پردیس کشاورزی و منابع طبیعی دانشگاه تهران Iranian Journal of animal Science 2008-4773 45 3 2014 09 23 Effect of adding L-glutamine to Tris diluent on cryopreservation of coated sperm of Taleshi ram Effect of adding L-glutamine to Tris diluent on cryopreservation of coated sperm of Taleshi ram 289 296 53786 10.22059/ijas.2014.53786 FA Sanaz Hasanpoor M.Sc. Student, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan Mohammad Roostaei-Ali Mehr Associate professor, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Rasht 0000-0001-7578-467X Mehrdad Mohammadi Associate professor, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Rasht Journal Article 2013 02 23 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 (<em>Hoechst 33258</em>) and acrosome reaction (<em>PNA-Alexa -flur-488</em>) 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. 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 (<em>Hoechst 33258</em>) and acrosome reaction (<em>PNA-Alexa -flur-488</em>) 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. https://ijas.ut.ac.ir/article_53786_b37117001d67ed257e9a0c38be575e10.pdf