MUHAMMED ENES İNANÇ, Kemal Tuna OLĞAÇ, Koray TEKİN, Havva ALEMDAR, DOĞUKAN ÖZEN, Ongun UYSAL

Effect of Cholesterol and 7-Dehydrocholesterol on Bull Semen Freezing with Different Rates of Glycerol

The aim of this study was to evaluate the effect of 7-dehydrocholesterol loaded cyclodextrin (7-DHCLC) and cholesterol loaded cyclodextrin (CLC) with the addition of different glycerol (G) rates on bull semen cryopreservation. Semen samples of three bulls were pooled and divided into seven groups; control group (C) and the other six were extended with standard Tris extender including different G rates (3, 6 and 9%) with CLC or 7-DHCLC to reach 1.5 mg/120x106 . Extended samples were frozen in 0.25 mL straws. After thawing, motion characteristics and motility, viability, acrosome integrity, mitochondrial activation, functional membrane integrity and abnormal spermatozoa rates were assessed. The highest membrane integrity, viability and total motility were detected in 7-DHCLC with 9% of G group (P

Kolestrol ve 7-Dehidrokolestrolün Farklı Gliserol Oranları İle Boğa Sperması Dondurulmasına Etkisi Öz

Bu çalışmanın amacı, 7-dehidrokolesterol yüklü siklodekstrin (7-DHCLC) ve kolesterol yüklü siklodekstrin (CLC) ‘in farklı gliserol (G) oranları ilavesi ile boğa sperması dondurulması üzerindeki etkilerini değerlendirmektir. Üç adet boğaya ait sperma örnekleri toplandı, birleştirildi ve yedi gruba ayrıldı; biri kontrol (C) ile, diğer altı grup ise CLC veya 7-DHCLC (1.5 mg/120x106 ) ve farklı G oranlarını (%3, 6 ve 9) içeren standart Tris sulandırıcısı ile sulandırıldı. Sulandırılan spermalar 0.25 mL’lik payetlerde donduruldu. Çözdürme sonrası gruplar hareket karakteristikleri ve motilite, viabilite, akrozom bütünlüğü, mitokondriyal aktivasyon, fonksiyonel membran bütünlüğü ve anormal spermatozoa oranları yönünden değerlendirildi. En yüksek membran bütünlüğü, viabilite ve motilite %9 G ilavesi ile 7-DHCLC grubunda tespit edildi (P

PDF

___

Zeng W, Terada T: Freezability of boar spermatozoa is improved by exposure to 2- hydroxypropyl-beta-cyclodextrin. Reprod Fertil Dev, 12, 223- 228, 2000. DOI: 10.1071/RD00058

Drobnis EZ, Crowe LM, Berger T, Anchordoguy TJ, Overstreet JW, Crowe JH: Cold shock damage is due to lipid phase transitions in cell membranes: A demonstration using sperm as a model. J Exp Zool, 265, 432- 437, 1993. DOI: 10.1002/jez.1402650413

Klein U, Gimpl G, Fahrenholz F: Alteration of the myometrial plasma membrane cholesterol content with b-cyclodextrin modulates the binding affinity of the oxytocin receptor. Biochemistry, 34, 13784-13793, 1995. DOI: 10.1021/bi00042a009

Purdy PH, Graham JK: Effect of adding cholesterol to bull sperm membranes on sperm capacitation, the acrosome reaction, and fertility. Biol Reprod, 71, 522-527, 2004. DOI: 10.1095/biolreprod.103.025577

De Graaf SP, Evans G, Gillan L, Guerra MMP, Maxwell WMC, O’Brien JK: The influence of antioxidant, cholesterol and seminal plasma on the in vitro quality of sorted and non-sorted ram spermatozoa. Theriogenology, 67, 217-227, 2007. DOI: 10.1016/j.theriogenology.2006.07.008

Awad MM: Effects of sub-optimal glycerol concentration and cholesterolloaded cyclodextrin in a tris-based diluent on cryopreserved ram sperm longevity and acrosomal integrity. Small Ruminant Res, 100, 164-168, 2011. DOI: 10.1016/j.smallrumres.2011.05.011

Moce E, Graham JK: Cholesterol- loaded cyclodextrins added to fresh bull ejaculates improve sperm cryosurvival. J Anim Sci, 84, 826-833, 2006.

Tasdemir U, Buyukleblebici S, Tuncer PB, Coskun E, Ozgurtas T, Aydın FN, Buyukleblebici O, Gurcan IS: Effects of various cryoprotectants on bull sperm quality, DNA integrity and oxidative stress parameters. Cryobiology, 66, 38-42, 2013. DOI: 10.1016/j.cryobiol.2012.10.006

Glazer AI, Mullen SF, Liu J, Benson JD, Crister JK, Squines EL, Graham JK: Osmotic tolerance limits and membrane permeability characteristics of stallion spermatozoa treated with cholesterol. Cryobiology, 59, 201-206, 2009. DOI: 10.1016/j.cryobiol.2009.07.009

Gao DY, Ashworth E, Watson PF, Kleinhans FW, Mazur P, Crister JK: Hyperosmotic tolerance of human spermatozoa separate effects of glycerol, sodium chloride, and sucrose on spermolysis. Biol Reprod, 49, 112-123, 1993. DOI: 10.1095/biolreprod49.1.112

Almlid T, Johnson LA: Effects of glycerol concentration, equilibration time and temperature of glycerol addition on post-thaw viability of boar spermatozoa frozen in straws. J Anim Sci, 66, 2899-2905, 1988. DOI: 10.2527/ jas1988.66112899x

Anel L, De Paz P, Alvarez M, Chamorro CA, Boixo JC, Manso A, Gonzalez M, Kaabi M, Anel E: Field and in vitro assay of three methods for freezing ram semen. Theriogenology, 60, 1293-1308, 2003. DOI: 10.1016/ S0093-691X(03)00140-7

Motamedi-Mojdehi R, Roostaei-Ali Mehr M, Rajabi-Toustani R: Effect of different levels of glycerol and cholesterol-loaded cyclodextrin on cryosurvival of ram spermatozoa. Reprod Domest Anim, 49, 65-70, 2014. DOI: 10.1111/rda.12225

Garner DL, Thomas CA, Joerg HW, Dejarnette JM, Marshall CE: Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa. Biol Reprod, 57, 1401-1406, 1997.

Garner DL, Johnson LA: Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol Reprod, 53, 276-284, 1995. DOI: 10.1095/ biolreprod53.2.276

Vanderhost G, Maree L: SpermBlue®: A new universal stain for human and animal sperm which is also amenable to automated sperm morphology analysis. Bioteh Histochem, 84, 299-308, 2009. DOI: 10.3109/ 10520290902984274

Uysal O, Bucak MN: The role of different trehalose concentrations and cooling rates in freezing of ram semen. Ankara Univ Vet Fak Derg, 56, 99-103, 2009.

Tırpan MB, Olğaç KT, Gürler H, Kaya U: Effects of different equilibration conditions on cryopreserved bovine sperm quality. Kocatepe Vet J, 10 (2): 57- 62, 2017.

Fleisch A, Malama, E, Witschi U, Leiding C, Siuda M, Janett F, Bollwein H: Effects of an extension of the equilibration period up to 96 hours on the characteristics of cryopreserved bull semen. Theriogenology, 89, 255-262, 2017. DOI: 10.1016/j.theriogenology.2016.10.018

İnanç ME, Tekin K, Olğaç KT, Özen D, Stelletta C, Uysal O, Daşkın A: Effect of 7-dehydrocholesterol and cholesterol-loaded cyclodextrins on bull sperm motility during short term storage. Kafkas Univ Vet Fak Derg, 23, 661- 664, 2017. DOI: 10.9775/kvfd.2017.17181

Moraes EA, Graham JK, Torres CAA, Meyers M, Spizziri B: Delivering cholesterol or cholestanol to bull sperm membranes improves cryosurvival. Anim Reprod Sci, 118, 148-154, 2010. DOI: 10.1016/j.anireprosci.2009.08.002

Amorim EAM, Graham JK, Spizziri B, Meyers M, Torres CAA: Effect of cholesterol or cholesteryl conjugates on the cryosurvival of bull sperm. Cryobiology, 58, 210-214, 2009. DOI: 10.1016/j.cryobiol.2008.12.007

Blanch E, Tomas C, Hernandez M, Roca J, Martinez EA, Vazquez JM, Moce E: Egg yolk and glycerol requirements for freezing boar spermatozoa treated with methyl-B cyclodextrin or cholesterol-loaded cyclodextrin. J Reprod Dev, 60, 143-149, 2014.

Thomas C, Blanch E, Hernandez M, Gil MA, Roca J, Verquez JM, Martinez EA, Moce E: Treating boar sperm with cholesterol loaded cyclodextrins widens the sperm osmotic tolerance limits and enhances the in vitro sperm fertilising ability. Anim Reprod Sci, 129, 209-220, 2011. DOI: 10.1016/j.anireprosci.2011.12.003

Lopez-Revuelta A, Sanchez-Gallego JL, Hernandez-Hernandez A, Sanchez-Yague Llanillo M: Membrane cholesterol contents influence the protective effects of quercetin and rutin in erythrocytes damaged by oxidative stress. Chem Biol Interact, 161, 79-91, 2006. DOI: 10.1016/j.cbi.2006.03.004

Katanbafzadeh H, Barati F, Tabandeh M: Cryoprotectant-free freezing of the goat epididymal sperm. Cryo Letters, 35, 293-298, 2014.

Mansour MM: Modification of hypo-osmotic swelling test to evaluate the integrity of stallion sperm plasma membrane. Glob Vet, 3, 302-307, 2009.

Moce E, Purdy PH, Graham JK: Treating ram sperm with cholesterolloaded cyclodextrins improves cryosurvival. Anim Reprod Sci, 118, 236-247, 2010. DOI: 10.1016/j.anireprosci.2009.06.013

Moce E, Blanch E, Tomas C, Graham JK: Use of cholesterol in sperm cryopreservation: Present moment and perspectives to future. Reprod Domest Anim, 45, 57-66, 2010. DOI: 10.1111/j.1439-0531.2010.01635.x

Aksoy M, Akman O, Lehimcioglu NC, Erdem H: Cholesterol-loaded cyclodextrin enhances osmotic tolerance and inhibits the acrosome reaction in rabbit spermatozoa. Anim Reprod Sci, 120, 166-172, 2010. DOI: 10.1016/j. anireprosci.2010.02.014

Moore AI, Squires EL, Graham JK: Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology, 51, 241-249, 2005. DOI: 10.1016/j.cryobiol.2005.07.004

Purdy PH, Graham JK: Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm. Cryobiology, 48, 36-45, 2004. DOI: 10.1016/j. cryobiol.2003.12.001

Awad MM, Graham JK: Effect of adding cholesterol to bovine spermatozoa on motility parameters and cell viability after cryopreservation. Cryobiology, 45, 256-257, 2002.

He L, Bailey JL, Buhr MM: Incorporating lipids into boar sperm decreases chilling sensitivity but not capacitation potential. Biol Reprod, 64, 69-79, 2001. DOI: 10.1095/biolreprod64.1.69

De Jonge C: Biological basis for human capacitation-revisited. Hum Reprod Update, 23, 289-299, 2017. DOI: 10.1093/humupd/dmw048

Khorasani AM, Cheung AP, Lee CYG: Cholesterol inhibitory effects on human sperm-induced acrosome reaction. J Androl, 21, 586-594, 2000.

Nolan JP, Graham JK, Hammerstedt RH: Artificial induction of exocytosis in bull sperm. Arch Biochem Biophys, 292, 311-322, 1992. DOI: 10.1016/0003- 9861(92)90084-A

Ehrenwald E, Parks JE, Foote RH: Cholesterol efflux from bovine sperm. I. Induction of the acrosome reaction with lysophosphatidylcholine after reducing sperm cholesterol. Gamete Res, 20, 145-157, 1988. DOI: 10.1002/ mrd.1120200205

Salamon S, Maxwell WMC: Storage of ram semen. Anim Reprod Sci, 62, 77-111, 2000. DOI: 10.1016/S0378-4320(00)00155-X

Storey BT, Noiles EE, Thompson KA: Comparison of glycerol, other polyols, trehalose, and raffinose to provide a defined cryoprotectant medium for mouse sperm cryopreservation. Cryobiology, 37, 46-58, 1998. DOI: 10.1006/cryo.1998.2097

Medeiros CMO, Forell F, Oliveira ATD, Rodrigues JL: Current status of sperm cryopreservation: Why isn’t better. Theriogenology, 57, 327-344, 2002. DOI: 10.1016/S0093-691X(01)00674-4

Hammerstedt RH, Graham JK, Nolan JP: Cryopreservation of mammalian sperm: What we ask them to survive? J Androl, 11, 73-88, 1990.

Mazur P: Freezing of living cells: Mechanisms and implications. Am J Physiol, 247, C125-C142, 1984. DOI: 10.1152/ajpcell.1984.247.3.C125

Guthrie HD, Liu J, Critser JK: Osmotic tolerance limits and effects of cryoprotectants on motility of bovine spermatozoa. Biol Reprod, 67, 1811- 1816, 2002. DOI: 10.1095/biolreprod67.6.1811

Gilmore JA, McGann LE, Liu J, Gao DY, Peter AT, Kleinhans FW, Critser JK: Effect of cryoprotectant solutes on water permeability of human spermatozoa. Biol Reprod, 53, 985-995, 1995. DOI: 10.1095/biolreprod53.5.985