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Vertebrate reproductive science and technology
RESEARCH ARTICLE

The effect of the in vitro supplementation of exogenous long-chain fatty acids on bovine sperm cell function

M. Kiernan A , A. G. Fahey B and S. Fair A C
+ Author Affiliations
- Author Affiliations

A Department of Life Sciences, Faculty of Science and Engineering, Schrodinger Building, University of Limerick, Limerick, Ireland.

B School of Agriculture and Food Science, Veterinary Science Centre, University College Dublin, Belfield, Dublin 4, Ireland.

C Corresponding author. Email: sean.fair@ul.ie

Reproduction, Fertility and Development 25(6) 947-954 https://doi.org/10.1071/RD12204
Submitted: 26 June 2012  Accepted: 24 August 2012   Published: 5 October 2012

Abstract

This study aimed to investigate the effects of long-chain fatty acids (α-linolenic acid (ALA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), linoleic acid (LA), oleic acid (OA) and palmitic acid (PA)) at concentrations of 10–100 µM, on extended bull spermatozoa stored in vitro for up to 7 days. Progressive linear motion (PLM), viability (Experiments 1–3), ability to penetrate artificial mucus (Experiment 1), reactive oxygen species (ROS; Experiment 2) and superoxide production (Experiment 3) were assessed. Spermatozoa maintained the ability to penetrate artificial mucus up to Day 4, irrespective of treatment. In Experiments 2 and 3, DHA and EPA had detrimental effects on PLM and viability. PA preserved PLM and viability at levels greater than the control (P < 0.05), whilst keeping ROS levels to a minimum, particularly on Days 1 and 3 (P < 0.01) when ROS generation peaked in other treatments. In contrast, superoxide production peaked on Day 0 (Experiment 3) and declined thereafter with no significant effect of fatty acid. This study supports the notion that superoxide dominates on Day 0, whereas its breakdown products, hydrogen peroxide and the hydroxyl radical as assessed by CM-H2DCFDA, contribute to ROS generation on subsequent days.

Additional keywords : bull, motility, oxidative stress, reactive oxygen species, viability.


References

Aitken, R. J., and Baker, M. A. (2006). Oxidative stress, sperm survival and fertility control. Mol. Cell. Endocrinol. 250, 66–69.
Oxidative stress, sperm survival and fertility control.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XksVGjtrc%3D&md5=cec6f49d755c6ac1fd792001c5c92d33CAS | 16412557PubMed |

Aitken, R. J., Clarkson, J. S., and Fishel, S. (1989). Generation of reactive oxygen species, lipid peroxidation and human sperm function. Biol. Reprod. 41, 183–197.
Generation of reactive oxygen species, lipid peroxidation and human sperm function.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1MXlslWksLs%3D&md5=a41cdcbbfca11637745b7fa7f166315bCAS | 2553141PubMed |

Aitken, R. J., Wingate, J. K., De Iuliis, G. N., Koppers, A. J., and McLaughlin, E. A. (2006). Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa. J. Clin. Endocrinol. Metab. 91, 4154–4163.
Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XhtFSjtbvP&md5=1f9d3baf177ec44b7563075e80ec8f2aCAS | 16895947PubMed |

Aksoy, Y., Aksoy, H., Altinkaynak, K., Aydin, H. R., and Ozkan, A. (2006). Sperm fatty acid composition in subfertile men. Prostaglandins Leukot. Essent. Fatty Acids 75, 75–79.
Sperm fatty acid composition in subfertile men.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xosl2isrc%3D&md5=9b70d05ad7654d851a3c186c0d2a7a58CAS | 16893631PubMed |

Al Naib, A., Hanrahan, J. P., Lonergan, P., and Fair, S. (2011a). In vitro assessment of sperm from bulls of high and low field fertility. Theriogenology 76, 161–167.
In vitro assessment of sperm from bulls of high and low field fertility.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3Mrntlyjsg%3D%3D&md5=598dc6b33c964bb0ab7e07f770b93bb0CAS | 21396687PubMed |

Al Naib, A., Ward, F., Kelly, A., Wade, M., Marti, J., and Lonergan, P. (2011b). Effect of duration of storage at ambient temperature on fertilizing ability and mucus penetration ability of fresh bovine sperm. Theriogenology 76, 1070–1075.
Effect of duration of storage at ambient temperature on fertilizing ability and mucus penetration ability of fresh bovine sperm.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3MfgvFanug%3D%3D&md5=5b05f14889f4714c46c60b4450e710a8CAS | 21719089PubMed |

Alessandri, J. M., Guesnet, P., Vancassel, S., Astorg, P., Denis, I., Langelier, B., Aid, S., Poumes-Ballihaut, C., Champeil-Potokar, G., and Lavialle, M. (2004). Polyunsaturated fatty acids in the central nervous system: evolution of concepts and nutritional implications throughout life. Reprod. Nutr. Dev. 44, 509–538.
Polyunsaturated fatty acids in the central nervous system: evolution of concepts and nutritional implications throughout life.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXjtlelt7s%3D&md5=e7ace7124d013f9e6447e87f8460298bCAS | 15762297PubMed |

Alvarez, J. G., Touchstone, J. C., Blasco, L., and Storey, B. T. (1987). Spontaneous lipid peroxidation and production of hydrogen peroxide and superoxide in human spermatozoa. Superoxide dismutase as major enzyme protectant against oxygen toxicity. J. Androl. 8, 833–841.

Bønaa, K. H., Bjerve, K. S., Straume, B., Gram, I. T., and Thelle, D. (1990). Effect of eicosapentaenoic and docosahexaenoic acids on blood pressure in hypertension. N. Engl. J. Med. 322, 795–801.
Effect of eicosapentaenoic and docosahexaenoic acids on blood pressure in hypertension.Crossref | GoogleScholarGoogle Scholar | 2137901PubMed |

Box, G. E. P., and Cox, D. R. (1964). An analysis of transformations. J. R. Stat. Soc. Ser. B. Methodol. 26, 211–252.

Brenna, J. T., and Diau, G.-Y. (2007). The influence of dietary docosahexaenoic acid and arachidonic acid on central nervous system polyunsaturated fatty acid composition. Prostaglandins Leukot. Essent. Fatty Acids 77, 247–250.
The influence of dietary docosahexaenoic acid and arachidonic acid on central nervous system polyunsaturated fatty acid composition.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhsVWjtrvK&md5=f437633dbf77a548b074a0f5bfd0c3b8CAS | 18023566PubMed |

Castellano, C. A., Audet, I., Bailey, J., Laforest, J. P., and Matte, J. (2010). Dietary omega-3 fatty acids (fish oils) have limited effects on boar semen stored at 17°C or cryopreserved. Theriogenology 74, 1482–1490.
Dietary omega-3 fatty acids (fish oils) have limited effects on boar semen stored at 17°C or cryopreserved.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXht1Cls7%2FL&md5=2551643bc1ab0885774aa36b3cf82a52CAS | 20708242PubMed |

Childs, S., Hennessy, A., Sreenan, J., Wathes, D., Cheng, Z., Stanton, C., Diskin, M., and Kenny, D. (2008). Effect of level of dietary n-3 polyunsaturated fatty acid supplementation on systemic and tissue fatty acid concentrations and on selected reproductive variables in cattle. Theriogenology 70, 595–611.
Effect of level of dietary n-3 polyunsaturated fatty acid supplementation on systemic and tissue fatty acid concentrations and on selected reproductive variables in cattle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXpsVCrtbs%3D&md5=632552aafcfcb50c7cc7b0f24bdc8f76CAS | 18514298PubMed |

Cocco, T., Di, M., Papa, P. S., and Lorusso, M. (1999). Arachidonic acid interaction with the mitochondrial electron transport chain promotes reactive oxygen species generation. Free Radic. Biol. Med. 27, 51–59.
Arachidonic acid interaction with the mitochondrial electron transport chain promotes reactive oxygen species generation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXkvVKqsLw%3D&md5=c3e14b6ce415a5ad9a2a29eb929183d1CAS | 10443919PubMed |

Comhaire, F. H., and Mahmoud, A. (2003). The role of food supplements in the treatment of the infertile man. Reprod. Biomed. Online 7, 385–391.
The role of food supplements in the treatment of the infertile man.Crossref | GoogleScholarGoogle Scholar | 14656398PubMed |

Comhaire, F., Christophe, A., Zalata, A., Dhooge, W., Mahmoud, A., and Depuydt, C. (2000). The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men. Prostaglandins Leukot. Essent. Fatty Acids 63, 159–165.
The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXnslGisbc%3D&md5=048bb962981032aeea1a085182704ba5CAS | 10991774PubMed |

de Graaf, S. P., Peake, K., Maxwell, W. M. C., O’Brien, J. K., and Evans, G. (2007). Influence of supplementing diet with oleic and linoleic acid on the freezing ability and sex-sorting parameters of ram semen. Livest. Sci. 110, 166–173.
Influence of supplementing diet with oleic and linoleic acid on the freezing ability and sex-sorting parameters of ram semen.Crossref | GoogleScholarGoogle Scholar |

De Iuliis, G. N., Wingate, J. K., Koppers, A. J., McLaughlin, E. A., and Aitken, R. J. (2006). Definitive evidence for the non-mitochondrial production of superoxide anion by human spermatozoa. J. Clin. Endocrinol. Metab. 91, 1968–1975.
Definitive evidence for the non-mitochondrial production of superoxide anion by human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XkslCnu7Y%3D&md5=63cb694d068c2696ea08b6e2dec26254CAS | 16507629PubMed |

Gholami, H., Chamani, M., Towhidi, A., and Fazeli, M. (2010). Effect of feeding a docosahexaenoic acid-enriched nutriceutical on the quality of fresh and frozen–thawed semen in Holstein bulls. Theriogenology 74, 1548–1558.
Effect of feeding a docosahexaenoic acid-enriched nutriceutical on the quality of fresh and frozen–thawed semen in Holstein bulls.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhtlKhsLnF&md5=9d05265963403d3d50c7a838634223a9CAS | 20708237PubMed |

Hammiche, F., Vujkovic, M., Wijburg, W., De Vries, J. H. M., Macklon, N. S., Laven, J. S. E., and Steegers-Theunissen, R. P. M. (2010). Increased preconception omega-3 polyunsaturated fatty acid intake improves embryo morphology. Fertil. Steril. 95, 46–58.

Hossain, M. D. S., Tareq, K., Hammano, K. O. I., and Tsujii, H. (2007). Effect of fatty acids on boar sperm motility, viability and acrosome reaction. Reprod. Med. Biol. 6, 235–239.
Effect of fatty acids on boar sperm motility, viability and acrosome reaction.Crossref | GoogleScholarGoogle Scholar |

Kelso, K., Cerolini, S., Speake, B., Cavalchini, L., and Noble, R. (1997a). Effects of dietary supplementation with α-linolenic acid on the phospholipid fatty acid composition and quality of spermatozoa in cockerel from 24 to 72 weeks of age. J. Reprod. Fertil. 110, 53–59.
Effects of dietary supplementation with α-linolenic acid on the phospholipid fatty acid composition and quality of spermatozoa in cockerel from 24 to 72 weeks of age.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXksVansro%3D&md5=b4220957aadadb98c64bdb4752208249CAS | 9227357PubMed |

Kelso, K., Redpath, A., Noble, R., and Speake, B. (1997b). Lipid and antioxidant changes in spermatozoa and seminal plasma throughout the reproductive period of bulls. J. Reprod. Fertil. 109, 1.
| 1:CAS:528:DyaK2sXhs1Gnt70%3D&md5=bdb299351c6fdca04a9a6afd30f849f2CAS | 9068407PubMed |

Koppers, A. J., De Iuliis, G. N., Finnie, J. M., McLaughlin, E. A., and Aitken, R. J. (2008). Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa. J. Clin. Endocrinol. Metab. 93, 3199–3207.
Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXpvVOlsLw%3D&md5=4989e2013e46dffdb4eb19b233f3920aCAS | 18492763PubMed |

Koppers, A. J., Garg, M. L., and Aitken, R. J. (2010). Stimulation of mitochondrial reactive oxygen species production by un-esterified, unsaturated fatty acids in defective human spermatozoa. Free Radic. Biol. Med. 48, 112–119.
Stimulation of mitochondrial reactive oxygen species production by un-esterified, unsaturated fatty acids in defective human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhsFyku7rI&md5=ef15f07a29b8c836e00f5e39cc318ed3CAS | 19837155PubMed |

Kothari, S., Thompson, A., Agarwal, A., and du Plessis, S. S. (2010). Free radicals: their beneficial and detrimental effects on sperm function. Indian J. Exp. Biol. 48, 425–435.
| 1:CAS:528:DC%2BC3cXovV2rtb8%3D&md5=efd2e92de2d58bb838fe9d904d788d9bCAS | 20795359PubMed |

Lenzi, A., Picardo, M., Gandini, L., and Dondero, F. (1996). Lipids of the sperm plasma membrane: from polyunsaturated fatty acids considered as markers of sperm function to possible scavenger therapy. Hum. Reprod. Update 2, 246–256.
Lipids of the sperm plasma membrane: from polyunsaturated fatty acids considered as markers of sperm function to possible scavenger therapy.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXotVGgtLw%3D&md5=ce59faeab4db69aa4f59b450e162e9cdCAS | 9079417PubMed |

Lenzi, A., Gandini, L., Maresca, V., Rago, R., Sgro, P., Dondero, F., and Picardo, M. (2000). Fatty acid composition of spermatozoa and immature germ cells. Mol. Hum. Reprod. 6, 226–231.
Fatty acid composition of spermatozoa and immature germ cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXitVCgurc%3D&md5=dbe3b07d411c85ccf89c2d4e61911355CAS | 10694269PubMed |

Lukiw, W. J., and Bazan, N. G. (2008). Docosahexaenoic acid and the aging brain. J. Nutr. 138, 2510–2514.
Docosahexaenoic acid and the aging brain.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsVClsbrI&md5=21b46a22f68d9b1a86c45c7d64eba344CAS | 19022980PubMed |

Neill, A. R., and Masters, C. J. (1971). Incorporation of (U-14C)palmitic acid into the phospholipids of bovine semen. J. Reprod. Fertil. 24, 295–297.
Incorporation of (U-14C)palmitic acid into the phospholipids of bovine semen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE3MXktFOju7w%3D&md5=30b1cff874938d582f9be778c51e9317CAS | 4323937PubMed |

Oborna, I., Wojewodka, G., De Sanctis, J., Fingerova, H., Svobodova, M., Brezinova, J., Hajduch, M., Novotny, J., Radova, L., and Radzioch, D. (2010). Increased lipid peroxidation and abnormal fatty-acid profiles in seminal and blood plasma of normo-zoospermic males from infertile couples. Hum. Reprod. 25, 308–316.
Increased lipid peroxidation and abnormal fatty-acid profiles in seminal and blood plasma of normo-zoospermic males from infertile couples.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXntlWntw%3D%3D&md5=4c8051b96d0d042fc342566ea4ea1d33CAS | 19942612PubMed |

Ollero, M., Gil-Guzman, E., Lopez, M. C., Sharma, R. K., Agarwal, A., Larson, K., Evenson, D., Thomas, A. J., and Alvarez, J. G. (2001). Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility. Hum. Reprod. 16, 1912–1921.
Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXntlahu78%3D&md5=610c1bddacf4117c8a4a3679bdfcb1e7CAS | 11527898PubMed |

Rooke, J., Shao, C., and Speake, B. (2001). Effects of feeding tuna oil on the lipid composition of pig spermatozoa and in vitro characteristics of semen. Reproduction 121, 315–322.
Effects of feeding tuna oil on the lipid composition of pig spermatozoa and in vitro characteristics of semen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXhslSisbc%3D&md5=6894a05bd560520ee012038981601849CAS | 11226056PubMed |

Safarinejad, M. R., Hosseini, S. Y., Dadkhah, F., and Asgari, M. A. (2010). Relationship of omega-3 and omega-6 fatty acids with semen characteristics, and anti-oxidant status of seminal plasma: a comparison between fertile and infertile men. Clin. Nutr. 29, 100–105.
Relationship of omega-3 and omega-6 fatty acids with semen characteristics, and anti-oxidant status of seminal plasma: a comparison between fertile and infertile men.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhs1antLc%3D&md5=1479765dbf349155bc354e82d8c0fa3dCAS | 19666200PubMed |

Shannon, P. (1962). Effect of additions of volatile fatty acids on the viability and fertility of diluted bovine semen. Nature 196, 1225–1226.
Effect of additions of volatile fatty acids on the viability and fertility of diluted bovine semen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF3sXjsFajtQ%3D%3D&md5=fdd1efd01389a53915afcf12c2cf7a91CAS | 13976792PubMed |

Wathes, D. C., Abayasekara, D. R. E., and Aitken, R. J. (2007). Polyunsaturated fatty acids in male and female reproduction. Biol. Reprod. 77, 190–201.
Polyunsaturated fatty acids in male and female reproduction.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXot1OnsL0%3D&md5=2b573ad138ade4bd6481ad4d4866ef95CAS | 17442851PubMed |