Ayşe DEMİRHAN, Koray TEKİN, Ali DAŞKIN, Ongun UYSAL

Farklı Süre ve Sıcaklıklarda Çözdürülen Boğa Spermalarının Morfolojik Fonksiyonlarının CASA Cihazı ile Değerlendirmesi

Suni tohumlamada kullanılan dondurulmuş spermanın çözüm sonu parametrelerini belirleyen faktörlerin başında çözdürme koşulları gelmektedir. Yapılan araştırma ile Simental, Holstein ve Brown Swiss ırkı boğaların donmuş spermalarının, deneysel olarak belirlenen farklı sıcaklık ve sürelerde çözdürülmesi sonucu CASA cihazı ile spermatozoon başı ve orta kısmına ait parametreler bakımından değerlendirilmesi amaçlanmıştır. Çalışmada gruplar; kontrol grubu 37 ℃’de 20 saniye; deney grupları ise, 25 ℃’de 30, 40, 50 ve 60 sn.; 37 ℃’de 30, 40, 50 ve 60 sn.; 40 ℃’de 10, 15, 20 ve 25 sn.; 70 ℃’de 3, 6, 9, 12 sn. olarak belirlenmiştir. Üç farklı ırk boğaya ait, tek ejakülasyonda elde edilen dondurulmuş boğa spermaları, 9 tekrarda CASA parametreleri açısından değerlendirilmiştir. 25 ℃’de 40 sn.’de çözdürülen spermaların baş uzunluklarının (6.22 ± 0.09), orta kısım genişliğinin (0.68 ± 0.01) diğer sıcaklık ve sürelere göre daha yüksek ortalamaya sahip olduğu saptanmıştır. Çözdürme sıcaklık ve süresinin spermatozoon baş ve orta kısmına dair parametreleri değiştirdiği ancak elde edilen sonuçların istatiksel olarak anlamlı olmadığı belirlenmiştir (p> 0.005).

Anahtar Kelimeler:

Boğa spermatozoonu, CASA, morfoloji, spermatozoon başı, spermatozoon orta kısmı

Assessment of Morphological Features of Bull Semen Thawed at Various Temperatures and Periods of Time with CASA

Thawing condition is one the most important factors affecting the re-animation of the spermatozoon in order to fertilise the oocyte. For that matter, we aimed to evaluate the morphological features of the head and midpiece of cryopreserved spermatozoa thawed at different temperatures and for various durations, with using CASA (Computer aided sperm analyser). Frozen semen samples belonging to the same batch, collected from three different bulls were grouped as; control group thawed for 20 seconds at 37 ℃; and experimental groups were thawed for 30, 40, 50 and 60 s at 25 ℃ and 37 ℃; for 10, 15, 20 and 25 s at 40 ℃; for 3, 6, 9, 12 s at 70 ℃. Morphometrical features of the samples were evaluated by using CASA system with nine repetitions. As a result, straws thawed at 25℃ for 40 s had the highest average length of head (6.22 ± 0.09 μm), and the width of mid-piece (0.68 ± 0.01 μm). It was concluded that the thawing temperature and duration has affected/altered the morphometry of the sperm head and midpiece, although the results were not statistically significant (p> 0.005).

Keywords:

Bull sperm, CASA, morphometry, sperm head, sperm mid-piece,

PDF

___

Aitken RJ. Free radicals, lipid peroxidation and sperm function. Reprod. Fertil. Dev. 1995; 7: 659–68.
Alvarez JG, Touchstone JC, Blasco L, Storey BT. Spontaneous lipid per-oxidation and production of hydrogen peroxide and superoxide in humanspermatozoa. Superoxide dismutase as major enzyme protectant againstoxygen toxicity. J. Androl. 1987; 8: 338–48.
Alvarez JG, Storey BT. Differential incorporation of fatty acids into andperoxidative loss of fatty acids from phospholipids of human spermatozoa. Mol. Reprod. Dev. 1995; 42: 334–46.
Anderson MJ ve Dixson AF. Motility and the mid-piece in primates. Nature. 2002; 416: 496.
Beletti ME, Costa Lda F, Viana MP. A comparison of morphometric characteristics of sperm from fertile Bos taurus and Bos indicus bulls in Brazil. Anim Reprod Sci. 2005; 85: 105–16.
Bennison C, Hemmings N, Slate J, Birkhead T. Long sperm fertilize more eggs in a bird. Proc Biol Sci. 2015; 282(1799): 20141897.
Buhr MM, Canvin AT, Bailey JL. Effects of semen preservation on boarspermatozoa head membranes. Gamete Res. 1989; 23: 441–449.
Calamera JC, Brugo S, Vilar O. Relation between motility and adenosine tri phosphate (ATP) in human spermatozoa. Andrologia.1982; 14: 239–41.
Calamera JC, Buffone MG, Doncel GF, Brugo Olmeda S, Vincentiis S, Calamera MM, Storey BT, Alvarez JG. Effect of thawing temperature on the motility recovery of cryopreserved human spermatozoa. Fertility and Sterility. 2010; 93(3): 789-794.
Cardullo RA ve Baltz JM. Metabolic regulation inmammalian sperm: mitochondrial volume determines sperm length and flagellar beat frequency. Cell Motil. Cytoskeleton. 1991; 19: 180–188.
Chatterjee S, Gagnon C. Production of reactive oxygen species by sper-matozoa undergoing cooling, freezing, and thawing. Mol. Reprod. Dev. 2001; 59: 451–458.
Cummins JM ve Woodall PF. On mammalian sperm dimensions. J. Reprod. Fertil. 1985; 75: 153–175.
Dadoune PJ, Mayaux MJ, Guihard-Moscato ML. Correlation between defects in chromatin condensation of human spermatozoa stained by aniline blue and semen characteristics. Andrologia. 1988; 20: 211–217.
Desai N, Sabanegh E, Kim T, Agarwal A. Free radical theory of aging: Implication in male fertility. Urology. 2010; 75: 14-19.
Firman RC, Simmons LW. Sperm midpiece length predicts sperm swimmingvelocity in house mice. Biol Lett. 2010; 6(4): 513–6.
Fisher HS, Palmer EJ, Lassance JM, Hoekstra HE. The genetic basis and fitness consequences of sperm midpiece size in deer mice. Nat Commun. 2016; 7: 13652.
Fitzpatrick JL, Montgomerie R, Desjardins JK, Stiver KA, Kolm N ve Balshine S. Female promiscuitypromotes the evolution of faster sperm in cichlid fishes. Proc. Natl Acad. Sci. USA. 2009; 106: 1128–1132.
García‑Herreros M, Barón FJ, Aparicio IM, Santos AJ, Gacría‑Marín LJ. Morphometric changes in boar spermatozoa induced by cryopreservation. Int J Androl. 2007; 31: 490–498.
Glander HJ, Schaller J. Binding of annexin V to plasma membranes ofhuman spermatozoa: a rapid assay for detection of membrane changesafter cryostorage. Mol. Hum. Reprod. 1999; 5: 109–15.
Gomendio M ve Roldan ERS. Sperm size and sperm competition in mammals. Proc. Roy. Soc. London B. 1991; 243: 181-185.
Gravance CG, Vishwanath R, Pitt C, Garner DL, Casey PJ. Effects of cryopreservation on bull sperm head morphometry. J. Androl.1998; 19: 704–709.
Gravance CG, Casey ME, Casey PJ. Pre‑freeze bull sperm head morphometry related to post‑thaw fertility. Anim Reprod Sci, 2009; 114: 81–88.
Gil, M. C., García-Herreros, M., Barón, F. J., Aparicio, I. M., Santos, A. J., & García-Marín, L. J. (2009). Morphometry of porcine spermatozoa and its functional significance in relation with the motility parameters in fresh semen. Theriogenology, 71(2), 254-263.
Gu NH, Zhao WL, Wang GS, Sun F. Comparative analysis of mammalian sperm ultrastructure reveals relationships between sperm morphology, mitochondrial functions and motility. Reproductive Biology and Endocrinology. 2019; 17: 1-12.
Hingst O, Blottner S, Franz C. Chromatin condensation in cat spermatozoa during epididymal transit as studied by aniline blue and acridine orange staining. Andrologia. 1995; 27: 275–279.
İleri IK and AK K. Payet yontemine gore dondurulmus boga spermas1n1n eritilmesinde eritme ısısı ve surelerinin spermaztozoitlerin motilite ve akrozom yapıları uzerine etkileri. Istanbul Universitesi Veteriner Fakultesi ve Munich Ludwing-Maximilian Universitesi Veteriner Fakultesi Turk-Alman Gunleri 1993 ; 29-30 Nisan-Mayıs Tebligler, 58-62.
Januskauskas A, Johannison A, Rodriguez-martinez H. Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure and ﬁeld fertility. Theriogenology. 2003; 60: 743–758.
Jones R, Mann T, Sherins R. Peroxidative breakdown of phospholipids inhuman spermatozoa, spermicidal properties of fatty acid peroxides, andprotective action of seminal plasma. Fertil. Steril. 1979; 31: 531–537.
Karabinus DS, Evenson DP, Kaproth MT. Effects of egg yolk-citrate and milk extenders on chromatin structure and viability of cryopreserved bull sperm. J. Dairy Sci. 1991; 74: 3836–3848.
Karabinus DK, Vogler CJ, Saacke RG, Evenson DP. Chromatin structural changes in sperm after scrotal insulation in Holstein bulls. J Androl. 1997; 18: 549–555.
Lüpold S, Calhim S, Immler S, Birkhead TR. Sperm morphology and sperm velocity in passerinebirds. Proc. R. Soc. B. 2009; 276: 1175–1181.
Malo AF, Gomendio M, Garde J, Lang-Lenton B, Soler AJ, Roldan ERS. Sperm design andsperm function. Biol. Lett. 2006; 2: 246–249.
Maroto-Moralesa A, Ramona M, Garcıa-Alvarezb O, Solera AJ, Estesoc MC, Martinez-Pastord F, Perez-Guzmanb MD, Gardea JJ. Characterization of ram (Ovis aries) sperm head morphometryusing the Sperm-Class Analyzer. Theriogenolgy. 2010; 73: 437-473.
Núñez‑Martínez I, Moran JM, Peña FJ. Sperm indexes obtained using computer‑assisted morphometry provide a forecast of the freezability of canine sperm. Int J Androl. 2007; 30: 182–9.
Nur Z, Doğan I, Soylu MK, Ak K. Effect of different thawing procedures on the quality of bull semen. Revue Méd. Vét. 2003; 154(7): 487-490.
Mortimer D. The functional anatomy of the human spermatozoon: relatingultrastructure and function. Mol. Hum. Reprod. 2018; 24(12): 567–92.
Ostermeier GC, Sargeant GA, Yandell BS, Evenson DP, Parrish JJ. Relationship of Bull Fertility to Sperm Nuclear Shape. Journal of Andrology. 2001; 22(4): 595-603.
Piomboni P, Focarelli R, Stendardi A, Ferramosca A, Zara V. The role of mitochondria in energy production forhuman sperm motility. Int J Androl. 2012; 35(2): 109–24.
Ramm SA, Parker GA, Stockley P. Sperm competition and the evolution ofmale reproductive anatomy in rodents. Proc Biol Sci. 2005; 272(1566): 949–55.
Royere D, Hamamah S, Nicolle C, Barthelemy C, Lansac J. Freezing and thawing alter chromatin stability of ejaculated human spermatozoa: ﬂuorescence acridine orange staining and Fuelgen-DNA cytophotometric studies. Gamete Res. 1988; 21: 51–57.
Saacke RG, White JM. Semen quality tests and their relationship to fertility. In: NAAB Proceedings of the Fourth Technical Conference on Artiﬁcial Insemination and Reproduction. 1972; p: 22–27.
Sailer BL, Jost LK, Evenson DP. Mammalian sperm DNA susceptibilityto in situ denaturation associated with the presence of DNA strand breaksas measured by the terminal deoxynucleotidyl transferase assay. J. Androl. 1995; 16: 80–87.
Sailer BL, Jost LK, Evenson DP. Bull sperm head morphometry related to abnormal chromatin structure and fertility. Cytometry. 1996; 24: 167–173.
Schiller J, Arnhold J, Glander HJ, Arnold K. Lipid analysis of human spermatozoa and seminal plasma by MALDI-TOF mass spectrometryand NMR spectroscopy—effects of freezing and thawing. Chem PhysLipids. 2000; 106: 145–156.
Senger PL. : Handling frozen bovine semen-factors which influence viability and fertility. Theriogenology. 1980; 13: 51-62.
Shahani SK, Revell SG, Argo CG, Murray RD. Mid-piece Length of Spermatozoa in Different Cattle Breeds and its Relationship to Fertility. Pakistan Journal of Biological Sciences. 2010; 13(2): 802-808.
Shivaji S, Kota V, Siva AB. The role of mitochondrial proteins in sperm capacitation. J. Reproduct. Immunol. 2009; 83: 14-18.
Yániz J, Soler C, Santolaria P. Computer assisted sperm morphometry in mammals: a review. Anim Reprod Sci. 2015; 156:1–12.
Valverde A, Arenán H, Sancho M, Contell J, Yániz J, Fernández A, Soler C. Morphometry and subpopulation structure of Holstein bull spermatozoa: variations in ejaculates and cryopreservation straws. Asian Journal of Andrology. 2016; 18: 851-857.