxidase in spermatozoa and dynamics of its changes accompanying ageing or induced by stress. International Journal of Andrology, 10, 809-828. doi: 10.1111/j.1365-2605.1987.tb00385.x

Kaeoket, K., Sang-Urai, P., Thamniyom, A., Chanapiwat, P., & Techakumphu, M. (2010). Effect of docosahexaenoic acid on quality of cryopreserved boar semen in different breeds. Reproduction in Domestic Animals, 45(3), 458-463. doi: 10.1111/j.1439-0531.2008.01239.x

Kaka, A., Haron, W., Yusoff, R., Yimer, N., Khumran, A. M., Sarsaifi, K., Ebrahimi, M. (2015). Effect of docosahexanoic acid on quality of frozen-thawed bull semen in BioXcell extender. Reproduction, Fertility and Development, 29(3), 490-495. doi: 10.1071/RD15089

Keshtgar, S., Fanaei, H., Bahmanpour, S., Azad, F., Ghannadi, A., & Kazeroni, M. (2012). In vitro effects of a-tocopherol on teratozoospermic semen samples. Andrologia, 44(1), 721-727. doi: 10.1111/j.1439-0272.2011.01256.x

Linfor, J. J., & Meyers, S. A. (2002). Detection of DNA damage in response to cooling injury in equine spermatozoa using single cell gel electrophoresis. Journal of Andrology, 23(1), 107-113. doi: 10.1002/j.1939-4640.2002.tb02603.x

Losano, J. D. A., Angrimani, D. S. R., Rui, B. R., Bicudo, L. C., Dalmazzo, A., Silva, B. C. S., Nichi, M. (2018). The addition of docosahexaenoic acid (DHA) and antioxidants (glutathione peroxidase and superoxide dismutase) in extenders to epididymal sperm cryopreservation in bulls. Zygote, 26(3), 199-206. doi: 10.1017/S0967199418000096

Love, C. C. (2005). The sperm chromatin structure assay: a review of clinical applications. Animal Reproduction Science, 89(1-4), 39-45. doi: 10.1016/j.anireprosci.2005.06.019

Nasiri, A. H., Towhidi, A., Zeinoaldini, S. (2012). Combined effect of DHA and alfa tocopherol supplementation during bull semen cryopreservation on sperm characteristics and fatty acid composition. Andrologia, 44(1), 550-555. doi: 10.1111/j.1439-0272.2011.01225.x

Naves, C. S., Beletti, M. E., Duarte, M. B., Vieira, R. C., Diniz, E. G., & Jacomini, J. O. (2004). Avaliação da cromatina espermática em equinos com azul de toluidina e acridine orange. Bioscience Journal, 20(3), 117-124.

Neill, A. R., & Masters, C. J. (1972). Metabolism of fatty acids by bovine spermatozoa. Biochemical Journal, 127(2), 375-385. doi: 10.1042/bj1270375

Nichi, M., Goovaerts, I. G. F., Cortada, C. N. M., Barnabe, V. H., De Clercq, J. B. P., & Bols, P. E. J. (2007). Roles of lipid peroxidation and cytoplasmic droplets on in vitro fertilization capacity of sperm collected from bovine epididymis stored at 4 and 34°C. Theriogenology, 67(2), 334-340. doi: 10.1016/j.theriogenology.2006.08.002

Nunes, D. B., Zorzatto, J. R., Costa e Silva, E. V., & Zúccari, C. E. S. N. (2008). Efficiency of short-term storage of equine semen in a simple design cooling system. Animal Reproduction Science, 104(2-4), 434-439. doi: 10.1016/j.anireprosci.2007.06.022

Sampaio, I. B. M. (2007). Estatística aplicada à experimentação animal (3a ed.) Belo Horizonte: Fundação de Ensino e Pesquisa em Medicina Veterinária e Zootecnia.

Sanocka, D., & Kurpisz, M. (2004). Reactive oxygen species and sperm cells. Reproductive Biology and Endocrinology, 2(12), 1-7. doi: 10.1186/1477-7827-2-12

Silva, D. M., Holden, S. A., Lyons, A., Souza, J. C., & Fair, S. (2017). In vitro addition of docosahexaenoic acid improves the quality of cooled but not frozen thawed stallion semen. Reproduction, Fertility and Development, 29(10), 2021-2027. doi: 10.1071/RD16473

Sindelar, P. J., Guan, Z., Dallner, G., & Ernster, L. (1999). The protective role of plasmalogens in iron induced lipid peroxidation. Free Radical Biology and Medicine, 26(3-4), 318-324. doi: 10.1016/s0891-5849(98)00221-4

Stillwell, W., & Wassall, S. R. (2003). Docosahexaenoic acid: membrane properties of a unique fatty acid. Chemistry and Physics of Lipids, 126(1), 1-27. doi: 10.1016/S0009-3084(03)00101-4

Takahashi, T., Itoh, R., Nishinomiya, H., Katoh, M., & Manabe, N. (2012). Effect of linoleic acid albumin in a dilution solution and long-term equilibration for freezing of bovine spermatozoa with poor freezability. Reproduction in Domestic Animals, 47(1), 92-97. doi: 10.1111/j.1439-0531.2011.01806.x

Towhidi, A., & Parks, J. E. (2012). Effect of n -3 fatty acids and alfa tocopherol on post thaw parameters and fatty acid composition of bovine sperm. Journal of Assisted Reproduction and Genetics, 29(10), 1051-1056. doi: 10.1007/s10815-012-9834-7

Tran, K., Wong, J. T., Lee, E., Chan, A. C., & Choy, P. C. (1996). Vitamin E potentiates arachidonate release and phospholipase A2 activity in rat heart myoblastic cells. Biochemical Journal, 319(2), 385-391. doi: 10.1042/bj3190385

Wassall, S. R., & Stillwell, W. (2009). Polyunsaturated fatty acid cholesterol interactions: domain formation in membranes. Biochimica et Biophysica Acta, 1788(1), 24-32. doi: 10.1016/j.bbamem.2008.10.011

Wayner, D. D. M., Burton, G. W., Ingold, K. U., Barclay, L. R. C., & Locke, S. J. (1987). The relative contributions of vitamin E, urate, ascorbate and proteins to the total peroxyl radical trapping antioxidant activity of human blood plasma. Biochimica et Biophysica Acta, 924(3), 408-419. doi: 10.1016/0304-4165(87)90155-3

Yavin, E. (2006). Docosahexaenoic acid: a pluripotent molecule acting as a membrane fluidizer, a cellular antioxidant and a modulator of gene expression. Nutrition and Health, 18(3), 261-262. doi: 10.1177/026010600601800308

Yoshida, K., Terao, J., Suzuki, T., & Takama, K. (1991). Inhibitory effect of phosphatidylserine on iron dependent lipid peroxidation. Biochemical and Biophysical Research Communications, 179(2), 1077-1081. doi: 10.1016/0006-291x(91)91929-7

Zerbinati, C., Caponecchia, L., Rago, R., Leoncini, E., Bottaccioli, A. G., Ciacciarelli, M., Iuliano, L. (2016). Fatty acids profiling reveals potential candidate markers of semen quality. Andrology, 4(6), 1094-1101. doi: 10.1111/andr.12236




DOI: http://dx.doi.org/10.5433/1679-0359.2020v41n1p167

Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433 / 1679-0359
E-mail:  semina.agrarias@uel.br
Este obra está licenciado com uma Licença  Creative Commons Atribuição-NãoComercial 4.0 Internacional