Supplemental Glutathione Improves Post-Thaw Quality of Holstein Bulls Sperm in a Nanomicelle based Extender

Subject Areas : Camel

T. Nadri ¹ , A. Towhidi ² , S. Zeinoaldini ³ , G. Riazi ⁴ , M. Zhandi ⁵ , M. Sharafi ⁶

1 - Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
2 - Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
3 - Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
4 - Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran
5 - Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
6 - Department of Animal Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

Received: 2019-10-08 Accepted : 2019-11-30 Published : 2020-12-01

Keywords: Antioxidant, sperm, bovine, cryopreservation, nanomicelle,

Abstract :

Adding antioxidants to semen extenders could reduce post-thaw sperm damages. Glutathione is an important intracellular antioxidant that has free radicals scavenging properties and multiple effects on DNA and protein synthesis. In the present study, effects different levels of reduced glutathione (GSH) containing 0 (control), 1.0, 2.5, or 5.0 mM in a nanomicelle-based extender on post-thaw quality of bovine sperm were investigated. Initially, lecithin was added to a tris-based extender containing GSH. The mixture was then emulsified by slowly shaking and sonification so that nano size of particles was obtained. Thirty-six ejaculates from six Holstein bulls were used. The post-thaw quality parameters including computer-assisted sperm analysis of motility, plasma membrane functionality, membrane integrity, acrosome integrity, morphology and level of apoptosis were measured. Compared to the control medium, the total motility and membrane integrity were significantly better in samples containing 2.5 mM GSH (67.3±1.2% vs. 73±1.2% and 71.3±1.5% vs. 77.6±1.5%, respectively). The extender supplemented with 2.5 mM GSH had the highest percentage of viable sperm during apoptotic test. In conclusion, supplementation of lecithin nanomicelle-based extender with 2.5 mM GSH reduced damages to bovine sperm during cryopreservation.

References:

Agarwal A., Nallella K.P., Allamaneni S.S. and Said T.M. (2004). Role of antioxidants in treatment of male infertility: An overview of the literature. Reprod. Biomed. Online. 8, 616-627.

Alvarez J.G. and Storey B.T. (1992). Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a mode of sublethal cryodamage to human sperm during cryopreservation. J. Androl. 13, 232-241.

Anel-López L., Álvarez-Rodríguez M., García-Álvarez O., Álvarez M., Maroto-Morales A., Anel L., de Paz P., Garde J.J. and Martínez-Pastor F. (2012). Reduced glutathione and Trolox (vitamin E) as extender supplements in cryopreservation of red deer epididymal spermatozoa. Anim. Reprod. Sci. 135, 37-46.

Ansari M.S., Rakha B.A., Ullah N., Andrabi S.M.H., Iqbal S., Khalid M. and Akhter S. (2010). Effect of exogenous glutathione in extender on the freezability of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa. Anim. Sci. Pap. Rep. 28, 235-244.

Bachhawat A.K., Thakur A., Kaur J. and Zulkifli M. (2013). Glutathione transporters. Biochim. Biophys. Acta, Gen. Subj. 1830, 3154-3164.

Ballatori N., Krance S.M., Marchan R. and Hammond C.L. (2009). Plasma membrane glutathione transporters and their roles in cell physiology and pathophysiology. Mol. Asp. Med. 30, 13-28.

Bucak M.N., Ateşşahin A., Varışlı Ö., Yüce A., Tekin N. and Akçay A. (2007). The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen: microscopic and oxidative stress parameters after freeze–thawing process. Theriogenology. 67, 1060-1067.

Chatterjee S. and Gagnon C. (2001). Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol. Reprod. Dev. 59, 451-458.

Cohen R., Mukai C. and Travis A.J. (2016). Lipid regulation of acrosome exocytosis, Pp. 107-127 in Sperm Acrosome Biogenesis and Function during Fertilization. M.G. Buffone, Ed. Springer-Verlag Berlin Heidelberg Publisher, Germany.

De Oliveira R.A., Wolf C.A., de Oliveira Viu M.A. and Gambarini M.L. (2013). Addition of glutathione to an extender for frozen equine semen. J. Equine Vet. Sci. 33, 1148-1152.

Domínguez-Rebolledo Á.E., Fernández-Santos M.R., Bisbal A., Ros-Santaella J.L., Ramón M., Carmona M., Martínez-Pastor F. and Garde J.J. (2010). Improving the effect of incubation and oxidative stress on thawed spermatozoa from red deer by using different antioxidant treatments. Reprod. Fertil. Dev. 22, 856-870.

Estrada E., del Álamo M.M.R., Rodríguez-Gil J.E. and Yeste M. (2017). The addition of reduced glutathione to cryopreservation media induces changes in the structure of motile subpopulations of frozen-thawed boar sperm. Cryobiology. 78, 56-64.

Ferreira G., Costa C., Bassaizteguy V., Santos M., Cardozo R., Montes J., Settineri R. and Nicolson G.L. (2018). Incubation of human sperm with micelles made from glycerophospholipid mixtures increases sperm motility and resistance to oxidative stress. PloS One. 13, e0197897.

Gadea J., Molla M., Selles E., Marco M., Garcia-Vazquez F. and Gardon J. (2011). Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology. 62, 40-46.

Gadea J., Sellés E., Marco M.A., Coy P., Matás C., Romar R. and Ruiz S. (2004). Decrease in glutathione content in boar sperm after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Theriogenology. 62, 690-701.

Geva E., Lessing J.B., Lerner-Geva L. and Amit A., (1998). Free radicals, antioxidants and human spermatozoa: clinical implications. Hum. Reprod. 13, 1422-1424.

Gil J., Lundeheim N., Söderquist L. and Rodrı́guez-Martı́nez H. (2003). Influence of extender, temperature, and addition of glycerol on post-thaw sperm parameters in ram semen. Theriogenology. 59, 1241-1255.

Guthrie H. and Welch G. (2012). Effects of reactive oxygen species on sperm function. Theriogenology. 78, 1700-1708.

Mata-Campuzano M., Álvarez-Rodríguez M., Álvarez M., Tamayo-Canul J., Anel L., de Paz P. and Martínez-Pastor F. (2015). Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender. Theriogenology. 83, 520-528.

Mata-Campuzano M., Álvarez-Rodríguez M., Tamayo-Canul J., López-Urueña E., de Paz P., Anel L., Martínez-Pastor F. and Álvarez M. (2014). Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants: Effect of temperature, extender and storage time. Anim. Reprod. Sci. 151, 137-147.

Munsi M., Bhuiyan M., Majumder S. and Alam M. (2007). Effects of exogenous glutathione on the quality of chilled bull semen. Reprod. Dom. Anim. 42, 358-362.

Nadri T., Towhidi A., Zeinoaldini S., Martínez-Pastor F., Mousavi M., Noei R., Tar M. and Sangcheshmeh A.M. (2019). Lecithin nanoparticles enhance the cryosurvival of caprine sperm. Theriogenology. 133, 38-44.

Najafi A., Taheri R.A., Mehdipour M., Farnoosh G. and Martínez-Pastor F. (2018). Lycopene-loaded nanoliposomes improve the performance of a modified Beltsville extender broiler breeder roosters. Anim. Reprod. Sci. 195, 168-175.

Najafi A., Zhandi M., Towhidi A., Sharafi M., Sharif A.A., Motlagh M.K. and Martinez-Pastor F. (2013b). Trehalose and glycerol have a dose-dependent synergistic effect on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender. Cryobiology. 66, 275-282.

Niki E. (2014). Antioxidants: Basic principles, emerging concepts, and problems. Biomed. J. 37, 106-201.

Ogata K., Sasaki A., Kato Y., Takeda A., Wakabayashi M., Sarentonglaga B., Yamaguchi M., Hara A., Fukumori R. and Nagao Y. (2015). Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination. J. Reprod. Dev. 61, 116-122.

Rawash Z.M., Ibrahim E.M. and El-Raey M. (2018). Effects of reduced glutathione on Boer goat semen freezability. Asian Pac. J. Reprod. 7, 33-38.

Richens J.L., Tyler A.I., Barriga H.M., Bramble J.P., Law R.V., Brooks N.J., Seddon J.M., Ces O. and O’Shea P. (2017). Spontaneous charged lipid transfer between lipid vesicles. Sci. Repod. 7, 12606-12610.

Salmani H., Towhidi A., Zhandi M., Bahreini M. and Sharafi M. (2014). In vitro assessment of soybean lecithin and egg yolk based diluents for cryopreservation of goat semen. Cryobiology. 68, 276-280.

SAS Institute. (2002). SAS^®/STAT Software, Release 9.1. SAS Institute, Inc., Cary, NC. USA.

Schäfer S. and Holzmann A. (2000). The use of transmigration and Spermac™ stain to evaluate epididymal cat spermatozoa. Anim. Reprod. Sci. 59, 201-211.

Sharafi M., Forouzanfar M., Hosseini S.M., Hajian M., Ostadhosseini S., Hosseini L., Abedi P., Nili N., Rahmani H.R., Reza Javaheri A. (2009). In vitro comparison of soybean lecithin based-extender with commercially available extender for ram semen cryopreservation. Int. J. Fertil. Steril. 3, 149-152.

Sharafi M., Zhandi M., Shahverdi A. and Shakeri M. (2015). Beneficial effects of nitric oxide induced mild oxidative stress on post-thawed bull semen quality. Int. J. Fertil. Steril. 9, 230-237.

Sinha M., Sinha A., Singh B. and Prasad R. (1996). The effect of glutathione on the motility, enzyme leakage and fertility of frozen goat semen. Anim. Reprod. Sci. 41, 237-243.

Thys M., Nauwynck H., Maes D., Hoogewijs M., Vercauteren D., Rijsselaere T., Favoreel H. and Van Soom A. (2009). Expression and putative function of fibronectin and its receptor (integrin α5β1) in male and female gametes during bovine fertilization in vitro. Reproduction. 138, 471-482.

Triwulanningsih E., Situmorang P., Sugiarti T., Sianturi R. and Kusumaningrum D. (2010). Effect of glutathione addition to the sperm diluent medium on quality of bovine chilled semen. Indonesian J. Agric. 3, 60-65.

Ugur M.R., Saber Abdelrahman A., Evans H.C., Gilmore A.A., Hitit M., Arifiantini R.I., Purwantara B., Kaya A. and Memili E. (2019).Advances in cryopreservation of bull sperm. Front. Vet. Sci. 6,268-275.

Varghese A., Das S., Bhattacharya A., Bhattacharya S., Mandal M. and Agarwal A. (2005). Effect of cryoprotective additives-reduced glutathione, acetyl-L-carnitine on sperm membrane lipid peroxidation, DNA integrity and recovery of motile human sperm. Fertil. Steril. 84, 410-411.

Wachter A., Wolf S., Steininger H., Bogs J. and Rausch T. (2005). Differential targeting of GSH1 and GSH2 is achieved by multiple transcription initiation: Implications for the compartmentation of glutathione biosynthesis in the Brassicaceae. Plant. J. 41, 15-30.

Wang A.W., Zhang H., Ikemoto I., Anderson D.J. and Loughlin K.R. (1997). Reactive oxygen species generation by seminal cells during cryopreservation. Urology. 49, 921-925.

Zhandi M. and Ghadimi V. (2014). Effect of glutathione-supplemented INRA82 extender on miniature Caspian stallion sperm quality during storage at 5 C. J. Equine. Vet. Sci. 34, 606-610.

Zieliński R., Ikeda S., Nomura H. and Kato S. (1989). Effect of temperature on micelle formation in aqueous solutions of alkyltrimethylammonium bromides. J. Colloid Interface Sci. 129, 175-184.