Effects of ultrasonication on damaged spermatozoa and mitochondrial activity rate

Effects of ultrasonication on damaged spermatozoa and mitochondrial activity rate

The aim of this study was to describe an optimal sonication procedure for sperm cells. Therefore, we used several parameters such as damaged spermatozoa rate (%), mitochondrial activity rate (%), levels of lipid peroxidation, and total antioxidant potential. Ejaculates were collected from rams (n = 3) and were divided into aliquots and 3-, 6-, and 10-s duration times; 1, 3, 5, and 8 repetitive application groups were established. In the groups with 3-, 6- and 10-s duration times, with the increasing number of repeated applications, damaged spermatozoa rates increased (P < 0.05) while mitochondrial activity rates decreased (P < 0.05). In relation with sonication duration time, total antioxidant potential levels increased (P < 0.05) in single-application groups compared to those in control groups and gradually decreased as the repetitions increased. The most effective results were obtained in the group with 8 repetitions and 10-s duration based on damaged spermatozoa rate and mitochondrial activity rate.

___

  • 1. 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 capacities during liquid storage of ram semen. Res Vet Sci 2010; 89: 426–431.
  • 2. Bucak MN, Başpınar N, Tuncer PB, Sarıözkan S, Akalın PP, Çoyan K. Effects of curcumin and dithioerythritol on frozen-thawed bovine semen. Andrologia 2012; 44: 102–109.
  • 3. Nyborg WL. Acoustic Streaming. Vol 2B. New York, NY, USA: Academic Press; 1965.
  • 4. Suslick KS. Ultrasound: Its Chemical, Physical, and Biological Effects. New York, NY, USA: VCH Publishers; 1988.
  • 5. Kuldiloke J. Effect of ultrasound, temperature and pressure treatments on enzyme activity and quality indicators of fruit and vegetable juices. PhD, Technische Universität, Berlin, Germany, 2002.
  • 6. Burden DW. Guide to the disruption of biological samples. Random Primers 2012; 12: 1–25.
  • 7. Halliday D, Resnick R, 1992. Fiziğin Temelleri (Translator: Cengiz Yalçın). 3rd ed. Ankara, Turkey: Arkadaş Press (in Turkish).
  • 8. Thompson LH, Doraiswamy LK. Sonochemistry: science and engineering. Ind Eng Chem Res 1999; 38: 1215–1249.
  • 9. Mercan U. Toksikolojide serbest radikallerin önemi. YYU Vet Fak Derg 2004; 15: 91–96 (in Turkish).
  • 10. Gutteridge JM. Lipid peroxidation and antioxidants as biomarkers of tissue damage. Clin Chem 1995; 4: 1819–1828.
  • 11. Karabiga M, Kiriş İ, Yılmaz N, Altuntaş İ, Karahan N, Okutan H. Aprotinin’in deneysel aortik iskemi reperfüzyon modelinde böbrek hasarı üzerine etkisi. Turkish J Vasc Surg 2007; 165: 9–18.
  • 12. Yajima D, Motani H, Hayakawa M, Sato Y, Sato K, Iwase H. The relationship between cell membrane damage and lipid peroxidation under the condition of hypoxia-reoxygenation: analysis of the mechanism using antioxidants and electron transport inhibitors. Cell Biochem Funct 2009; 27: 338–343.
  • 13. Wong-Ekkabut J, Xu Z, Triampo W, Tang IM, Tieleman DP, Monticelli L. Effect of lipid peroxidation on the properties of lipid bilayers: A molecular dynamics study. Biophys J 2007; 12: 4225–4236.
  • 14. Tateno H, Kimura Y, Yanagimachi R. Sonication per se is not as deleterious to sperm chromosomes as previously inferred. Biol Reprod 2000; 63: 341–346.
  • 15. Yamamoto T, Yoneyama M, Imanishi M, Takeuchi M. Flow cytometric detection and analysis of tailless sperm caused by sonication or a chemical agent. Toxicol Sci 2000; 25: 41–48.
  • 16. Baker SS, Cardullo RA, Thaler CD. Sonication of mouse sperm membranes reveals distinct protein domains. Biol Reprod 2002; 66: 57–64.
  • 17. Evenson DP, Darzynkiewicz Z, Melamed MR. Simultaneous measurement by flow cytometry of sperm cell viability and mitochondrial membrane potential related to cell motility. J Histochem Cytochem 1982; 30: 279–280.
  • 18. Garner DL, Thomas CA, Joerg HW, DeJarnette JM, Marshall CE. Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa. Biol Reprod 1997; 57: 1401–1406.
  • 19. Hallap T, Nagy S, Jaakma U, Johannisson A, Rodriguez-Martinez H. Mitochondrial activity of frozen-thawed spermatozoa assessed by MitoTracker Deep Red 633. Theriogenology 2005; 63: 2311–2322.
  • 20. Edmonds PD, Sancier KM. Evidence for free radical production by ultrasonic cavitation in biological media. Ultrasound Med Biol 1983; 9: 635–639.
  • 21. Milowska K, Gabryelak T, Lypacewicz G, Tymkiewicz R, Nowicki. A. Effect of ultrasound on nucleated erythrocytes. Ultrasound Med Biol 2005; 31: 129–134.
  • 22. Özdemir DS, Başpınar N, Akalın PP. Effects of ultrasound homogenisation on the activities of superoxide dismutase, glutathione peroxidase, catalase and levels of lipid peroxide in liver homogenates. Eurasian J Vet Sci 2015; 31: 16–19.