Exogenous cholesterol modulates oxidative stress and freezability of mithun spermatozoa
184 / 118
Keywords:
Cholesterol, Free radicals, Freezability, Mithun, SemenAbstract
Effect of cholesterol loaded cyclodextrin (CLC) on improvement of semen quality parameters (SQPs) and deduction of oxidative stresses following cryopreservation in mithun was explored. Total 50 ejaculates were selected out of 105 collected based on preliminary SQPs. Sperm was treated with 1 mg (Gr II) and 2 mg (Gr III) of CLC/ 120×106 spermatozoa and without CLC served as control (Gr I). Diluted semen samples were cryopreserved at ultralow temperature. Frozen thawed samples were evaluated for motility (progressive forward [FPM]; in bovine cervical mucus penetration test [BCMPT] and in computer assisted sperm analyzer [CASA]), viability, total sperm and nuclear abnormality, intactness of plasma membrane and acrosome, intracellular enzymatic leakage and oxidative profile (Malondialdehyde; MDA). Result revealed a significant improvement in motility (FPM, BCMPT and CASA), viability, acrosomal integrity, cholesterol content and reduction of sperm and nuclear abnormalities, leakage of intracellular enzymes and oxidative stressors in 1 mg CLC treated group as compared to control. Moreover, intactness of acrosome and biochemical membrane was protected significantly in extender containing 1 mg CLC. Hence inclusion of mithun spermatozoa with CLC (1 mg/120×106) prior to freezing improved the survivability in cryopreservation. The results clearly indicated the beneficial effects of CLC supplementation on freezability by reducing cryodamage and protecting the spermatozoa integrity.Downloads
References
Alvarez A L, Serres C, Torres P, Crespo F, Mateos E and Gómez- Cuétara C. 2006. Effect of cholesterol-loaded cyclodextrin on the cryopreservation of donkey spermatozoa. Proceedings of the Ninth International Symposium on Equine Reproduction– Equine Reproduction IX, Kerkrade, The Netherlands. Animal Reproduction Science 94: 89–91. DOI: https://doi.org/10.1016/j.anireprosci.2006.04.026
Amann R P and Graham J K. 1992. Spermatozoal function. Equine Reproduction. (Eds) McKinnon A O, Voss J L. Lea and Febiger, Philadelphia. pp. 717–18.
Barth A D and Oko R J. 1989. Preparation of semen for morphological examination, pp. 8–18. Abnormal Morphology of Bovine Spermatozoa. Iowa State University Press, Ames, IA.
Blottner S, Warnke C, Tuchscherer A, Heinen V and Torner H. 2001. Morphological and functional changes of stallion spermatozoa after cryopreservation during breeding and non- breeding season. Animal Reproduction Science 65: 75–88. DOI: https://doi.org/10.1016/S0378-4320(00)00214-1
Desnoyers L and Manjunath P. 1992. Major proteins of bovine seminal plasma exhibit novel interaction with phospholipids. Journal of Biological Chemistry 267: 10149–55. DOI: https://doi.org/10.1016/S0021-9258(19)50212-5
Galantino-Homer H L, Zeng W X, Megee S O, Dallmeyer M, Voelkl D and Dobrinski I. 2006. Effects of 2-hydroxypropyl-
beta-cyclodextrin and cholesterol on porcine sperm viability and capacitation status following cold shock or incubation. Molecular Reproduction and Development 73: 638–50. DOI: https://doi.org/10.1002/mrd.20437
Hammerstedt R H, Graham J K and Nolan J P. 1990. Cryopreservation of mammalian sperm: what we ask them to survive. Journal of Andrology 11: 73–88.
Jeyendran R S, Vander Ven H H, Parez-Pelaez M, Crabo B G and Zaneweld L J D. 1984. Development of an assay to assess the functional integrity of the human membrane and its relationship to other semen characteristics. Journal of Reproduction and Fertility 70: 219–28. DOI: https://doi.org/10.1530/jrf.0.0700219
Karunakaran M, Dhali A, Mech A, Khate K, Rajkhowa C and Mishra D P. 2007. Preservation of mithun (Bos frontalis) semen at refrigeration temperature. Animal Reproduction Science 101: 257–64. DOI: https://doi.org/10.1016/j.anireprosci.2006.09.011
Kumar A. 2012. ‘Studies on effect of cholesterol loaded cyclodextrin on freezability and in vitro fertility of buffalo spermatozoa’. PhD Thesis, ICAR-Indian Veterinary Research Institute, Uttar Pradesh, India.
Lasley J F and Bogart R. 1944. A comparative study of epididymal and ejaculated spermatozoa of boar. Journal of Animal Science 3: 360–70. DOI: https://doi.org/10.2527/jas1944.34360x
Leahy T and Gadella B M. 2011. Sperm surface changes and physiological consequences induced by sperm handling and storage. Reproduction 142: 759–78. DOI: https://doi.org/10.1530/REP-11-0310
Lee Y S, Lee S, Lee S H, Yang B K and Park C K. 2015. Effect of cholesterol-loaded-cyclodextrin on sperm viability and acrosome reaction in boar semen cryopreservation. Animal Reproduction Science 159: 124–30. DOI: https://doi.org/10.1016/j.anireprosci.2015.06.006
Li G, Saenz J, Godke R A and Devireddy R V. 2006. Effect of glycerol and cholesterol-loaded cyclodextrin on freezing- induced water loss in bovine spermatozoa. Reproduction 131: 875–86. DOI: https://doi.org/10.1530/rep.1.00995
Matouseket J, Riha J, Sarsen V, Veselky H and Londa F. 1989. Penetration of cervical mucus and other body fluids by bull sperm in capillary tubes. Animal Reproduction Science 18: 161– 66. DOI: https://doi.org/10.1016/0378-4320(89)90018-3
Moce E, Purdy P H and Graham J K. 2010. Treating ram sperm with cholesterol–loaded cyclodextrins improves cryosurvival. Animal Reproduction Science 118: 236–47. DOI: https://doi.org/10.1016/j.anireprosci.2009.06.013
Mondal M, Karunakaran M, Lee K B and Rajkhowa C. 2010. Characterization of mithun (Bos frontalis) ejaculates and fertility of cryopreserved sperm. Animal Reproduction Science 118: 210–16. DOI: https://doi.org/10.1016/j.anireprosci.2009.09.001
Moore A I, Edward L, Squires J and Graham K. 2005. Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology 51: 241–49. DOI: https://doi.org/10.1016/j.cryobiol.2005.07.004
O’Connell M, McClure N and Lewis S E. 2002. The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Human Reproduction 17: 704–09. DOI: https://doi.org/10.1093/humrep/17.3.704
Ozkavukcu S, Erdemli E, Isik A, Oztuna D and Karahuseyinoglu S. 2008. Effects of cryopreservation on sperm parameters and ultrastructural morphology of human spermatozoa. Journal of Assisted Reproduction and Genetics 25(8): 403–11. DOI: https://doi.org/10.1007/s10815-008-9232-3
Paasch U, Grunewald S, Fitzl G and Glander H J. 20003. Deterioration of plasma membrane is associated with activated caspases in human spermatozoa. Journal of Andrology 24: 246–52. DOI: https://doi.org/10.1002/j.1939-4640.2003.tb02669.x
Parks J E and Graham J K. 1992. Effects of cryopreservation procedures on sperm membranes. Theriogenology 38: 209– 22. DOI: https://doi.org/10.1016/0093-691X(92)90231-F
Perumal P. 2014. ‘Studies on effect of low density lipoprotein on freezability and fertilizing ability of spermatozoa in mithun (Bos frontalis) bulls’. Ph.D. Thesis, ICAR-Indian Veterinary Research Institute, Uttar Pradesh, India.
Perumal P. 2014. Computer-Assisted sperm analysis of freezable and non-freezable mithun (Bos frontalis) semen. Journal of Animal Science. Article ID 675031. DOI: https://doi.org/10.1155/2014/675031
Presicce G A, Révay T, Nagy S Z, Dinnyés A and Kovács A. 2003. Complex staining of water buffalo (Bubalus bubalis) spermatozoa. Bubalus Bubalis 2: 55–60.
Purdy P H and Graham J K. 2004. Effect of adding cholesterol to bull sperm membranes on sperm capacitation, the acrosome reaction, and fertility. Biology of Reproduction 71: 522–27. DOI: https://doi.org/10.1095/biolreprod.103.025577
Rajoriya J S, Prasad J K, Ramteke S S, Perumal P, Ghosh S K, Singh M, Pande M and Srivastava N. 2016. Enriching membrane cholesterol improves stability and cryosurvival of buffalo spermatozoa. Animal Reproduction Science 164: 72– 81. DOI: https://doi.org/10.1016/j.anireprosci.2015.11.014
Salisbury G W, VanDemark N L and Lodge J R. 1985. Physiology of Reproduction and Artificial Insemination of Cattle. 2nd edn. WH Freeman and Company, New York, pp. 268–74.
Salmon V M, Leclerc P and Bailey J L. 2017. Novel technical strategies to optimize cryopreservation of goat semen using cholesterol-loaded cyclodextrin. Cryobiology 74: 19–24. DOI: https://doi.org/10.1016/j.cryobiol.2016.12.010
Srivastava N, Srivastava S K, Ghosh S K, Prasad J K, Kumar A and Tripathi R P. 2012. Effect of sequestration of PDC-109 protein on freezabilty and fertilizing ability of crossbred bull spermatozoa during cryopreservation. Animal Reproduction Science 131: 54–62. DOI: https://doi.org/10.1016/j.anireprosci.2012.02.003
Tomar N S. 1997. Artificial Insemination and Reproduction of Cattle and Buffaloes. Sarojprakashan, Allahabad, India.
Walters E M, Men H, Agca Y, Mullen S F, Critser E S and Critser J K. 2005. Osmotic tolerance of mouse spermatozoa from various genetic backgrounds: acrosome integrity, membrane integrity, and maintenance of motility. Cryobiology 50: 193– 205. DOI: https://doi.org/10.1016/j.cryobiol.2005.01.004
Watson P F. 1975. Use of Giemsa stain to detect change in acrosome of frozen ram spermatozoa. Veterinary Record 97: 12–15. DOI: https://doi.org/10.1136/vr.97.1.12
Watson P F. 1981. The effect of cold shock on sperm cell membrane. Effects of Low Temperature on Biological Membranes. (Eds) Morris G J and Clarke A. Academic Press, London. pp. 189–218.
Yadav H P, Anuj Kumar, Shah N, Chauhan D S, Saxena A and Yadav S. 2017. Effect of cholesterol loaded cyclodextrin supplementation on tyrosine phosphorylation and apoptosis like changes in frozen thawed Hariana bull spermatozoa. Theriogenology 96: 164–71. DOI: https://doi.org/10.1016/j.theriogenology.2017.04.016
Zahn F S, Papa F O and Dell’Acqua Jr. J A. 2002. Cholesterol incorporation on equine sperm membrane: Effects on post-thaw sperm parameters and fertility. Theriogenology 58: 237–40. DOI: https://doi.org/10.1016/S0093-691X(02)00762-8
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2019 The Indian Journal of Animal Sciences

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The copyright of the articles published in The Indian Journal of Animal Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.