Effect of adding L-glutamine to Tris diluent on cryopreservation of coated sperm of Taleshi ram

Document Type : Research Paper

Authors

1 M.Sc. Student, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan

2 Associate professor, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Rasht

Abstract

Experiment was conducted to evaluate the effect of L-glutamine on coated sperm freezing by using four rams. Semen was collected by artificial vagina contact with a tube containing Tris-fructose-egg yolk 15%. Samples were pooled and centrifuged. After removing supernatant and diluting pellet, samples were split into 5 equal part and added 0, 40, 80, 120 and 160 mM L-glutamine. Samples were frozen by using liquid nitrogen After two weeks, samples were thawed and incubated at 37 °C for nine h. Sperm motility, plasma membrane integrity, viability (Hoechst 33258) and acrosome reaction (PNA-Alexa -flur-488) were investigated at 0, 3, 6 and 9 h. The main effect of L-Glutamine showed that sperm viability was higher in 0 (35.95%), 40 (40.65%) and 80 (45.95%) mM L-Glutamine than 160 mM L-Glutamine (30.40%; P<0.05). There was an interaction between incubation time and L-Glutamine on sperm motility and plasma membrane integrity (P<0.05). After thawing and 3 h later, the highest of plasma membrane integrity was observed in 80 mM L-Glutamine (49.25% and 47.34% respectively; P<0.05). Nine hours after thawing, the highest of sperm motility (26.4%) was observed in 80 mM L-Glutamine (P<0.05). Therefore, the addition of 80 mM L-glutamine to Tris-glycerol diluent (5%) improves coated sperm survival.

Keywords

References

Al Ahmad, M. Z., Chatagnon, G., Amirat‐Briand, L., Moussa, M., Tainturier, D., Anton, M. & Fieni, F. (2008). Use of glutamine and low density lipoproteins isolated from egg yolk to improve buck semen freezing. Reproduction in Domestic Animals, 43, 429-436.
Amirat-Briand, L., Bencharif, D., Vera-Munoz, O., Bel Hadj Ali, H., Destrumelle, S., Desherces, S., Schmidt, E., Anton, M. & Tainturier, D. (2009). Effect of glutamine on post-thaw motility of bull spermatozoa after association with LDL (low density lipoproteins) extender: preliminary results. Theriogenology, 71, 1209-1214.
Aurich, C. (2005). Factors affecting the plasma membrane function of cooled-stored stallion spermatozoa. Animal Reproduction Science, 89, 65-75.
Bencharif, D., Amirat, L., Pascal, O., Anton, M., Schmitt, E., Desherces, S., Delhomme, G., Langlois, M. L., Barriere, P., Larrat, M. & Tainturier D. (2010). The advantages of combining LDL (low density lipoproteins) with glutamine for the cryopreservation of canine semen. Reproduction in Domestic Animals, 45, 189-200.
Bergeron, A. & Manjunath, P. (2006). New insights towards understanding the mechanisms of sperm protection by egg yolk and milk. Molecular Reproduction and Development, 73, 1338-1344.
Bredderman, P. J. & Foote, R. H. (1969). Volume of stressed bull spermatozoa and protoplasmic droplets, and the relationship of cell size to motility and fertility. Journal of Animal Science, 28, 496-501.
Bucak, M. N., Tuncer, P. B., Sarıözkan, S. & Ulutaş, P. A. (2009). Comparison of the effects of glutamine and an amino acid solution on post-thawed ram sperm parameters, lipid peroxidation and anti-oxidant activities. Small Ruminant Research, 81, 13-17.
Cseh, S., Faigl, V. & Amiridis, G.S., 2012. Semen processing and artificial insemination in health management of small ruminants. Anim. Reprod. Sci. 130, 187–92.
de Leeuw, A. M., den Daas, J. H. & Woelders, H. (1991). The fix vital stain method simultaneous determination of viability and acrosomal and status of bovine spermatozoa. Journal of Andrology, 12, 112-118.
de Mercado, E., Hernandez, M., Sanz, E., Rodriguez, A., Gomez, E., Vazquez, J. M., Martinez, E. A. & Roca, J. (2009). Evaluation of L-Glutamine for cryopreservation of boar spermatozoa. Animal Reproduction Science, 115, 149-157.
De Pauw, I. M. C., Van Soom, A., Maes, D., Verberckmoes, S. & de Kruif, A. (2003). Effect of sperm coating on the survival and penetrating ability of in vitro stored bovine spermatozoa. Theriogenology, 59, 1109-1122
Fiser, P. S., Ainsworth, L. & Langford, G. A. (1981). Effect of osmolality of skim-milk diluents and thawing rate on cryosurvival of ram spermatozoa. Cryobiology, 18, 399-403.
Forouzanfar, M., Fazilati, M., Hosseini, S. M., Moulavi, F., Hajian, M., Salehi, S. A., Rabiei, A. & Nasr-Esfahani, M. H. (2007). Investigation of different glycerol and egg yolk concentration on freezing Bakhtiari ram semen. Journal of Iranian Anatomical Sciences, 5, 17-25. (In Farsi)
Hammerstedt, R. H., Graham, J. K. & Nolan, J. P. (1990). Cryopreservation of mammalian sperm: what we ask them to survive. Journal of Andrology , 11, 73-88
Jeyendran, R. S., Van der ven, H. H. & Zaneveld, L. J. (1992). The hypoosmotic swelling test: an update. Archives of Andrology, 29, 105-116.
Khlifaoui, M., Battut, I., Bruyas, J. F., Chatagnon, G., Trimeche, A. & Tainturier, D. (2005). Effects of glutamine on post-thaw motility of stallion spermatozoa: an approach of the mechanism of action at spermatozoa level. Theriogenology, 63, 138-149.
Kruuv, J. & Glofcheski, D. J. (1992). Protective effects of amino acids against freeze-thaw damage in mammalian cells. Cryobiology. 29, 291-295.
Kundu, C. N., Das, K. & Majumder, G. C. (2001). Effect of amino acids on goat cauda epididymal sperm cryopreservation using a chemically defined model system. Cryobiology, 42, 21-27.
Li, Y., Si, W., Zhang, X., Dinnyes, A. & Ji, W. (2003). Effect of amino acids on cryopreservation of cynomologus monkey (Macaca fascicularis) sperm. American Journal of Primatology, 59, 159–165.
Mohammadi-Nohdehy, A. & Roostaei-Ali Mehr, M. (2013). Effect of Egg Yolk and Cooling on Storage of Ram Coated Spermatozoa. Journal of Studies of Animal Sciences Iran, 5, 77-83.
Manjunath, P. & Thérien, I. (2002). Role of seminal plasma phospholipid-binding proteins in sperm membrane lipid modification that occurs during capacitation. Journal of Reproductive Immunology, 53, 109-119.
Mortimer, D. (1994). Sperm recovery techniques to maximize fertilizing capacity. Reproduction, Fertility and Development, 6, 25-31.
Parks, J. E. & Graham, J. K. (1992). Effects of cryopreservation procedures on sperm membranes. Theriogenology, 38,209-222.
Purdy, P. H. (2006). A review on goat sperm cryopreservation. Small Ruminant Research, 63, 215-225.
Renard, P., Grizard, G., Griveau, J. F., Sion, B., Boucher, D. & Le Lannou, D. (1996). Improvement of motility and fertilization potential of post-thaw human sperm using glutamine. Cryobiology, 33, 311–319.
Roostaei-Ali Mehr M.,  Sharafi F. (2013 ) The effect of seminal plasma on the quality of coated ram frozen-thawed spermatozoa. Iranian Journal of Veterinary Research, 14, 305-312.
Salamon, S. & Maxwell, W. M. C. (2000). Storage of ram semen. Animal Reproduction Science, 62, 77-111.
Smith, J. D. & Graham, E. F. (1972). Free amino acids in ram seminal plasma. Journal of Animal Science, 3, 601-604.
Trimeche, A., Renard, P., Le Lannou, D., Barriere, P. & Tainturier, D. (1996). Improvement of motility of post-thaw poitou jackass sperm using glutamine. Theriogenology, 45, 1015–1027.
Trimeche, A., Yvon, J. M., Vidament, M., Palmer, E. & Magistrini, M. (1999). Effects of glutamine, proline, histidine and betaine on post-thaw motility of stallion spermatozoa. Theriogenology, 52, 181–191.
Varisli, O., Uguz, C., Agca, C. & Agca Y. (2009). Motility and acrosomal integrity comparisions between ejaculated and epididimal ram sperm after exposure to a range of anisosmotic solutions, cryoprotective agents and low temperatures. Animal Reproduction Science, 110, 256-268.
Watson, P. F. (1990). Artificial insemination and the preservation of semen. In: Marshall’s Physiology of Reproduction. Reproduction in the male. Lamming G.E, (Ed). Churchill Livingston.(pp, 747–869).
 
Iranian Journal of animal Science
Volume 45, Issue 3 - Serial Number 3
October 2014
Pages 289-296

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History

  • Receive Date: 23 February 2013
  • Revise Date: 08 June 2015
  • Accept Date: 10 October 2013

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