Rat Epididimal Spermasının Kısa Süreli Saklanmasında Farklı Sıcaklık Dereceleri ve Karnitinin Etkileri

Yıl 2019, Cilt: 16 Sayı: 1, 30 - 37, 25.03.2019

Serpil Sarıözkan

https://doi.org/10.32707/ercivet.538016

Öz

Bu çalışmanın amacı, kısa süreli saklanan
(0., 6., 12., 24. ve 48. saat) rat epididimal spermasında farklı saklama
derecelerinin (4 ^oC ve 37 ^oC) ve karnitinin spermatolojik
parametreler üzerine etkisini araştırmaktır. Araştırmada sperma donörü
olarak 10 adet 8-10 haftalık yaş ve 200-250 g ağırlıklarında Wistar-Albino erişkin rat
kullanıldı. Epididimisler anestezi altındaki ratlardan cerrahi teknikle alındı.
Epididimal sperma numuneleri önce 4 ayrı gruba ayrıldı, numuneler Tris (T) ve
Tris + 2 mM  karnitin (TC) ile
sulandırıldı. Ardından sulandırılan sperma numuneleri 4 ^oC (T - TC)
ve 37 ^oC (T - TC) de 48 saat süreyle kısa süreli saklandı. Saklama
süresinin ardından motilite ve anormal spermatozoa oranı analiz edildi.
Uygulama 10 kez tekrar edildi. Çalışmanın sonucunda, tüm deneysel gruplarda 48.
saat sonunda anormal spermatozoa oranında artış ve motilite oranında düşme
saptandı. Her iki sıcaklık derecesinde motilite ve spermatozoa morfolojik
yapısının karnitin katılan gruplarda (TC) daha iyi korunduğu belirlendi. Sonuç
olarak, 4 ^oC deki motilite oranı 37 ^oC ye kıyasla daha
yüksek ve anormal spermatozoa oranı ise daha düşük bulundu. 

Anahtar Kelimeler

Rat, epididimal sperma, karnitin, kısa süreli saklama, spermatolojik parametreler

Kaynakça

• 1. Agarwal A, Virk G, Ong C, Du Plessis SS. Effect of oxidative stress on male reproduction. World J Mens Health 2014; 32(1): 1-17. 2. Ağca Y, Critser JK. Assisted reproductive technologies and genetic engineering in rats, Suckow MA, Weisbroth SH, Franklin CL. (eds). In: The Laboratory Rat. San Diego: Academic Press, 2005; pp.165-90. 3. Aitken RJ, Clarkson JS, Fishel S. Generation of reactive oxygen species, lipid peroxidation and human sperm function. Biol Reprod 1989; 41: 183-97. 4. Aitken RJ. Free radicals, lipid peroxidation and sperm function. Reprod Fertil Dev 1995; 7(4): 659-68. 5. Audrins P, Holden CA, McLachlan RI, Kovacs GT. Semen storage for special purposes at monash IVF from 1977 to 1997. Fertil Steril 1999; 72(1): 179-81. 6. Bailey JL, Bilodeau JF, Cormier N. Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. J Androl 2000; 21(1): 1-7. 7. Beconi MT, Affranchino MA, Schang LM, Beorlegui NB. Influence of antioxidants on SOD activity in bovine sperm. Biochem Int 1991; 23(3): 545-53. 8. Bucak MN, Tuncer PB, Sarıözkan S, Başpınar N, Taşpınar M, Çoyan K, Bilgili A, Akalın PP, Büyükleblebici S, Aydos S, Ilgaz S, Sunguroğlu A, Öztuna D. Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: Antioxidants protect DNA integrity against cryodamage. Cryobiology 2010; 61(3): 248-53. 9. Buehr M, Hjorth JP, Hansen AK, Sandoe P. Genetically modified laboratory animals-what welfare problems do they face? J Appl Anim Welf Sci 2003; 6(4): 319-38. 10. Cardullo RA, Baltz JM. Metabolic regulation in mammalian sperm: Mitochondrial volume determines sperm length and flagellar beat frequency. Cell Motil Cytoskeleton 1991; 19(3): 180-8. 11. Charreau B, Tesson L, Soulillou JP, Pourcel C, Anegon I. Transgenesis in rats: Technical aspects and models. Transgenic Res 1996; 5(4): 223-34. cryopreserved mouse ovaries. Hum Reprod 1997; 12(1): 101-6. 12. Cummins JM, Woodall PF. On mammalian sperm dimensions. J Reprod Fertil 1985; 75(1): 153-75. 13. Çaylak E. Hayvan ve bitkilerde oksidatif stres ile antioksidanlar. Tıp Araştırmaları Dergisi 2011; 9(1): 73-83. 14. Çoyan K, Başpınar N, Bucak MN, Akalın PP, Ataman MB, Ömür AD, Güngör Ş, Küçükgünay S, Özkalp B, Sarıözkan S. Influence of methionine and dithioerythritol on sperm motility, lipid peroxidation and antioxidant capacites during liquid storage of ram semen. Res Vet Sci 2010; 89(3): 426-31. 15. Devireddy RV, Swanlund DJ, Roberts KP, Bischof JC. Subzero Water permeability parameters of mouse spermatozoa in the presence of extracellulariIce and cryoprotective agents. Biol Reprod 1999; 61(3): 764-75. 16. Gadea J, Garcia-Vazquez F, Matas C, Gardon JC, Canovas S, Gumbao D. Cooling and freezing of boar spermatozoa: supplementation of the freezing media with reduced glutathione preserves sperm function. J Androl 2005; 26(3): 396-404. 17. Gunasena KT, Villines PM, Critser ES, Critser JK. Live births after autologous transplant of cryopreserved mouse ovaries. Hum Reprod 1997; 12(1): 101-6. 18. Hagiwara M, Choi JH, Devireddy RV, Roberts KP, Wolkers WF, Makhlouf A, Bischof JC. Cellular biophysics during freezing of rat and mouse sperm predicts post-thaw motility. Biol Reprod 2009; 81(4): 700-6. 19. Hall JC, Hadley J, Doman T. Correlation between changes in rat sperm membrane lipid, protein and the membrane physical state during epididymal maturation. J Androl 1991; 12(1): 76-87. 20. Hammerstedt RH, Graham JK, Nolan JP. Cryopreservation of mammalian sperm: What we ask them to survive. J Androl 1990; 11(1): 73-88. 21. Henkel RR. Leukocytes and oxidative stress: Dilemma for sperm function and male fertility. Asian J Androl 2011; 13(1): 43-52. 22. Holt WV. Fundemental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology 2000; 53(1): 47-58. 23. Karunakaran M, Dhali A, Mech A, Khate K, Rajkhowa C, Mishra DP. Preservation of mithun (Bos frontalis) semen at refrigeration temperature. Anim Reprod Sci 2006; 101(3-4): 257-64. 24. Kim JG, Parthasarathy S. Oxidation and the spermatozoa. Semin Reprod Endocrinol 1998; 16(4): 235-9. 25. Landel CP. Archiving mouse strains by cryopreservation. Lab Anim (NY) 2005; 34: 50-7. 26. Lanzafame FM, La Vignera S, Vicari E, Calogero AE. Oxidative stress and medical antioxidant treatment in male infertility. Reprod Biomed Online 2009; 19(5): 638-59. 27. Lazar J, Moreno C, Jacob HJ, Kwitek AE. Impact of genomics on research in the rat. Genome Res 2005; 15: 1717-28. 28. Leibo SP, Songsasen N. Cryopreservation of gametes and embryos of non-domestic species. Theriogenology 2002; 57(1): 303-26. 29. Maxwell WM, Salamon S. Liquid storage of ram semen: A review. Reprod Fertil Dev 1993; 5(6): 613-38. 30. Mazur P, Leibo SP, Seidel GE. Cryopreservation of the germplasm of animals used in biological and medical research: Importance, impact, status and future directions. Biol Reprod 2008; 78(1): 2-12. 31. Nakatsukasa E, Kashiwazaki N, Takizawa A, Shino M, Kitada K, Serikawa T, Hakamata Y, Kobayashi E, Takahashi R, Ueda M, Nakashima T, Nakagata N. Cryopreservation of spermatozoa from closed colonies and inbred spontaneous mutant and transgenic strains of rats. Comp Med 2003; 53(6): 639-41. 32. Parks JE, Lynch DV. Lipid composition and thermotropic behaviour of boar, bull, stallion and rooster sperm membranes. Cryobiology 1992; 29(2): 255-66. 33. Sarıözkan S, Bucak MN, Cantürk F, Özdamar S, Yay A, Tuncer PB, Özcan S, Sorgucu N, Caner Y. The effects of different sugars on motility, morphology and DNA damage during the liquid storage of rat epididymal sperm at 4 0C. Cryobiology 2012; 65(2): 93-7. 34. Sarıözkan S, Özdamar S, Türk G, Cantürk F, Yay A. In vitro effects of L-carnitine and glutamine on motility, acrosomal abnormality, and plasma membrane integrity of rabbit sperm during liquid-storage. Cryobiology 2014; 68(3): 349-53. 35. Sharma RK, Agarwal A. Role of reactive oxygen species in male infertility. Urology 1996; 48(6): 835-50. 36. Si W, Benson JD, Men H, Critser JK. Osmotic tolerance limits and effects of cryoprotectants on the motility, plasma membrane integrity and acrosomal integrity of rat sperm. Cryobiology 2006; 53(3): 336-48. 37. Sonmez M, Turk G, Yuce A. The effect of ascorbic acid supplementation on sperm quality, lipid peroxidation and testosterone levels of male wistar rats. Theriogenology 2005; 63(7): 2063-72. 38. Suarez SS, Marquez B, Harris TP, Schimenti JC. Different regulatory systems operate in the midpiece and principal piece of the mammalian sperm flagellum. Soc Reprod Fertil Suppl 2007; 65: 331-4. 39. Turk G, Ateşşahin A, Sönmez M. Lycopene protects against cyclosporine A-induced testicular toxicity in rats. Theriogenology 2007; 67(4): 778-85. 40. Turk G, Ceribaşı AO, Sakin F, Sönmez M, Ateşşahin A. Antiperoxidative and anti-apoptotoic effects of lycopene and ellagic acid on crclophosphamide-induced testicular lipid peroxidation and apoptosis. Reprod Fertil Dev 2010; 22(4): 587-96. 41. Varışlı O, Uguz C, Agca C, Agca Y. Effect of chilling on the motility and acrosomal integrity of rat sperm in the presence of various extenders. J Am Assoc Lab Anim Sci 2009; 48(5): 499-505. 42. Varışlı O, Uguz C, Agca C, Agca Y. Various physical stres factors on rat sperm motility, integrity of acrosome, and plasma membrane. J Androl 2009; 30(1): 75-86. 43. Varışlı Ö, Ağca C, Ağca Y. Short-term storage of rat sperm in the presence of various extenders. J Am Assoc Lab Anim Sci. 2013; 52(6): 732-7. 44. Verberckmoes S, Van Soom A, Dewulf J, De Pauw I, De Kruif A. Storage of fresh bovine semen in a diluent based on the ionic composition of cauda epididymal plasma. Reprod Dom Anim 2004; 39(6): 410-6. 45. Yamashiro H, Han YJ, Sugawara A, Tomioka I, Hoshino Y, Sato E. Freezability of rat epididymal sperm induced by raffinose in modified Krebs-Ringer bicarbonate (mKRB) based extender solution. Cryobiology 2007; 55(3): 285-94. 46. Yamashrio H, Sato E. Cryopreservation of rat sperm. Katkow I. ed. In: Current Frontiers in Cryopreservation. London: IntechOpen, 2012; pp. 165-78. 47. Yoshida M. Conservation of sperm: Current status and new trends. Anim Reprod Sci 2000; 60(61): 349-55.