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http://dx.doi.org/10.5713/ajas.2010.90387

The Effects of Different Concentrations of Glycine and Cysteine on the Freezability of Moghani Ram Spermatozoa  

Khalili, B. (Jafar-Abad Livestock Central Research Institute, Ministry of Agriculture)
Jafaroghli, M. (College of Agriculture, University of Payam-e- Noor)
Farshad, Abbas (Department of Animal Science, College of Agriculture, University of Kurdistan)
Paresh-Khiavi, M. (Jafar-Abad Livestock Central Research Institute, Ministry of Agriculture)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.23, no.3, 2010 , pp. 318-325 More about this Journal
Abstract
Two experiments were designed to evaluate the effects of the amino acids glycine and cysteine on cryopreservation of ram spermatozoa. After primary evaluation of collected ejaculates, the semen samples were pooled and diluted 1:4 before cooling (experiment 1) and freezing (experiment 2) with Tris-Citrate-Fructose-Yolk (TCFY) extender supplemented with different concentrations of glycine and cysteine (5, 10, 15 and 20 mM). As the control, semen was diluted and frozen in the extender without amino acids. Motility, viability and membrane integrity were assessed as the parameters for semen quality in the first experiment. In the second experiment, motility, progressive motility, viability, membranes and acrosome integrity were evaluated after the freezing-thawing process. The results of the first experiment indicated that the addition of 10 and 15 mM cysteine compared to the control (basic) extender significantly increased (p<0.01) the motility, viability and membrane integrity of spermatozoa after cooling. However, further increasing these amino acids up to 20 mM had a significant negative effect (p<0.05). Our results showed no significant differences (p>0.05) between 5 mM glycine compared to 5 mM cysteine and between 20 mM glycine and 20 mM cysteine. The results of experiment 2 showed that the amino acids significantly improved post-thaw motility, progressive motility, viability, membranes and acrosome integrity of ram spermatozoa. These positive effects were observed at concentrations between 5 to 15 mM of glycine and cysteine, with the best results at 15 mM. Further increasing of amino acid concentrations significantly decreased the post-thaw characteristics of spermatozoa, but the results showed that cysteine was better than glycine and control extenders. The data indicated that addition of glycine or cysteine to the freezing extender can be recommended for cryopreservation of ram spermatozoa. However, further studies are still needed to determine the effect of such addition on fertility in farm animals.
Keywords
Cryopreservation; Glycine; Cysteine; Moghani Ram Spermatozoa;
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1 Billard, R. and Y. Menezo. 1984. The amino acids composition of rainbow trout seminal fluid and blood plasma: a comparison with carp. Aquaculture 41:255-258   DOI   ScienceOn
2 Khlifaouia, M., I. Battuta, J. F. Bruyasa, G. Chatagnona, A. Trimecheb and D. Tainturiera. 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   DOI   ScienceOn
3 Lalonde, R., J. Lepock and J. Kruuv. 1991. Site of freeze-thaw damage and cryopreservation by amino acids of the calcium ATPase of the sarcoplasmic reticulum. Biochem. Biophys. Acta. 1079:128-138   DOI   ScienceOn
4 Morris, G. I., G. I. Morris and A. Clarke. 1981. Effects of low temperature on biological membranes. New York, Academic Press, 241-377
5 Hammerstedt, R. H., J. K. Graham and J. P. Nolan. 1990. Cryopreservation of mammalian sperm: what we ask them to survive. J. Androl. 11:73-88   PUBMED
6 Bailey, J. L., J. F. Bilodeau and N. Cormier. 2000. Semen cryopreservation in domestics animals: a damaging and capaciting phenomenon. J. Androl. 21:1-7   PUBMED
7 Aitken, R. J. and H. Fisher. 1994. Reative oxygen species generation and human spermatozoa: the balance of benefit and risk. Bio-assays. 16:259-267
8 Ali Al Ahmad, M. Z., G. Chatagnon, L. Amirat-Briand, M. Moussa, D. Tainturier, M. Anton and F. Fieni. 2008. Use of glutamine and low density lipoproteins isolated from egg yolk to improve buck semen freezing. Reprod. Domest. Anim. 43:429-436   DOI   PUBMED   ScienceOn
9 Alvarez, J. G. and B. T. Storey. 1983. Taurine, hypotaurine, epinephrine and albumin inhibit lipid peroxidation in rabbit spermatozoa and protect against loss of motility. Biol. Reprod. 29:548-555   DOI   ScienceOn
10 Bucak, M. N., A. Atessahin and Y. Abdurrauf. 2008. Effect of antioxidants and oxidative stress parameters on ram after the freeze-thawing process. Small Rumin. Res. 75:128-134   DOI   ScienceOn
11 Darin-Bennett, A. and I. G. White, 1977. Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock. Cryobiology 14: 466-470   DOI   ScienceOn
12 Dhami, A. J. and K. L. Shani. 1994. Evaluation of different cooling rates, equilibration periods and diluents for effects on deep-freezing, enzyme leakage and fertility of taurine bull spermatozoa. Theriogenology 40:1269-1280   DOI   ScienceOn
13 Purdy, P. H. 2006. A review on goat sperm cryopreservation. Small Rumin. Res. 63:215-225   DOI   ScienceOn
14 Evans, G. and W. M. S. Maxwell. 1987. Salamon's artificial insemination of sheep and goats. University Press, Sydney, NSW, Australia
15 Fiser, P. S. and R. W. Fairful. 1989. The effect of glycerol related osmotic changes on post thaw motility and acrosomal integrity of ram spermatozoa. Cryobiology 26:64-69   DOI   ScienceOn
16 Flipse, R. J. and J. Q. Almquist. 1956. Diluters for bovine semen. IX. Motility of bovine spermatozoa in milk-glycine and egg yolk-glycine diluents with and without glycerol, in: J. Pennsylv. Agric. Exp. Station. 1690-1696
17 Rudolph, A. S. and J. H. Crowe. 1985. Membraned stabilization during freezing: the role of two natural cryoprotectants, trehalose and proline. Cryobiology 22:367-377   DOI   ScienceOn
18 Sheshtawy, R. I., G. A. El-Sisy and W. S. El-Nattat. 2008. Use of selected amino acids to improve buffalo bull semen cryopreservation. Global Vet. 2:146-150
19 Trimèche, A., J. M. Yvon, M. Vidament, E. Palmer and M. Magistrini. 1999. Effects of glutamine, proline, histidine and betaine on post-thaw motility of stallion spermatozoa, Theriogenology 52:181-191   DOI   ScienceOn
20 Watson, P. F. 2000. The causes of reduced fertility with cryopreserved semen. Anim. Reprod. Sci. 61:481-492   DOI   PUBMED   ScienceOn
21 Watson, P. F. and I. C. A. Martin. 1975. Effects of egg yolk, glycerol and the freezing rate on the viability and acrosomal structures of frozen ram spermatozoa. Aust. J. Biol. Sci. 28: 153-159   PUBMED   ScienceOn
22 Moore, A. I., E. L. Squires and J. K. Graham. 2005. Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology 51:241-249   DOI   ScienceOn
23 Salamon, S. and W. M. Maxwell. 2000. Storage of ram semen. Anim. Reprod. Sci. 62:77-111   DOI   PUBMED   ScienceOn
24 Weitze, K. F. 1977. Untersuchungen zur Tiefgefriekonservierung von Kaninchensperma. Habil- Schr., Tieraerztl. Hochs. Hannover, Germany
25 Baumber, J., B. A. Ball, C. G. Gravance, V. Medina and M. C. G. Davies-Morel. 2000. The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential and membrane lipid peroxidation. J. Androl. 21:895-902   PUBMED   ScienceOn
26 Eiman, M.-E. A. and T. Terada. 2004. Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol. Reprod. 69:1245-1250   DOI   ScienceOn
27 Kumar, S., J. D. Millar and P. F Watson. 2003. The effect of cooling rate on the survival of cryopreserved bull, ram and boar spermatozoa: a comparison of two controlled-rate cooling machines. Cryobiology 46:246-253   DOI   ScienceOn
28 Watson, P. F. 1981. The role of lipid and protein in the proteiction of ram spermatozoa at 5 degrees C by egg-yolk lipoprotein. J. Reprod. Fertil. 62:483-492   DOI   ScienceOn
29 Morris, G. I. and A. Clarke. 1987. Cell at low temperatures. In: The effects of low temperatures on biological systems (Ed. B. W. W. Grout and G. J. Morris). London, Edward Arnold, 72-119
30 Flipse, R. J. and J. Q. Almquist. 1955. Effect of milk-glycine and egg yolk-glycine diluents on the survival of bovine spermatozoa. J. Anim. Sci. 14:1182
31 Uysal, O. and M. N. Bucak. 2007. Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozen-thawed ram semen. Acta Vet. Brno 76:383-390   DOI
32 Atessahin, A., M. N. Bucak, P. B. Tuncer and M. Kizil. 2008. Effect of antioxidant additives on microscopic and oxidative parameters of Angora goat semen following the freeze-thawing process. Small Rumin. Res. 77:38-44   DOI   ScienceOn
33 Matsuoka, T., H. Imai, H. Kohno and Y. Fukui. 2006. Effect of bovine serum albumin and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa. J. Reprod. Dev. 52:675-683   DOI   ScienceOn
34 Kundu, C. N., K. Das and G. C. Majumder. 2001. Effect of amino acids on cauda epididymal sperm cryopreservation using a chemically defined model system. Cryobiology 41:21-27   DOI   ScienceOn
35 SAS, 1996. SAS/STAT Software: Changes and Enhancements through Release 6.12. SAS Institute Inc., Cary, NC, USA
36 Anchordoguy, T., J. F. Carpenter, S. H. Loomis and J. H. Crowe. 1988. Mechanisms of interaction of amino acids with phospholipid bilayers during freezing. Biochim. Biophys. Acta. 946:299-306   DOI   ScienceOn
37 Sinha, M. P., A. K. Sinha, B. K. Singh and P. L. Prasad. 1996. The effect of glutathione on the motility, enzyme leakage and fertility of frozen goat semen. Theriogenology 41:237-243
38 Curry, M. R., J. D. Millar and P. F. Watson. 1994. Calculated optimal cooling rates for ram and human sperm cryopreservation fail to confirm with empirical observations. Biol. Reprod. 51:1014-1021   DOI   ScienceOn
39 Drobnis, E. Z., L. M. Crowe and T. Berger. 1993. Cold shock damage is due to lipid phase-transitions in cell membranes:a demonstration using sperm as a model. J. Exp. Zool. 265:432-437   DOI   ScienceOn
40 Noguchi, S. and J. Matsmuto. 1971. Studies on the control of the denaturation of the fish muscle proteins during frozen storage II. Preventing effect of amino acids and related compounds. Bull. Jap. Soc. Sci. Fish. 37:1115-1122   DOI
41 Iqbal, N. and A. G. Hunter. 1991. Comparison of bovine sperm capacitations systems. J. Dairy Sci. 74:228
42 Purdy, P. H., M. H. Fox and J. K. Graham. 2005. The fluidity of chinese hamster ovary cell and bull sperm membranes after cholesterol addition. Cryobiology 51:102-112   DOI   ScienceOn
43 Chu, T. M., D. Aspinall and L. G. Paleg. 1974. Stress metabolism: Part 6. Temperature stress and the accumulation of proline in barley and radish. Aust. J. Plant Physiol. 1:87-97   DOI
44 Farshad, A., B. Khalili and P. Fazeli. 2009. The effect of different concentrations of glycerol and DMSO on viability of Markhoz goat spermatozoa during different freezing temperatures steps. Pak. J. Biol. Sci. 12:239-245   DOI   ScienceOn
45 Foote, R. H. 1970. Fertility of bull semen at high extension rates in Tris Buffered Extenders. J. Dairy Sci. 53:1475-1477   DOI
46 Lahnsteiner, F., T. Weismann and R. A. Patzner. 1992. Fine structural changes in spermatozoa of the grayling, Thymallus thymallus (Pisces:Teleostei), during routine cryopreservation. Aquaculture 103:73-84   DOI   ScienceOn
47 Bilodeau, J. F., S. Blanchette, I.C. Gagnon and M. A. Sirard. 2001. Thiols prevent $H_2O_2$ - mediated loss of sperm motility in cryopreserved bull semen. Theriogenology 56:275-286   DOI   ScienceOn
48 Li, Y., W. Si, X. Zhang, A. Dinnyes and W. Ji. 2003. Effect of amino acids on cryopreservation of cynmolgus monkey (macaca fascicularis) sperm. Am. J. Primatol. 59:159-165   DOI   ScienceOn
49 Revell, S. G. and R. A. Mrode. 1994. An osmotic resistance test for bovine semen. Anim. Reprod. Sci. 36:77-86   DOI   ScienceOn
50 Dhami, A. J., K. L.Shani and G. Mohan. 1995. Role of prefreeze cooling/holding times, dilutors and thaw rates in improving freezability and post thaw survival of bovine spermatozoa. Ind. Anim. Reprod. 16:117-120
51 Kruuv, J. and D. J. Glofcheski. 1992. Protective effect of amino acids against freeze-thaw damage in mammalian cells. Cryobiology 29:291-295   DOI   ScienceOn
52 Szczesniak-Fabianczyk, B., M. Bochenek, Z. Smorag and M. A. Silvestre. 2006. Effect of antioxidants added to boar semen extender on the semen survival time and sperm chromatin structure. Reprod. Biol. 3:81-87   ScienceOn
53 Gadella, B. M., R. Rathi, J. F. H. M. Brouwers, T. A. E. Stout and B. Colenbrander. 2001. Capacitation and the acrosome reaction in equine sperm. Anim. Reprod. Sci. 68:249-265   DOI   ScienceOn
54 He, S. and L. C. Woods. 2003. Effect of glycine and alanine on short term storage and cryopreservation of striped bass (Morone saxatilis) spermatozoa. Cryobiology 46:17-25   DOI   ScienceOn
55 Roy, A. and M. W. H. Bishop. 1954. Effect of glycine on the survival of bull spermatozoa in vitro. Nature 174:746-747   DOI   ScienceOn
56 Scanchez-Partidata, L. G., W. M. C. Maxwell, L. G. Paleg and B. P. Setchell. 1992. Proline and glycine betaine in cryoprotective diluents for ram spermatozoa. Reprod. Fertil. Dev. 4:113-118   DOI   ScienceOn