OLGA BÜYÜKLEBLEBİCİ, SERHAT BÜYÜKLEBLEBİCİ, PÜRHAN BARBAROS TUNCER, TANER ÖZGÜRTAŞ, UMUT TAŞDEMİR

Effect of ?-Lipoic Acid Supplementation to Extender on Quality of Frozen-Thawed Bull Semen

Bu çalışma dondurma-çözündürme işleminin neden olduğu oksidadif stresden dolayı boğa spermasında meydane gelen hasarlar üzerine sulandırıcıya katılan ?-lipoik asidin (ALA) muhtemel koruyucu etkisini araştırmak amacı ile yapıldı. Ejakülatler haftada bir kez olmak üzere suni vajen yardımı ile toplandı, sonrasında beş eşit parçaya ayrıldı ve kontrol grubu dışında diğerleri farklı dozlarda ALA içeren Tris bazlı sperma sulandırıcısı ile sulandırıldı. Sulandırılan spermalar 4°C'de 4 saat süre ile ekülibre edildi ve otomatik sperma dondurma cihazı kullanılarak donduruldu. Dondurulan spermalar çözündürülerek plazma membran bütünlüğünün yanı sıra spermanın ileri yönlü hareketi ve sperma hareket özellikleri değerlendirildi. Biyokimyasal analizler ticari kit kullanılarak spektrofotometre de yapıldı. ALA0.5 grubu dışında ALA ilave edilmiş sulandırıcı ile sulandırılan gruplarda dondurma çözündürme sonrası CASA ve ileri yönlü spermatozoa hareketi ile VAP, VSL, VCL (spermatozoonun tüm hareketlerinin ortalaması, spermatozonun ilk harekete başladığı ve hareketini sonlandırdığı yer arasındaki en kısa mesafe, gerçek eğrisel yolda kat ettiği mesafe) gibi spermatozoa hareket özellikleri üzerine olumsuz etki gösterdi. Bunun yanında, kontrol grubu ile karşılaştırıldığında ALA0.5 akrozom ve total anormalitenin daha düşük oranlarda olmasını sağladı. Sonuç olarak, bu çalışmada elde edilen bulgular Tris bazlı sperma sulandırıcısına ilave edilen ALA0.5'in morfoloji ve plazma membran bütünlüğü üzerine olumlu etkilerinin olduğunu gösterdi

Sulandırıcıya Katılan ?-Lipoik Asidin Dondurulmuş-Çözdürülmüş Boğa Spermasının Kalitesi Üzerine Etkisi

This study was conducted to investigate the possible protective effect of ?-lipoic acid (ALA) supplementation to extender on damages in the quality of bull semen elicited by freze-thawing process-induced oxidative stres. Ejaculates were collected via an artificial vagina from the bulls at once a week. Then they were split into five aliquots and extended with the Tris base extender containing different doses of ALA, except control. The extended samples were equilibrated slowly to 4°C for 4 h and then frozen using a digital freezing machine. Frozen straws were thawed to analyse progressive motility and sperm motility characteristics as well as plasma membrane integrity. Biochemical assays were performed in a spectrophotometer using commercial kits. Treatment of extender with ALA groups caused a significant decrease in percentages of post-thaw sperm CASA, progressive motilities and sperm motility characteristics such as VAP, VSL and VCL except ALA0.5. Besides ALA0.5 showed lower percentages of acrosome and total abnormalities in comparison to the control. In conclusion, findings generated here showed that ALA0.5 supplementation in Tris based semen extender was of great beneficial effect on frozen-thawed bull semen in terms of morphology and plasma membrane integrity

PDF

___

Watson PF: Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod Fertil Develop, 7, 871-891, 1995. DOI: 10.1071/RD9950871
Üstüner B, Alçay S, Nur Z, Sağırkaya H, Soylu MK: Effect of egg yolk and soybean lecithin on tris-based extender in post-thaw ram semen quality and in vitro fertility. Kafkas Univ Vet Fak Derg, 20, 393-398, 2014. DOI: 9775/kvfd.2013.10248
Wang AW, Zhang H, Ikemoto I, Anderson DJ, Loughlin KR: Reactive oxygen species generation by seminal cells during cryopreservation. Urology, 49, 921-925, 1997. DOI: 10.1016/S0090-4295(97)00070-8
Vernet P, Aitken RJ, Drevet JR: Antioxidant strategies in the epididymis. Mol Cell Endocrinol, 216, 31-39, 2004. DOI: 10.1016/j.mce.2003.10.069
Aitken RJ, Clarkson JS, Fishel S: Generation of reactive oxygen species, lipid peroxidation, and human sperm function. Biol Reprod, 41, 197, 1989.
Drevet JR: The antioxidant glutathione peroxidase family and spermatozoa: A complex story. Mol Cell Endocrinol, 250, 70-79, 2006. DOI: 1016/j.mce.2005.12.027
Ross C, Morriss A, Khairy M, Khalaf Y, Braude P, Coomarasamy A, El-Toukhy TA: A systematic review of the effect of oral antioxidants on male infertility. Reprod BioMed Online, 20, 711-723, 2010. DOI: 10.1016/j. rbmo.2010.03.008
Lester P, Witt EH, Tritschler HJ: Alpha-lipoic acid as a biological antioxidant. Free Rad Biol Med, 19, 227-250, 1995. DOI: 10.1016/0891- (95)00017-R
Devasagayam TPA: Antioxidant activity in sugarcane juice and its protective role against radiation induced DNA damage. Bioch Bioph Acta, 876, 507-514, 1986. DOI: 10.1016/j.foodchem.2007.07.066
Habig WH, Pabst MJ, Jakoby WB: Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem, 249, 7139, 1974.
Suzuki YJ, Tsuchiya M, Packer L: Thioctic acid and dihydrolipoic acid are novel antioxidants which interact with reactive oxygen species. Free Radic Res Commun 15, 255-263, 1991. DOI: 10.3109/10715769109105221
Bustamante J, Lodge JK, Marcocci L, Tritschler HJ, Packer L, Rihn BH: Alpha-lipoic acid in liver metabolism and disease. Free Rad Biol Med, , 1023-1039, 1998. DOI: 10.1016/S0891-5849(97)00371-7
Revel SG, Mrode RA: An osmotic resistance test for bovine semen. Anim Reprod Sci, 36, 77-86, 1994. DOI: 10.1016/0378-4320(94)90055-8
Schafer S, Holzman A: The use of transmigration and Spermac stain to evaluate epididymal cat spermatozoa. Anim Reprod Sci, 59, 201-211, 2000. DOI: 10.1016/S0378-4320(00)00073-7
Taşdemir U, Tuncer PB, Buyukleblebici S, Ozgurtas T, Durmaz E, Buyukleblebici O: Effects of various antioxidants on cryopreserved bull sperm quality. Kafkas Univ Vet Fak Derg, 20, 253-258, 2014. DOI: 10.9775/ kvfd.2013.9964
Baumer J, Ball BA, Linfor JJ: Assessment of the cryopreservation of equine spermatozoa in the presence of enzyme scavengers and antioxidants. Am J Vet Res, 66, 772-779, 2005. DOI: 0.2460/ajvr.2005.66.772
Kasamo K, Kagita F, Yamanishi H, Sakaki T: Low temperature induced changes in the thermotropic properties and fatty acid composition of the plasma membrane and tonoplast and cultured rice (Oryza sativa L.). Plant Cell Physiol, 33, 609-616, 1992.
Kagan VE, Shvedova A, Serbinova E, Khan S, Swanson C, Powell R, Packer L: Dihydrolipoic acid a universal antioxidant both in the membrane and in the aqueous phase. Biochem Pharmacol, 44, 1637-1649, DOI: 10.1016/0006-2952(92)90482-X
Selvakumar E, Prahalathan C, Sudharsan PT, Varalakshmi P: Chemoprotective effect of lipoic acid against cyclophosphamide-induced changes in the rat sperm. Toxicology, 217, 71-78, 2006. DOI: 10.1016/j. tox.2005.08.020
Ma H, Quan F, Chen D, Zheng Y, Zhang B, Wang Y, Zhang Y: Protective function of alpha-lipoic acid on sperm motility and mitochondrial function during goat sperm-mediated gene transfer. Small Rumin Res, , 191-198, 2011. DOI: 10.1016/j.smallrumres.2011.04.006
Chatterjee S, Gagnon C: Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Develop, 59, 451-458, 2001. DOI: 10.1002/mrd.1052
Büyükleblebici S, Taşdemir U, Tuncer PB, Durmaz E, Özgürtaş T, Büyükleblebici O, Coşkun E, Gürcan İS: Can linoleic acid improve the quality of frozen-thawed bull sperm? Cryoletters, 35 (6): 473-481, 2014.
Soares MP, Brandelli A, Celeghini ECC, Arruda RP, Rodriguez SAF: Effect of cis-9, trans-11 and trans-10,cis-12 isomers of conjugated linoleic acid on the integrity and functionality of cryopreserved bovine spermatozoa. Cryobiology, 67, 102-105, 2013. DOI: 10.1016/j. cryobiol.2013.05.003
Castellano CA, Audeta I, Bailey JI, Laforest JP, Matte JJ: Dietary omega-3 fatty acids (fish oils) have limited effects on boar semen stored at 17°C or cryopreserved. Theriogenology. 74, 1482-1490, 2010. DOI: 1016/j.theriogenology.2010.06.020
Gliozzi TM, Zaniboni L, Maldjian A, Luzi F, Maertens L, Cerolini S: Quality and lipid composition of spermatozoa in rabbits fed DHA and vitamin E rich diets. Theriogenology, 71, 910-919, 2009. DOI: 10.1016/j. theriogenology.2008.10.022
Grady S, Cavinder C, Brinsko S, Forrest D, Sawyer J, Scott B: Dietary supplementation of two varying sources of n-3 fatty acids and subsequent effects on fresh, cooled and frozen seminal characteristics of stallions. Prof Anim Sci, 25, 768-773, 2009.
Takahashi T, Itoh R, Nishinomiya H, Katoh M, Manabe N: Effect of linoleic acid albumin in a dilution solution and long-term equilibration for freezing of bovine spermatozoa with poor freezability. Reprod Domest Anim, 42, 92-97, 2012. DOI: 10.1111/j.1439-0531.2011.01806.x
Tuncer PB, Sariozkan S, Bucak MN, Ulutas PA, Akalın PP, Buyukleblebici S, Canturk F: Effect of glutamine and sugars after bull spermatozoa cryopreservation. Theriogenology, 75, 1459-1465, 2011. DOI: 10.1016/j.theriogenology.2010.12.006
Bucak MN, Baspınar N, Tuncer PB, Coyan K, Sariozkan S, Akalın PP, S. Buyukleblebici S, Kuçukgunay S: Effects of curcumin and dithioerythritol on frozen-thawed bovine semen. Andrologia, 44, 102- , 2012. DOI: 10.1111/j.1439-0272.2010.01146.x
Graham JK, Foote RH: Effect of several lipids, fatty acyl chain length, and degree of unsaturation on the motility of bull spermatozoa after cold shock and freezing. Cryobiology, 24, 42-52, 1987. DOI: 10.1016/0011- (87)90005-8
Vassallo-Agius R, Watanabe T, Yoshizaki G, Satoh S, Takeuchi Y: Quality of eggs and spermatozoa of rainbow trout fed an n-3 essential fatty acid-deficient diet and its effects on the lipid and fatty acid components of eggs, semen and livers. Fisheries Sci, 67, 818-827, 2001. DOI: 10.1046/j.1444- 2001.00328.x
Michael A, Alexopoulos C, Pontiki E, Hadjipavlou-Litina D, Saratsis P, Boscos C: Effect of antioxidant supplementation on semen quality and reactive oxygen species of frozen-thawed canine spermatozoa. Theriogenology, 68, 204-212, 2007. DOI: 10.1016/j. theriogenology.2007.04.053
Bilodeau JF, Blanchette S, Gagnon C, Sirard MA: Thiols prevent H2O2-mediated loss of sperm motility in cryopreserved bull semen. Theriogenology, 56, 275-286, 2001. DOI: 10.1016/S0093-691X(01)00562-3
Bucak MN, Tuncer PB, Sarıozkan S, Ulutas PA, Coyan K, Baspınar N, Ozkalp B: Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen. Res Vet Sci, 87, 468-478, 2009. DOI: 10.1016/j.rvsc.2009.04.014
Kulaksız R, Bucak MN, Akçay E, Sakin F, Daşkın A, Ateşşahin A: The effects of different extenders and myo-Inositol on post-thaw quality of ram semen. Kafkas Univ Vet Fak Derg, 17, 217-222, 2011. DOI: 10.9775/ kvfd.2010.3024