اثر سانتریفیوژ و کلسترول بارگذاری‌شده با سیکلودکسترین در رقیق‌کننده لیسیتین سویا بر‌کیفیت اسپرم قوچ قزل بعد از فرآیند ذوب
محورهای موضوعی :
آسیب شناسی درمانگاهی دامپزشکی
محمد شمس الهی
1
(دانشجوی دکترای تخصصی گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.)
حسین دقیق کیا
2
(استاد گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.)
تاریخ دریافت : 1397/03/11
تاریخ پذیرش : 1398/02/11
تاریخ انتشار : 1398/09/01
کلید واژه:
اسپرم,
قوچ,
سانتریفیوژ,
سیکلودکسترین,
انجماد-ذوب,
چکیده مقاله :
حفظ تحرک و زنده مانی اسپرم قوچ هنگام استفاده از اسپرم منجمد شده برای به دست آوردن میزان آبستنی بالا مهم است. هدف از این مطالعه بررسی اثر سانتریفیوژ و کلسترول بارگذاری شده با سیکلودکسترین در لیسیتین سویا به عنوان رقیق کننده، بر کیفیت اسپرم بعد از فرایند انجماد-ذوب بود. نمونه‌های منی از 5 رأس قوچ قزل 5-4 ساله به مدت 3 هفته جمع‌آوری شدند. بعد از ارزیابی اولیه و تائید کیفیت، نمونه‌ها به 8 قسمت مساوی تقسیم شدند. پلاسمای منی 4 نمونه در دمای 30 درجه سلسیوس به وسیله سانتریفیوژ حذف گردید. نمونه‌های باقی مانده بدون سانتریفیوژ به وسیله بافر تریس و غلظت‌های مختلف کلسترول لودشده با سیکلودکسترین (0، 75/0، 5/1 و 3 میلی‌گرم در 108×2/1 اسپرم) و 7 درصد گلیسرول مشابه با گروه اول رقیق‌سازی شدند. سپس نمونه‌ها تا دمای 5 درجه سلسیوس سردسازی شده و در پایوت‌های 25/0 میلی‌لیتری ذخیره شدند. نتایج نشان‌دهنده بهبود معنی‌دار پارامترهای تحرک کل و پیش‌رونده، سرعت در مسیر مستقیم، سرعت در مسیر منحنی، تناوب عرضی زنش، یکپارچگی غشای آکروزومی، یکپارچگی غشای‌پلاسمایی، ظرفیت تام آنتی اکسیدانی و فعالیت آنزیم های سوپراکسید دیسموتاز، گلوتاتیون پراکسیداز و مالون دی‌آلدئید، بعد از ذوب در گروه دریافت کننده 5/1 میلی‌گرم سیکلودکسترین در مقایسه با سایر گروه ها بود (05/0p <). همچنین تحرک کل و پیش‌رونده، سرعت در مسیر منحنی، یکپارچگی غشای پلاسمایی، ظرفیت تام آنتی‌اکسیدانی و فعالیت آنزیم های سوپراکسید دیسموتاز و گلوتاتیون پراکسیداز به طور معنی‌داری در گروه بدون سانتریفیوژ نسبت به گروه با سانتریفیوژ، بالاتر بود (05/0p <). به طورکلی نتایج نشان داد که افزودن 5/1 میلی‌گرم کلسترول بارگذاری شده با سیکلودکسترین باعث بهبود اغلب پارامترهای مورد ارزیابی نسبت به سایر گروه‌ها شد.
چکیده انگلیسی:
When using frozen-thawed semen, maintaining the motility and viability of ram sperm is important for achieving high pregnancy rates. The goal of this research was to clearly elucidate the effects of centrifugation and cholesterol-loaded cyclodextrin (CLC) in a soy lecithin-based extender on post-thaw ram sperm quality. Sperm samples were collected from 5 Ghezel rams with the age of 4-5 years for 3 weeks. After the initial assessment, the approved semen samples were combined and divided into 8 equal parts. Four samples were combined at 30˚C and the seminal plasma was then removed by centrifugation. The semen from the other four samples was not centrifuged and was diluted with Tris buffer plus different concentrations of CLC (0, 0.75, 1.5 and 3 mg/ 1.2×108 spermatozoa) and 7% glycerol, similar to the first group. The samples were then cooled to 5˚C and frozen in 0.25 ml straws. After thawing, total motility (TM) and progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL), beat cross frequency (BCF), total antioxidant capacity (TAC), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPX), the integrity of the acrosome membrane and the integrity of the plasma membrane were significantly (p < /em><0.05) higher in the 1.5 mg CLC group compared to the other groups. Also TM, PM, VCL, TAC, SOD, GPX and plasma membrane was significantly (p < /em><0.05) higher in the group without centrifugation than in the centrifugation group. Overall, the results suggested that addition of 1.5 mg CLC improves most of the sperm parameters measured in vitro in comparison to other groups.
منابع و مأخذ:
Ahmad, E., Aksoy, M., Serin, I., Kucuk, N., Ceylan, A., Ucan, U., et al. (2013). Cholesterol-loaded cyclodextrin pretreatment of ram spermatozoa protects structural integrity of plasma membrane during osmotic challenge and reduces their ability to undergo acrosome reaction in vitro. Small Ruminant Research, 115(1-3): 77-81.
Awad, M.M. (2011). Effects of sub-optimal glycerol concentration and cholesterol-loaded cyclodextrin in a Tris-based diluent on cryopreserved ram sperm longevity and acrosomal integrity. Small Ruminant Research, 100(2-3): 164-168.
Bilodeau, J.F., Chatterjee, S., Sirard, M.A. and Gagnon, C. (2000). Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Molecullar Reproduction and Development, 55(3): 282-288.
Bucak, M.N., Atessahin, A., Varisli, Ö., Yüce, A., Tekin, N., Akcay, A., et al. (2007). Influence of trehalose, taurine, cysteamine and hyaluronan on ram semen microscopic and oxidative stress parameters after freeze thawing process. Theriogenology, 67(5): 1060-1067.
Combes, G.B., Varner, F., Schroeder, R.C. and Burghardt, T.L. (2000). Effect of cholesterol on the motility and plasma membrane integrity of frozen equine spermatozoa after thawing. Journal of reproduction and fertility. Supplement, 56(56): 127-132.
Cross, N.L. (1998). Role of cholesterol in sperm capacitation, Biology Reproduction, 59(1): 7-11.
Cross, N.L. (2003). Decrease in order of human sperm lipids during capacitation. Biology of Reproduction, 69(2): 529-534.
Darin-Bennett, A. and White, I.G. (1977). Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock. Cryobiology, 14(4): 466-470.
Del, E., Olmo, A., Bisbal, A., Maroto-Morales, O., Garcia-Alvarez, M., Ramon, P., et al. (2013). Fertility of cryopreserved ovine semen is determined by sperm velocity. Animal Reproduction Science, 138(1-2): 102-109.
Elizabeth, G., Crichton, A., Budhan, S., Pukazhenthi, B., Billah, A., Julian, A., et al. (2015). Cholesterol addition aids the cryopreservation of dromedary camel (Camelus dromedarius) spermatozoa. Theriogenology, 83(2): 168-174.
Ellington, J.E., Samper, J.C., Jones, A.E., Oliver, S.A., Burnett, K.M., Wright, R.W., et al. (1999). In vitro interactions of cryopreserved stallion spermatozoa and oviduct (uterine tube) epithelial cells or their secretory products. Animal Reproduction Science, 56(1): 51-65.
Esterbauer, H. and Cheeseman, K.H. (1990). Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods in Enzymology, 186 407-421.
Futino, D., Mendes, M., Matos, W., Mondadori, R. and Lucci, C. (2010). Glycerol, methyl-formamide and dimethyl-formamide in canine semen cryopreservation. Reproduction in Domestic Animals, 45(2): 214-220.
Gil, J., Söderquist, L. and Rodríguez-Martínez, H. (1999). Influence of centrifugation and different extenders on post-thaw sperm quality of ram sperm. Theriogenology, 54(1): 93-108.
Hancock, J.L. (1956). The morphology of boar spermatozoa. Journal of the Royal Microscopical Society, 76(3): 84-97.
Holt, W.V. (2000). Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology, 53(1): 47-58.
Miller, J.K., Brzezinska-Slebodzinska, E. and Madsen, F.C. (1993). Oxidative stress, antioxidants, and animal function. Journal of Dairy Science, 76(9): 2812-2823.
Moore, A.I., Squires, E.L. and Graham, J.K. (2005). Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology, 51(3): 241-249.
Naseer, Z., Ahmad, E., Aksoy, M., Küçük, N., Serin, İ., Ceylan, A., et al. (2015). Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm. Cryobiology, 71(1): 18-23.
Ochsendorf, F. (1999). Infections in the male genital tract and reactive oxygen species. Human Reproduction Update, 5(5): 399-420.
Ohvo-Rekila, H., Ramstedt, B., Leppimaki, P. and Slotte J.P. (2002). Cholesterol interactions with phospholipids in membranes. Progress Lipid Research, 41(1): 66-97.
Partyka, A., Łukaszewicz, E. and Niżański, W. (2012a). Effect of cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activity in fowl semen. Theriogenology, 77(8): 1497-1504.
Partyka, A., Łukaszewicz, E. and Niżański, W. (2012b). Lipid peroxidation and antioxidant enzymes activity in avian semen. Animal Reproduction Science, 134(3-4): 184-190.
Purdy, P.H. and Graham J.K. (2004). Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm. Cryobiology, 48(1): 36-45.
Ritar, A.J. (1993). Control of ovulation, storage of semen and artificial insemination of fiber-producing goats in Australia a review. Australian Journal of Experimental Agriculture, 33: 807-820.
Roostaei, A.M.M., Mousavi, M. and Ghadamyari, M. (2015). Effect of seminal plasma proteins on membrane cholesterol efflux of ram epididymal spermatozoa. Small Ruminant Research, 129: 88-91.
Salmani, H., Nabi, M.M., Vaseghi-Dodaran, H., Rahman, M.B., Mohammadi-Sangcheshmeh, A., Shakeri, M., et al. (2013). Effect of glutathione in soybean lecithin-based semen extender on goat semen quality after freeze-thawing. Small Ruminant Research, 112(1-3): 123-127.
Serin, I., Aksoy, M. and Ceylan, A. (2011). Cholesterol-loaded cyclodextrin inhibits premature acrosomal reactions in liquid-stored rabbit spermatozoa. Animal Reproduction Science, 123(1-2): 106-111.
Sion, B., Janny, L., Boucher, D. and Grizard, G. (2004). Annexin V binding to plasma membrane predicts the quality of human cryopreserved spermatozoa. International Journal of Andrology, 27(2): 108-114.
Thys, M., Nauwynck, H., Maes, D., Hoogewijs, M., Vercauteren, D., Rijsselaere, T., Favoreel, H., Van Soom, A., et al. (2009). Expression and putative function of fibronectin and its receptor (integrin a5b1) in male and female gametes during bovine fertilization in vitro. Reproduction 138(3): 471-482.
Upreti, G.C., Hall, E.L., Koppens, D., Oliver, J.E. and Vishwanath, R. (1999). Studies on the measurement of phospholipase A2 (PLA2) and PLA2 inhibitor activities in ram semen. Animal Reproduction Science, 56(2): 107-121.
Witte, T.S. and Schäfer-Somi, S. (2007). Involvement of cholesterol, calcium and progesterone in the induction of capacitation and acrosome reaction of mammalian spermatozoa. Animal and Reproduction Science, 102(3-4): 181-193.
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Ahmad, E., Aksoy, M., Serin, I., Kucuk, N., Ceylan, A., Ucan, U., et al. (2013). Cholesterol-loaded cyclodextrin pretreatment of ram spermatozoa protects structural integrity of plasma membrane during osmotic challenge and reduces their ability to undergo acrosome reaction in vitro. Small Ruminant Research, 115(1-3): 77-81.
Awad, M.M. (2011). Effects of sub-optimal glycerol concentration and cholesterol-loaded cyclodextrin in a Tris-based diluent on cryopreserved ram sperm longevity and acrosomal integrity. Small Ruminant Research, 100(2-3): 164-168.
Bilodeau, J.F., Chatterjee, S., Sirard, M.A. and Gagnon, C. (2000). Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Molecullar Reproduction and Development, 55(3): 282-288.
Bucak, M.N., Atessahin, A., Varisli, Ö., Yüce, A., Tekin, N., Akcay, A., et al. (2007). Influence of trehalose, taurine, cysteamine and hyaluronan on ram semen microscopic and oxidative stress parameters after freeze thawing process. Theriogenology, 67(5): 1060-1067.
Combes, G.B., Varner, F., Schroeder, R.C. and Burghardt, T.L. (2000). Effect of cholesterol on the motility and plasma membrane integrity of frozen equine spermatozoa after thawing. Journal of reproduction and fertility. Supplement, 56(56): 127-132.
Cross, N.L. (1998). Role of cholesterol in sperm capacitation, Biology Reproduction, 59(1): 7-11.
Cross, N.L. (2003). Decrease in order of human sperm lipids during capacitation. Biology of Reproduction, 69(2): 529-534.
Darin-Bennett, A. and White, I.G. (1977). Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock. Cryobiology, 14(4): 466-470.
Del, E., Olmo, A., Bisbal, A., Maroto-Morales, O., Garcia-Alvarez, M., Ramon, P., et al. (2013). Fertility of cryopreserved ovine semen is determined by sperm velocity. Animal Reproduction Science, 138(1-2): 102-109.
Elizabeth, G., Crichton, A., Budhan, S., Pukazhenthi, B., Billah, A., Julian, A., et al. (2015). Cholesterol addition aids the cryopreservation of dromedary camel (Camelus dromedarius) spermatozoa. Theriogenology, 83(2): 168-174.
Ellington, J.E., Samper, J.C., Jones, A.E., Oliver, S.A., Burnett, K.M., Wright, R.W., et al. (1999). In vitro interactions of cryopreserved stallion spermatozoa and oviduct (uterine tube) epithelial cells or their secretory products. Animal Reproduction Science, 56(1): 51-65.
Esterbauer, H. and Cheeseman, K.H. (1990). Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods in Enzymology, 186 407-421.
Futino, D., Mendes, M., Matos, W., Mondadori, R. and Lucci, C. (2010). Glycerol, methyl-formamide and dimethyl-formamide in canine semen cryopreservation. Reproduction in Domestic Animals, 45(2): 214-220.
Gil, J., Söderquist, L. and Rodríguez-Martínez, H. (1999). Influence of centrifugation and different extenders on post-thaw sperm quality of ram sperm. Theriogenology, 54(1): 93-108.
Hancock, J.L. (1956). The morphology of boar spermatozoa. Journal of the Royal Microscopical Society, 76(3): 84-97.
Holt, W.V. (2000). Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology, 53(1): 47-58.
Miller, J.K., Brzezinska-Slebodzinska, E. and Madsen, F.C. (1993). Oxidative stress, antioxidants, and animal function. Journal of Dairy Science, 76(9): 2812-2823.
Moore, A.I., Squires, E.L. and Graham, J.K. (2005). Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology, 51(3): 241-249.
Naseer, Z., Ahmad, E., Aksoy, M., Küçük, N., Serin, İ., Ceylan, A., et al. (2015). Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm. Cryobiology, 71(1): 18-23.
Ochsendorf, F. (1999). Infections in the male genital tract and reactive oxygen species. Human Reproduction Update, 5(5): 399-420.
Ohvo-Rekila, H., Ramstedt, B., Leppimaki, P. and Slotte J.P. (2002). Cholesterol interactions with phospholipids in membranes. Progress Lipid Research, 41(1): 66-97.
Partyka, A., Łukaszewicz, E. and Niżański, W. (2012a). Effect of cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activity in fowl semen. Theriogenology, 77(8): 1497-1504.
Partyka, A., Łukaszewicz, E. and Niżański, W. (2012b). Lipid peroxidation and antioxidant enzymes activity in avian semen. Animal Reproduction Science, 134(3-4): 184-190.
Purdy, P.H. and Graham J.K. (2004). Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm. Cryobiology, 48(1): 36-45.
Ritar, A.J. (1993). Control of ovulation, storage of semen and artificial insemination of fiber-producing goats in Australia a review. Australian Journal of Experimental Agriculture, 33: 807-820.
Roostaei, A.M.M., Mousavi, M. and Ghadamyari, M. (2015). Effect of seminal plasma proteins on membrane cholesterol efflux of ram epididymal spermatozoa. Small Ruminant Research, 129: 88-91.
Salmani, H., Nabi, M.M., Vaseghi-Dodaran, H., Rahman, M.B., Mohammadi-Sangcheshmeh, A., Shakeri, M., et al. (2013). Effect of glutathione in soybean lecithin-based semen extender on goat semen quality after freeze-thawing. Small Ruminant Research, 112(1-3): 123-127.
Serin, I., Aksoy, M. and Ceylan, A. (2011). Cholesterol-loaded cyclodextrin inhibits premature acrosomal reactions in liquid-stored rabbit spermatozoa. Animal Reproduction Science, 123(1-2): 106-111.
Sion, B., Janny, L., Boucher, D. and Grizard, G. (2004). Annexin V binding to plasma membrane predicts the quality of human cryopreserved spermatozoa. International Journal of Andrology, 27(2): 108-114.
Thys, M., Nauwynck, H., Maes, D., Hoogewijs, M., Vercauteren, D., Rijsselaere, T., Favoreel, H., Van Soom, A., et al. (2009). Expression and putative function of fibronectin and its receptor (integrin a5b1) in male and female gametes during bovine fertilization in vitro. Reproduction 138(3): 471-482.
Upreti, G.C., Hall, E.L., Koppens, D., Oliver, J.E. and Vishwanath, R. (1999). Studies on the measurement of phospholipase A2 (PLA2) and PLA2 inhibitor activities in ram semen. Animal Reproduction Science, 56(2): 107-121.
Witte, T.S. and Schäfer-Somi, S. (2007). Involvement of cholesterol, calcium and progesterone in the induction of capacitation and acrosome reaction of mammalian spermatozoa. Animal and Reproduction Science, 102(3-4): 181-193.
