Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm

https://doi.org/10.17221/146/2018-VETMEDCitation:Inanc M., Gungor S., Ozturk C., Korkmaz F., Bastan I., Cil B. (2019): Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm. Veterinarni Medicina, 64: 118-124.
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The objective of this study was to determine effects of supplementing Tris-based semen extenders with either cholesterol-loaded cyclodextrin (CLC) or 7-dehydrocholesterol loaded cyclodextrin (7-DCLC) plus trehalose (T) for cryopreservation of ram semen. Semen was collected with an artificial vagina from five Merino rams (2–3 years of age) during the non-breeding season. Ejaculates were pooled, divided into eight equal portions, diluted with a standard Tris-based extender containing: no additive (control); T (50 mM); or T (50 mM) + 1.5, 2.5 or 3.5 mg of either 7-DCLC or CLC. Semen was chilled from 37°C to 4°C, placed in 0.25 ml French straws, held 5 cm above liquid nitrogen for 12 minutes, then plunged into liquid nitrogen. After thawing, a computer-aided semen analyzer system (CASA) was used to assess motility, whereas plasma membrane and acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were assessed with flow cytometry. Sperm supplemented with 2.5 mg and 3.5 mg CLC + T had the highest (P < 0.05) total and progressive motility (65.2 ± 4.7 and 19.0 ± 1.0% respectively, mean ± SEM), albeit with no significant differences from sperm with 1.5 or 3.5 mg CLC + T. Sperm with 2.5 mg CLC + T had the highest (P < 0.05) PMAI (59.3%; not different from 3.5 mg CLC + T) and highest (P < 0.05) HMMP (64.6%; not different from 1.5 or 3.5 mg CLC + T). The lowest ALH value, 2.8 ± 0.3 µm was in the 2.5 mg 7-DCLC + T group; otherwise, there were no significant differences among groups for any other CASA end point. In conclusion, adding CLC + T to a tris-based extender optimized quality of frozen-thawed ram semen. Therefore, extenders including CLC + T have potential to improve quality of frozen-thawed ram sperm.

References:
Aboagla E. M.-E. (2003): Trehalose-Enhanced Fluidity of the Goat Sperm Membrane and Its Protection During Freezing. Biology of Reproduction, 69, 1245-1250 https://doi.org/10.1095/biolreprod.103.017889
 
Ahmad E., Naseer Z., Aksoy M., Küçük N., Uçan U., Serin İ., Ceylan A. (2015): Trehalose enhances osmotic tolerance and suppresses lysophosphatidylcholine-induced acrosome reaction in ram spermatozoon. Andrologia, 47, 786-792 https://doi.org/10.1111/and.12329
 
Aksoy Melih, Akman Orhan, Lehimcioğlu Necdet Cankat, Erdem Hüseyin (2010): Cholesterol-loaded cyclodextrin enhances osmotic tolerance and inhibits the acrosome reaction in rabbit spermatozoa. Animal Reproduction Science, 120, 166-172 https://doi.org/10.1016/j.anireprosci.2010.02.014
 
Blackburn H. D. (2004): Development of national animal genetic resource programs. Reproduction, Fertility and Development, 16, 27- https://doi.org/10.1071/RD03075
 
Blanch E, Tomás C, Graham JK, Mocé E (2012): Response of Boar Sperm to the Treatment with Cholesterol-Loaded Cyclodextrins Added Prior to Cryopreservation. Reproduction in Domestic Animals, 47, 959-964 https://doi.org/10.1111/j.1439-0531.2012.01999.x
 
Bucak Mustafa Numan, Ateşşahin Ahmet, Varışlı Ömer, Yüce Abdurrauf, Tekin Necmettin, Akçay Aytaç (2007): The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen. Theriogenology, 67, 1060-1067 https://doi.org/10.1016/j.theriogenology.2006.12.004
 
Bucak M. N., Ataman M. B., Başpınar N., Uysal O., Taşpınar M., Bilgili A., Öztürk C., Güngör Ş., İnanç M. E., Akal E. (2015): Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity. Andrologia, 47, 545-552 https://doi.org/10.1111/and.12301
 
Cognié Y., Baril G., Poulin N., Mermillod P. (2003): Current status of embryo technologies in sheep and goat. Theriogenology, 59, 171-188 https://doi.org/10.1016/S0093-691X(02)01270-0
 
Cummins James M., Jequier Anne M., Kan Raymond (1994): Molecular biology of human male infertility: Links with aging, mitochondrial genetics, and oxidative stress?. Molecular Reproduction and Development, 37, 345-362 https://doi.org/10.1002/mrd.1080370314
 
De Lamirande E, Gagnon C (1999): Reactive oxygen species and human spermatozoa: I. Effects on the motility of intact spermatozoa and on sperm axonemes. Journal of Andrology 13, 368–378.
 
Garner DL, Hafez ESE (1993): Spermatozoa and seminal plasma. In: Hafez ESE (ed.): Reproduction in Farm Animals. 6th edn. Lea & Febiger, Philadelphia. 165–187.
 
Goularte K.L., Gastal G.D.A., Schiavon R.S., Gonçalves A.O., Schneider J.R., Corcini C.D., Lucia T. (2014): Association between the presence of protein bands in ram seminal plasma and sperm tolerance to freezing. Animal Reproduction Science, 146, 165-169 https://doi.org/10.1016/j.anireprosci.2014.03.009
 
Gungor S., Ozturk C., Omur AD (2017): Positive effects of trehalose and cysteine on ram sperm parameters. Veterinární Medicína, 62, 245-252 https://doi.org/10.17221/131/2016-VETMED
 
Hirano Y, Shibahara H, Obara H, Suzuki T, Takamizawa S, Yamaguchi C, Tsunoda H, Sato I (2001): Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. Journal of Assisted Reproduction and Genetics 18, 215–220.https://doi.org/10.1023/A:1009420432234
 
Iqbal Sajid, Andrabi Syed Murtaza Hassan, Riaz Amjad, Durrani Aneela Zameer, Ahmad Nasim (2016): Trehalose improves semen antioxidant enzymes activity, post-thaw quality, and fertility in Nili Ravi buffaloes ( Bubalus bubalis ). Theriogenology, 85, 954-959 https://doi.org/10.1016/j.theriogenology.2015.11.004
 
Inanc ME, Tekin K, Olgac KT, Ozen D, Stelletta C, Uysal O, Daskin A (2017): Effects of 7-dehydrocholesterol and cholesterol-loaded cyclodextrins on bull sperm motility during short term storage. Journal of the Faculty of Veterinary Medicine Kafkas University 23, 661–664.
 
Inanc ME, Uysal O, Ata A (2018): Cryopreservation and evaluation of Akkaraman ram semen with 7-dehydrocholesterol. Veterinary Journal of Ankara University 65, 187–192.
 
Kasai T, Ogawa K, Mizuno K, Nagai S, Uchida Y, Ohta S, Fujie M, Suzuki K, Hirata S, Hoshi K (2002): Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential. Journal Asian Andrology 4, 97–103.
 
Katanbafzadeh H, Barati F, Tabandeh M (2014): Cryoprotectant-free freezing of the goat epididymal sperm. Cryo Letters 35, 293–298.
 
Khan J, Tahir MZ, Khalid A, Sattar A, Ahmad N (2017): Effect of cholesterol-loaded cyclodextrins on cryosurvival of dog spermatozoa. Reproduction in Domestic Animals, 52, 265-268 https://doi.org/10.1111/rda.12893
 
Konyali C., Tomás C., Blanch E., Gómez E.A., Graham J.K., Mocé E. (2013): Optimizing conditions for treating goat semen with cholesterol-loaded cyclodextrins prior to freezing to improve cryosurvival. Cryobiology, 67, 124-131 https://doi.org/10.1016/j.cryobiol.2013.06.001
 
Korkmaz F., Malama E., Siuda M., Leiding C., Bollwein H. (2017): Effects of sodium pyruvate on viability, synthesis of reactive oxygen species, lipid peroxidation and DNA integrity of cryopreserved bovine sperm. Animal Reproduction Science, 185, 18-27 https://doi.org/10.1016/j.anireprosci.2017.07.017
 
Krause W. (1995): Computer-assisted semen analysis systems: comparison with routine evaluation and prognostic value in male fertility and assisted reproduction. Human Reproduction, 10, 60-66 https://doi.org/10.1093/humrep/10.suppl_1.60
 
López-Revuelta Abel, Sánchez-Gallego José I., Hernández-Hernández Angel, Sánchez-Yagüe Jesús, Llanillo Marcial (2006): Membrane cholesterol contents influence the protective effects of quercetin and rutin in erythrocytes damaged by oxidative stress. Chemico-Biological Interactions, 161, 79-91 https://doi.org/10.1016/j.cbi.2006.03.004
 
Ly JD, Grubb DR, Lawen A (2003): The mitochondrial membrane potential (∆ψm) in apoptosis; an update. Apoptosis 8, 115–128.https://doi.org/10.1023/A:1022945107762
 
Mansour M (2009): Modification of hypo-osmotic swelling test to evaluate the integrity of stallion sperm plasma membrane. Global Veterinaria 3, 302–307.
 
Mocé E., Graham J. K. (2006): Cholesterol-loaded cyclodextrins added to fresh bull ejaculates improve sperm cryosurvival1. Journal of Animal Science, 84, 826-833 https://doi.org/10.2527/2006.844826x
 
Mocé Eva, Purdy Phillip H., Graham James K. (2010): Treating ram sperm with cholesterol-loaded cyclodextrins improves cryosurvival. Animal Reproduction Science, 118, 236-247 https://doi.org/10.1016/j.anireprosci.2009.06.013
 
Moore Amanda I., Squires Edward L., Graham James K. (2005): Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology, 51, 241-249 https://doi.org/10.1016/j.cryobiol.2005.07.004
 
Moraes E.A., Graham J.K., Torres C.A.A., Meyers M., Spizziri B. (2010): Delivering cholesterol or cholestanol to bull sperm membranes improves cryosurvival. Animal Reproduction Science, 118, 148-154 https://doi.org/10.1016/j.anireprosci.2009.08.002
 
Motamedi-Mojdehi R, Roostaei-Ali Mehr M, Rajabi-Toustani R (2014): Effect of Different Levels of Glycerol and Cholesterol-Loaded Cyclodextrin on Cryosurvival of Ram Spermatozoa. Reproduction in Domestic Animals, 49, 65-70 https://doi.org/10.1111/rda.12225
 
Panyaboriban S, Suwimonteerabutr J, Phutikanit N, Swangchan-Uthai T, Tharasanit T, Techakumphu M (2015): Effect of various combinations of sugar supplementation in the extender on frozen-thawed ram semen quality and fertility. Thai Journal of Veterinary Medicine 45, 229–237.
 
Parks John E., Lynch Daniel V. (1992): Lipid composition and thermotropic phase behavior of boar, bull, stallion, and rooster sperm membranes. Cryobiology, 29, 255-266 https://doi.org/10.1016/0011-2240(92)90024-V
 
Pelufo V., López Armengol M.F., Malcotti V., Venturino A., Aisen E.G. (2015): Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm. Theriogenology, 83, 144-151 https://doi.org/10.1016/j.theriogenology.2014.09.007
 
Purdy P.H, Graham J.K (2004): Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm. Cryobiology, 48, 36-45 https://doi.org/10.1016/j.cryobiol.2003.12.001
 
Purdy Phillip H., Mocé Eva, Stobart Robert, Murdoch William J., Moss Gary E., Larson Brent, Ramsey Shawn, Graham James K., Blackburn Harvey D. (2010): The fertility of ram sperm held for 24h at 5°C prior to cryopreservation. Animal Reproduction Science, 118, 231-235 https://doi.org/10.1016/j.anireprosci.2009.06.014
 
Quan GB, Hong QH, Hong QY, Yang HY, Wu SS (2012): The effects of trehalose and sucrose on frozen spermatozoa of Yunnan semi-fine wool sheep during a non-mating season. Cryo Letters 33, 307–317.
 
Spizziri B.E., Fox M.H., Bruemmer J.E., Squires E.L., Graham J.K. (2010): Cholesterol-loaded-cyclodextrins and fertility potential of stallions spermatozoa. Animal Reproduction Science, 118, 255-264 https://doi.org/10.1016/j.anireprosci.2009.08.001
 
Tirpan MB, Tekin N (2015): Effect of boron (sodium pentaborate), added instead of Tris components, on freezing and post-thaw quality of Angora buck semen. Veterinary Journal of Ankara University 62, 295–302.
 
Uçan Uğur, Küçük Niyazi, Ahmad Ejaz, Naseer Zahid, Aksoy Melih, Serin İlker, Ceylan Ahmet (2016): Effect of different sugars supplemented to the extender in combination with cholesterol-loaded cyclodextrin (CLC) on post-thaw quality of ram spermatozoa. Small Ruminant Research, 136, 243-246 https://doi.org/10.1016/j.smallrumres.2016.01.021
 
White IG (1993): Lipids and calcium uptake of sperm in relation to cold shock and preservation: a review. Reproduction, Fertility and Development, 5, 639- https://doi.org/10.1071/RD9930639
 
Zuge RM, Bertolla RP, Nichi M, Soler TBS, Cortada CNM, Bols PEJ, Barnabe VH (2008): Correlation between bovine sperm membrane integrity and mitochondrial activity in Bos taurus bulls. In: 16th International Congress on Animal Reproduction, Budapest, Hungary. July 13.–17. 2008. Abstract 172.
 
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