Effect of additives in semen extender on post thaw sperm quality and plasma membrane protein profile of boar semen
327 / 38
Keywords:
Additive, Boar semen, Butylated hydroxytoluene, Cholesterol-loaded methyl-ß-cyclodextrin, Semen, Sperm qualityAbstract
The objective of the study was to evaluate the effect of butylated hydroxytoluene (BHT), cholesterol-loaded methyl-ß-cyclodextrin (CLC), and their combination on sperm motility, viability, plasma membrane integrity and membrane protein profile of frozen-thawed boar semen. Ejaculates (16) from 6 boars were utilized. Each ejaculate was split into 4 fractions and cryopreserved in semen extender containing BHT (0.2 mM), CLC at 5 mg/ 200–240 × 106 sperm, BHT (0.2 mM) plus CLC at 5 mg/ 200–240 × 106 sperm or without BHT and CLC. Semen samples were evaluated for sperm motility, viability (propidium iodide assay), plasma membrane integrity (HOST) and membrane protein profile (SDS-PAGE) after equilibration and after freezing. The supplementation of BHT, CLC, and BHT plus CLC in semen extender significantly improved the viability and plasma membrane integrity after freezing as compared to control. Post thaw sperm motility was also significantly higher in BHT and BHT plus CLC. In addition, the number of sperm plasma membrane protein loss was less in additive treated sperm as compared to that in non-additive treated sperm. Among the additives, BHT plus CLC showed a significant improvement on post thaw sperm motility along with a non-significant improvement on other sperm parameters as compared to BHT and CLC alone. In conclusion, the supplementation of BHT, and CLC in extender before freezing improved the quality of cryopreserved boar semen and, the beneficial effect was more pronounced when BHT was combined with CLC than with BHT and CLC alone.
Downloads
References
Anghel A, Zamfirescu S, Dragomir C, Nadolu D, Elena D and Florica B. 2010. The effects of antioxidants on the cytological parameters of cryopreserved buck semen. Romanian Biotechnological Letters 15 (3): 26–32.
Blanch E, Tomas C, Graham J K and Moce E. 2012. Response of boar sperm to the treatment with cholesterol-loaded cyclodextrins added prior to cryopreservation. Reproduction in Domestic Animal 47: 959–64. DOI: https://doi.org/10.1111/j.1439-0531.2012.01999.x
Cremades T, Roca J, Rodriguez-Martinez H, Abaigar T, Vazquez J M and Martinez E A. 2005. Kinematic changes during the cryopreservation of boar spermatozoa. Journal of Andrology 26 (5):610–18. DOI: https://doi.org/10.2164/jandrol.05028
Crockett E L. 1998. Cholesterol functions in plasma membranes from ectotherms: membrane-specific roles in adaptation to temperature. American Zoology 38: 291–304. DOI: https://doi.org/10.1093/icb/38.2.291
Dhanju C K, Cheema R S and Kaur S P. 2001. Effect of freezing on protein profiles of sperm membrane extracts and seminal plasma of buffalo bulls. Asian-Australasia Journal of Animal Science 14 (12): 1678–82. DOI: https://doi.org/10.5713/ajas.2001.1678
Galantino-Homer H L, Zeng W X, Megee S O, Dallmeyer M, Voelkl D and Dobrinski I. 2006. Effects of 2- hydroxypropylbeta- 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
Harrison R and Vickers S E.1990. Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. Journal of Reproduction and Fertility 88: 343–52. DOI: https://doi.org/10.1530/jrf.0.0880343
Holt W V. 2000. Basic aspects of frozen storage of semen. Animal Reproduction Science 62: 3–22. DOI: https://doi.org/10.1016/S0378-4320(00)00152-4
Jeyendran R S, Van der Ven H H, Perez-Pelaez M, Crabo B G and Zaneveld L J D. 1984. Development of an assay to assess the functional integrity of the human sperm 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
Johnson L A, Aalbers J G and Grooten H J G. 1988. Artificial insemination of swine: Fecundity of boar semen stored in Beltsville TS BTS., Modified Modena (MM), or MR-A and inseminated on one, three and four days after collection. Zuchthygiene 23: 49–55. DOI: https://doi.org/10.1111/j.1439-0531.1988.tb01080.x
Laemmli U K. 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680– 85. DOI: https://doi.org/10.1038/227680a0
Moce E, Blanch E, Tomas C and Graham J K. 2010. Use of cholesterol in sperm cryopreservation: present moment and perspectives to future. Reproduction Domestic Animal 45 (Suppl. 2): 57–66. DOI: https://doi.org/10.1111/j.1439-0531.2010.01635.x
Ollero M, Bescos O, Cebrin-Perez J A and MuiSo-Blanco T. 1998. Loss of plasma membrane proteins of bull spermatozoa through the freezing-thawing process. Theriogenology 49: 547–55. DOI: https://doi.org/10.1016/S0093-691X(98)00006-5
Roca J, Gil M A, Hernandez M, Parrilla I, Vazquez J M and Martinez E A. 2004. Survival and fertility of boar spermatozoa after freeze thawing in extender supplemented with butylated hydroxytoluene. Journal of Andrology 25: 397–405. DOI: https://doi.org/10.1002/j.1939-4640.2004.tb02806.x
Sancho S. Casas I, Ekwall H, Saravia F, Rodriguez-Martinez H, Rodriguez-Gil J E, Flores E, Pinart E M B, Garcia-Gil N, Bassols J, Pruneda A, Bussalleu E, Yeste M and Bonet S. 2007. Effects of cryopreservation on semen quality and the expression of sperm membrane hexose transporters in the spermatozoa of Iberian pigs. Reproduction 134: 111–21. DOI: https://doi.org/10.1530/REP-07-0118
Tomas C, Blanch E, Hernanadez M, Gil M A, Roca J, Vazquez J M, Martinez E A and Moce E. 2011. Treating boar sperm with cholesterol-loaded cyclodextrins widens the sperm osmotic tolerance limits and enhances the in vitro sperm fertilizing ability. Animal Reproduction Science 129: 209–20. DOI: https://doi.org/10.1016/j.anireprosci.2011.12.003
Watson PF. 2000. The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science 60–61: 481–92. Westendorf P, Richter L and Treu H. 1975. Zur Tiefgefrierung von Ebersperma. Labor-und Besamungsergebnisse mit dem Hiilsenberger Pailletten-verfahren. Dtsch Tierarztl Wschr 82: 261–67. DOI: https://doi.org/10.1016/S0378-4320(00)00099-3
Yeste M, Estrada E, Pinart E, Bonet S, Miro J and Rodriguez-Gil J E. 2014. The improving effect of reduced glutathione on boar sperm cryotolerance is related with the intrinsic ejaculate freezability. Cryobiology 68 (2): 251–61. DOI: https://doi.org/10.1016/j.cryobiol.2014.02.004
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2015 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.