Animal Reproduction (AR)
Animal Reproduction (AR)

Coenzyme Q-10 improves preservation of mitochondrial functionality and actin structure of cryopreserved stallion sperm

Renata Lançoni; Eneiva Carla Carvalho Celeghini; Valdemar De Giuli Júnior; Carla Patricia Teodoro de Carvalho; Gabriela Bertaiolli Zoca; Laura Nataly Garcia-Oliveros; Leonardo Batissaco; Letícia Zoccolaro Oliveira; Rubens Paes de Arruda

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Abstract: Coenzyme Q-10 (CoQ-10) is a cofactor for mitochondrial electron transport chain and may be an alternative to improve sperm quality of cryopreserved equine semen. This work aimed to improve stallion semen quality after freezing by adding CoQ-10 to the cryopreservation protocol. Seven saddle stallions were utilized. Each animal was submitted to five semen collections and freezing procedures. For cryopreservation, each ejaculate was divided in three treatments: 1) Botucrio® diluent (control); 2) 50 μmol CoQ-10 added to Botucrio® diluent; 3) 1 mmol CoQ-10 added to Botucrio® diluent. Semen batches were analyzed for sperm motility characteristics (CASA), plasma and acrosomal membranes integrity and mitochondrial membrane potential (by fluorescence probes propidium iodide, Hoechst 33342, FITC-PSA and JC-1, respectively), alterations in cytoskeletal actin (phalloidin-FITC) and mitochondrial function (diaminobenzidine; DAB). The 1 mmol CoQ-10 treatment presented higher (P<0.05) amount (66.8%) of sperm cells with fully stained midpiece (indicating high mitochondrial activity) and higher (P<0.05) amount (81.6%) of cells without actin reorganization to the post-acrosomal region compared to control group (60.8% and 76.0%, respectively). It was concluded that the addition of 1 mmol CoQ-10 to the freezing diluent was more effective in preserving mitochondria functionality and cytoskeleton of sperm cells submitted to cryopreservation process.


antioxidant, cytoskeleton, cryopreservation, equine, sperm quality


Affonso FJ, Carvalho HF, Lançoni R, Lemes KM, Leite TG, Oliveira LZ, Celeghini ECC, de Arruda RP. Addition of antioxidants myoinositol, ferulic acid, and melatonin and their effects on sperm motility, membrane integrity, and reactive oxygen species production in cooled equine semen. J Equine Vet Sci. 2017;59:57-63.

Al-Essawe EM, Johannisson A, Wulf M, Aurich C, Morrell JM. Cryobiology Improved cryosurvival of stallion spermatozoa after colloid centrifugation is independent of the addition of seminal plasma. Cryobiology. 2018;81:145-52. PMid:29397923.

Allen WR, Wilsher S. The influence of mare numbers, ejaculation frequency and month on the fertility of Thoroughbred stallions. Equine Vet J. 2011;44(5):535-41. PMid:22168381.

Alleva R, Scararmucci A, Mantero F, Bompadre S, Leoni L, Littarru GP. The protective role of ubiquinol-10 against formation of lipid hydroperoxides in human seminal fluid. Mol Aspects Med. 1997;18(Suppl):S221-8. PMid:9266525.

Balercia G, Mancini A, Paggi F, Tiano L, Pontecorvi A, Boscaro M, Lenzi A, Littarru GP. Coenzyme Q 10 and male infertility. J Endocrinol Invest. 2009;32(7):626-32. PMid:19509475.

Bentinger M, Brismar K, Dallner G. The antioxidant role of coenzyme Q. Mitochondrion. 2007;7(Suppl):S41-50. PMid:17482888.

Brener E, Rubinstein S, Cohen G, Shternall K, Rivlin J, Breitbart H. Remodeling of the actin cytoskeleton during mammalian sperm capacitation and acrosome reaction. Biol Reprod. 2003;68(3):837-45. PMid:12604633.

Celeghini E, Arruda R, de Andrade A, Nascimento J, Raphael C. Practical techniques for bovine sperm simultaneous fluorimetric assessment of plasma, acrosomal and mitochondrial membranes. Reprod Domest Anim. 2007;42(5):479-88. PMid:17845603.

Correa LM, Thomas A, Meyers SA. The macaque sperm actin cytoskeleton reorganizes in response to osmotic stress and contributes to morphological defects and decreased motility. Biol Reprod. 2007;77(6):942-53. PMid:17823088.

Dai J, Zhan C, Xu W, Wang Z, Nie D, Zhao X, Zhang D, Gu Y, Wang L, Chen Z, Qiao Z. Nicotine elevates sperm motility and induces Pfn1 promoter hypomethylation in mouse testis. Andrology. 2015;3(5):967-78. PMid:26311342.

Eddy EM, Toshimori K, O’Brien DA. Fibrous sheath of mammalian spermatozoa. Microsc Res Tech. 2003;61(1):103-15. PMid:12672126.

Fawcett DW. The mammalian spermatozoon. Dev Biol. 1975;44(2):394-436. PMid:805734.

Flores E, Fernández-novell JM, Peña A, Rigau T, Rodríguez-gil JE. Cryopreservation-induced alterations in boar spermatozoa mitochondrial function are related to changes in the expression and location of midpiece mitofusin-2 and actin network. Theriogenology. 2010;74(3):354-63. PMid:20416937.

Gibb Z, Lambourne SR, Aitken RJ, Al GET. The paradoxical relationship between stallion fertility and oxidative stress. Biol Reprod. 2014;91(3):1-10. PMid:25078685.

Hall SM, Evans J, Haworth SG. Influence of cold preservation on the cytoskeleton of cultured pulmonary arterial endothelial cells. Am J Respir Cell Mol Biol. 1993;9(14):106-14. PMid:8338672.

Holmes KC, Popp D, Gebhard W, Kabsch W. Atomic model of the actin filament. Nature. 1990;347(6288):44-9. PMid:2395461.

Hrudka F. Cytochemical and ultracytochemical demonstration of cytochrome c oxidase in spermatozoa and dynamics of its changes accompanying ageing or induced by stress. Int J Androl. 1987;10(6):809-28. PMid:2828243.

Lançoni R, Celeghini ECC, Alves MBR, Lemes KM, Gonella-Diaza AM, Oliveira LZ, Arruda RP. Melatonin added to cryopreservation extenders improves the mitochondrial membrane potential of postthawed equine sperm. J Equine Vet Sci. 2018;69:78-83.

Lewin A, Lavon H. The effect of coenzyme Q-10 on sperm motility and function. Mol Aspects Med. 1997;18(Suppl):S213-9. PMid:9266524.

Lin M, Hess R, Aitken RJ. Induction of sperm maturation in vitro in epididymal cell cultures of the tammar wallaby (Macropus eugenii): disruption of motility initiation and sperm morphogenesis by inhibition of actin polymerization. Reproduction. 2002;124(1):107-17. PMid:12090924.

Littarru GP, Tiano L. Clinical aspects of coenzyme Q 10 : an update. Nutrition. 2010;26(3):250-4. PMid:19932599.

Liu DY, Clarke GN, Baker HWG. Exposure of actin on the surface of the human sperm head during in vitro culture relates to sperm morphology, capacitation and zona binding. Hum Reprod. 2005;20(4):999-1005. PMid:15650044.

Macías García B, Ortega Ferrusola C, Aparicio IM, Miró-Morán A, Morillo Rodriguez A, Gallardo Bolaños JM, González Fernández L, Balao da Silva CM, Rodríguez Martínez H, Tapia JA, Peña FJ. Toxicity of glycerol for the stallion spermatozoa : effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential. Theriogenology. 2012;77(7):1280-9. PMid:22326587.

Nascimento J, Raphael CF, Andrade AFC, Alonso MA, Celeghini ECC, Arruda RP. Effects of sperm concentration and straw volume on motion characteristics and plasma, acrosomal, and mitochondrial membranes of equine cryopreserved spermatozoa. J Equine Vet Sci. 2008;28(6):351-8.

Ortega-Ferrusola C, Fernández LG, Sandoval CS, García BM, Martínez HR, Tapia JA, Peña FJ. Inhibition of the mitochondrial permeability transition pore reduces “ apoptosis like ” changes during cryopreservation of stallion spermatozoa. Theriogenology. 2010;74(3):458-65. PMid:20451990.

Peña FJ, Plaza Davila M, Ball BA, Squires EL, Martin Muñoz P, Ortega Ferrusola C, Balao da Silva C. The impact of reproductive technologies on stallion mitochondrial function. Reprod Domest Anim. 2015;537(4):529-37.

Rossi M, Falomo ME, Mantovani R, Rossi M, Falomo ME, Mantovani R. Role of coenzyme Q and vitamin E on stallion semen motility evaluated both in frozen and cooled-stored semen. Ital J Anim Sci. 2016;15(4):595-603.

Tsunoda RH, Arruda RP, Serrano-Recalde EC, Oliveira BMM, Florez-Rodriguez SA, Alves MBR, Lançoni R, Nichi M, Celeghini ECC. Addition of pentoxifylline to skim milk-based extender on frozen-thawed equine sperm. J Equine Vet Sci. 2015;35(10):823-9.

Turunen M, Olsson J, Dallner G. Metabolism and function of coenzyme Q. Biochim Biophys Acta. 2004;1660(1-2):171-99. PMid:14757233.

Watson PF. Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod Fertil Dev. 1995;7(4):871-91. PMid:8711221.

Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci. 2000;60-61:481-92. PMid:10844218.

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