- Volume 29 Issue 5
DOI QR Code
Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa
- Prapaiwan, N. (Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University) ;
- Tharasanit, T. (Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University) ;
- Punjachaipornpol, S. (Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University) ;
- Yamtang, D. (Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University) ;
- Roongsitthichai, A. (Department of Veterinary Clinical Science, Faculty of Veterinary Sciences, Mahasarakham University) ;
- Moonarmart, W. (Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University) ;
- Kaeoket, K. (Department of Clinical Sciences and Public Health Medicine, Faculty of Veterinary Science, Mahidol University) ;
- Manee-in, S. (Department of Clinical Sciences and Public Health Medicine, Faculty of Veterinary Science, Mahidol University)
- Received : 2015.07.06
- Accepted : 2015.09.12
- Published : 2016.05.01
Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL) has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05) than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05). In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.
Low-density Lipoprotein;Epididymal Spermatozoa;Cryopreservation;Dog
Supported by : Mahidol University
- Aitken, R. J. and E. A. McLaughlin. 2007. Molecular mechanisms of sperm capacitation: Progesterone-induced secondary calcium oscillations reflect the attainment of a capacitated state. Soc. Reprod. Fertil. Suppl. 63:273-293.
- Amirat, L., D. Tainturier, L. Jeanneau, C. Thorin, O. Gerard, J. L. Courtens, and M. Anton. 2004. Bull semen in vitro fertility after cryopreservation using egg yolk LDL: A comparison with Optidyl, a commercial egg yolk extender. Theriogenology 61:895-907. https://doi.org/10.1016/S0093-691X(03)00259-0
- Andersen, K. 1975. Insemination with frozen dog semen based on a new insemination technique. Reprod. Domest. Anim. 10:1-4. https://doi.org/10.1111/j.1439-0531.1975.tb00275.x
- Anton, M., V. Martinet, M. Dalgalarrondo, V. Beaumal, E. David-Briand, and H. Rabesona. 2003. Chemical and structural characterisation of low-density lipoproteins purified from hen egg yolk. Food Chem. 83:175-183. https://doi.org/10.1016/S0308-8146(03)00060-8
- Axner, E., U. Hermansson, and C. Linde-Forsberg. 2004. The effect of Equex STM paste and sperm morphology on post-thaw survival of cat epididymal spermatozoa. Anim. Reprod. Sci. 84:179-191. https://doi.org/10.1016/j.anireprosci.2003.11.003
- Bedford, J. M. 1968. Ultrastructural changes in the sperm head during fertilization in the rabbit. Am. J. Anat. 123:329-358. https://doi.org/10.1002/aja.1001230207
- Bencharif, D., L. Amirat-Briand, A. Garand, M. Anton, E. Schmitt, S. Desherces, G. Delhomme, M. L. Langlois, P. Barriere, S. Destrumelle, O. Vera-Munoz, and D. Tainturier. 2010. Freezing canine sperm: Comparison of semen extenders containing Equex and LDL (low density lipoproteins). Anim. Reprod. Sci. 119:305-313. https://doi.org/10.1016/j.anireprosci.2010.01.009
- Bencharif, D., L. Amirat, M. Anton, E. Schmitt, S. Desherces, G. Delhomme, M. L. Langlois, P. Barriere, M. Larrat, and D. Tainturier. 2008. The advantages of LDL (low density lipoproteins) in the cryopreservation of canine semen. Theriogenology 70:1478-1488. https://doi.org/10.1016/j.theriogenology.2008.06.095
- Bergeron, A., M. H. Crete, Y. Brindle, and P. Manjunath. 2004. Low-density lipoprotein fraction from hen's egg yolk decreases the binding of the major proteins of bovine seminal plasma to sperm and prevents lipid efflux from the sperm membrane. Biol. Reprod. 70:708-717. https://doi.org/10.1095/biolreprod.103.022996
- Bowden, R. M., M. A. Ewert, J. L. Lipar, and C. E. Nelson. 2001. Concentrations of steroid hormones in layers and biopsies of chelonian egg yolks. Gen. Comp. Endocrinol. 121:95-103. https://doi.org/10.1006/gcen.2000.7579
- Canisso, I. F., F. A. Souza, J. M. Ortigoza, G. R. Carvalho, M. C. G. Davies-Morel, E. C. Silva, J. D. Guimaraes, and A. L. Lima. 2008. Freezing donkey semen (Equus asinus). Rev. Inv. Vet. Peru 19:113-125.
- Cheng, F. P., A. Fazeli, W. F. Voorhout, A. Marks, M. M. Bevers, and B. Colenbrander. 1996. Use of peanut agglutinin to assess the acrosomal status and the zona pellucida-induced acrosome reaction in stallion spermatozoa. J. Androl. 17:674-682.
- England, G. C. 1993. Cryopreservation of dog semen: A review. J. Reprod. Fertil. Suppl. 47:243-255.
- Gao, D. and J. K. Critser. 2000. Mechanisms of cryoinjury in living cells. ILAR J. 41:187-196. https://doi.org/10.1093/ilar.41.4.187
- Graham, J. K. and R. H. Foote. 1987. Effect of several lipids, fatty acyl chain length, and degree of unsaturation on the motility of bull spermatozoa after cold shock and freezing. Cryobiology 24:42-52. https://doi.org/10.1016/0011-2240(87)90005-8
- Hewitt, D. A., R. Leahy, I. M. Sheldon, and G. C. W. England. 2001. Cryopreservation of epididymal dog sperm. Anim. Reprod. Sci. 67:101-111. https://doi.org/10.1016/S0378-4320(01)00090-2
- Hori, T., M. Ichikawa, E. Kawakami, and T. Tsutsui. 2004. Artificial insemination of frozen epididymal sperm in beagle dogs. J. Vet. Med. Sci. 66:37-41. https://doi.org/10.1292/jvms.66.37
- Jiang, Z. L., Q. W. Li, W. Y. Li, J. H. Hu, H. W. Zhao, and S. S. Zhang. 2007. Effect of low density lipoprotein on DNA integrity of freezing-thawing boar sperm by neutral comet assay. Anim. Reprod. Sci. 99:401-407. https://doi.org/10.1016/j.anireprosci.2006.08.022
- Kampschmidt, R. F., D. T. Mayer, and H. A. Herman. 1953. Lipid and lipoprotein constituents of egg yolk in the resistance and storage of bull spermatozoa. J. Dairy Sci. 36:733-742. https://doi.org/10.3168/jds.S0022-0302(53)91553-7
- Manee-In, S., S. Parmornsupornvichit, S. Kraiprayoon, T. Tharasanit, P. Chanapiwat, and K. Kaeoket. 2014. L-carnitine supplemented extender improves cryopreserved-thawed cat epididymal sperm motility. Asian Australas. J. Anim. Sci. 27:791-796. https://doi.org/10.5713/ajas.2013.13565
- Manjunath, P. and I. Therien. 2002. Role of seminal plasma phospholipid-binding proteins in sperm membrane lipid modification that occurs during capacitation. J. Reprod. Immunol. 53:109-119. https://doi.org/10.1016/S0165-0378(01)00098-5
- Meyers, S. A., J. W. Overstreet, I. K. Liu, and E. Z. Drobnis. 1995. Capacitation in vitro of stallion spermatozoa: Comparison of progesterone-induced acrosome reactions in fertile and subfertile males. J. Androl. 16:47-54.
- Moussa, M., V. Marinet, A. Trimeche, D. Tainturier, and M. Anton. 2002. Low density lipoproteins extracted from hen egg yolk by an easy method: cryoprotective effect on frozen-thawed bull semen. Theriogenology 57:1695-1706. https://doi.org/10.1016/S0093-691X(02)00682-9
- Oliphant, G. and C. A. Singhas. 1979. Iodination of rabbit sperm plasma membrane: Relationship of specific surface proteins to epididymal function and sperm capacitation. Biol. Reprod. 21:937-944. https://doi.org/10.1095/biolreprod21.4.937
- Pena, F. J. 2007. Detecting subtle changes in sperm membranes in veterinary andrology. Asian J. Androl. 9:731-737. https://doi.org/10.1111/j.1745-7262.2007.00311.x
- Platz, C. C., Jr. and S. W. Seager. 1978. Semen collection by electroejaculation in the domestic cat. J. Am. Vet. Med. Assoc. 173:1353-1355.
- Tittarelli, C., C. A. Savignone, E. Arnaudin, M. C. Stornelli, M. A. Stornelli, and R. L. de la Sota. 2006. Effect of storage media and storage time on survival of spermatozoa recovered from canine and feline epididymides. Theriogenology 66:1637-1640. https://doi.org/10.1016/j.theriogenology.2006.01.021
- Varela Junior, A. S., C. D. Corcini, R. R. Ulguim, M. V. F. Alvarenga, I. Bianchi, M. N. Correa, T. Lucia, Jr., and J. C. Deschamps. 2009. Effect of low density lipoprotein on the quality of cryopreserved dog semen. Anim. Reprod. Sci. 115:323-327. https://doi.org/10.1016/j.anireprosci.2008.11.002
- White, I. G. 1993. Lipids and calcium uptake of sperm in relation to cold shock and preservation: A review. Reprod. Fertil. Dev. 5:639-658. https://doi.org/10.1071/RD9930639
- Wistrom, C. A. and S. Meizel. 1993. Evidence suggesting involvement of a unique human sperm steroid receptor/Clchannel complex in the progesterone-initiated acrosome reaction. Dev. Biol. 159:679-690. https://doi.org/10.1006/dbio.1993.1274
- Witte, T. S. and S. Schafer-Somi. 2007. Involvement of cholesterol, calcium and progesterone in the induction of capacitation and acrosome reaction of mammalian spermatozoa. Anim. Reprod. Sci. 102:181-193. https://doi.org/10.1016/j.anireprosci.2007.07.007
- Yanagimachi, R. and A. Bhattacharyya. 1988. Acrosome-reacted guinea pig spermatozoa become fusion competent in the presence of extracellular potassium ions. J. Exp. Zool. 248:354-360. https://doi.org/10.1002/jez.1402480314
- Yanagimachi, R. and N. Usui. 1974. Calcium dependence of the acrosome reaction and activation of guinea pig spermatozoa. Exp. Cell Res. 89:161-174. https://doi.org/10.1016/0014-4827(74)90199-2
- Yu, I., N. Songsasen, R. A. Godke, and S. P. Leibo. 2002. Differences among dogs in response of their spermatozoa to cryopreservation using various cooling and warming rates. Cryobiology 44:62-78. https://doi.org/10.1016/S0011-2240(02)00005-6