Deciphering dose and time dependent effect of supplementing SNP and L-NAME in extender on progressive motility of Murrah bull spermatozoa

V R UPADHYAY; A K ROY; SUJATA PANDITA; DIANA KSHETRIMAYUM; PRIYANKA PATOLIYA; KATHAN RAVAL; RAJU KR DEWRY; SARITA SARITA; SAPNA NATH

doi:10.56093/ijans.v91i11.118126

Authors

V R UPADHYAY ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
A K ROY ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
SUJATA PANDITA ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
DIANA KSHETRIMAYUM ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
PRIYANKA PATOLIYA ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
KATHAN RAVAL ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
RAJU KR DEWRY ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
SARITA SARITA ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
SAPNA NATH ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India

https://doi.org/10.56093/ijans.v91i11.118126

Keywords:

Cryopreservation, Extender, NO, Progressive motility, RNS, ROS

Abstract

Male germ cells have the potential to generate reactive oxygen species (ROS) during different stages of differentiation, laboratory manipulation and during their interaction with dissolved oxygen in extender. These ROS and its subset reactive nitrogen species (RNS) perform a concentration specific bimodal role, favouring oxidative eustress at physiological concentrations. The present study was aimed to investigate the dose and time dependent effect of the most potent RNS, i.e. nitric oxide (NO) compounds, specifically SNP (Sodium nitroprusside) and L-NAME (N-Nitro-L-Arginine Methyl Ester) supplementation in extender on progressive motility. The semen samples were divided into 8 aliquots comprising of various concentrations of SNP (Control, 0.1, 1 and 10 μmol/L) and L-NAME (Control, 1, 10 and 100 μmol/L) and refrigerated for 0, 6, 24 and 48 h. The progressive motility (%) improved significantly particularly with the addition of 1 Î¼M SNP and 10 μM L-NAME as compared to control during all time intervals. Subsequently semen sample was cryopreserved with optimum level of SNP (1μmol/L) and L-NAME (10 μmol/L) from these results by adding it in extender and assessed for progressive motility before cryopreservation and after 24 h, 1 month and 2 months of cryopreservation. It is concluded that the addition of SNP and L NAME in cryopreservation media at 1 μM and 10 μM respectively improved the sperm motility significantly. On further evaluation a significant increase in progressive motility was observed for L-NAME treated extender at 24 h and SNP treated extender at 1 and 2 month interval in comparison to control.

Downloads

Download data is not yet available.

References

1. Balercia G, Moretti S, Vignini A, Magagnini M, Mantero F, Boscaro M and Mazzanti L. 2004. Role of nitric oxide

2. concentrations on human sperm motility. Journal of Andrology 25: 245–49.

3. Cottet Rousselle C, Ronot X, Leverve X and Mayol J F. 2011. Cytometric assessment of mitochondria using fluorescent DOI: https://doi.org/10.1002/cyto.a.21061

4. probes. Cytometry Part A 79: 405–25.

5. de Andrade A F, Arruda R P, Torres M A, Pieri N C, Leite T G, Celeghini E C, Oliveira LZ, Gardes T P, Bussiere M C and

6. Silva D F. 2018. Nitric oxide in frozen-thawed equine sperm: Effects on motility, membrane integrity and sperm capacitation. Animal Reproduction Science 195: 176–84. DOI: https://doi.org/10.1016/j.anireprosci.2018.05.022

7. Digamber D V, Loganathasamy K, Leela V and Kumarasamy P. 2016. Influence of sodium nitroprusside on sperm motility, viability and morphology of frozen thawed buffalo semen. Veterinary Science Research Journal 7: 54–61. DOI: https://doi.org/10.15740/HAS/VSRJ/7.1/54-61

8. Dixit V D and Parvizi N. 2001. Nitric oxide and the control of reproduction. Animal Reproduction Science 65: 1–16. DOI: https://doi.org/10.1016/S0378-4320(00)00224-4

9. Doshi S B, Khullar K, Sharma R K and Agarwal A. 2012. Role of reactive nitrogen species in male infertility. Reproductive Biology and Endocrinology 10: 109. DOI: https://doi.org/10.1186/1477-7827-10-109

10. Dowling D K and Simmons L W. 2008. Reactive oxygen species as universal constraints in life history evolution. Proceedings of the Royal Society B: Biological Sciences 276: 1737–45. DOI: https://doi.org/10.1098/rspb.2008.1791

11. Finkel T. 1998. Oxygen radicals and signaling. Current Opinion in Cell Biology 10: 248–53. DOI: https://doi.org/10.1016/S0955-0674(98)80147-6

12. Francavilla F, Santucci R, Macerola B, Ruvolo G and Romano R. 2000. Nitric oxide synthase inhibition in human sperm

13. affects sperm-oocyte fusion but not zona pellucida binding. Biology of Reproduction 63: 425–29. DOI: https://doi.org/10.1095/biolreprod63.2.425

14. Fraser L, Zasiadczyk £ and Strzezek J. 2010. Interactions of egg yolk lipoprotein fraction with boar spermatozoa assessed with a fluorescent membrane probe. Folia Histochemica et Cytobiologica 48: 292–98. DOI: https://doi.org/10.2478/v10042-010-0013-3

15. Hidalgo M, Rodríguez I and Dorado J. 2006. Influence of staining and sampling procedures on goat sperm morphometry using the Sperm Class Analyzer. Theriogenology 66: 996–1003. DOI: https://doi.org/10.1016/j.theriogenology.2006.02.039

16. Kameshwari D B, Siva A B and Shivaji S. 2003. Inhibition of in vitro capacitation of hamster spermatozoa by nitric oxide synthase inhibitors. Cellular and Molecular Biology (Noisyle-Grand, France) 49: 421–28.

17. Khodaei H, Chamani M, Mahdavi B and Akhondi A A. 2016. Effects of adding sodium nitroprusside to semen diluents on motility, viability and lipid peroxidation of sperm in Holstein bulls. International Journal of Fertility and Sterility 9: 521.

18. Leal A C M S, Caldas-Bussiere M C, de Carvalho C P, Viana K S and Quirino C R. 2009. Role of nitric oxide on quality of freshly ejaculated bull spermatozoa during heparin-induced in vitro capacitation. Animal Reproduction Science 116: 38–49. DOI: https://doi.org/10.1016/j.anireprosci.2008.12.020

19. Lee N P and Cheng C Y. 2009. Nitric oxide and cyclic nucleotides: their roles in junction dynamics and spermatogenesis. Molecular Mechanisms in Spermatogenesis. Springer, New York, NY: 172–85. DOI: https://doi.org/10.1007/978-0-387-09597-4_10

20. Lewis S E , Donnelly E T, Sterling E S, Kennedy M S, Thompson W and Chakravarthy U. 1996. Nitric oxide synthase and nitrite production in human spermatozoa: Evidence that endogenous nitric oxide is beneficial to sperm motility. Molecular Human Reproduction 2: 873–78. DOI: https://doi.org/10.1093/molehr/2.11.873

21. Lone S A, Prasad J K, Ghosh S K, Kumar N, Bhat S A and Das G K. 2016. Effect of cholesterol loaded cyclodextrin on activity of antioxidants during cryopreservation of buffalo (Bubalus bubalis) semen. Indian Journal of Animal Sciences 86: 1255–58. DOI: https://doi.org/10.56093/ijans.v86i11.62993

22. Lone S A, Prasad J K, Ghosh S K, Das G K, Balamurugan B and Verma M R. 2018. Study on correlation of sperm quality

23. parameters with antioxidant and oxidant status of buffalo bull semen during various stages of cryopreservation. Andrologia 50: e12970. DOI: https://doi.org/10.1111/and.12970

24. Miraglia E, De Angelis F, Gazzano E, Hassanpour H, Bertagna A, Aldieri E, Revelli A, Ghigo D 2011. Nitric oxide stimulates human sperm motility via activation of the cyclic GMP/protein kinase G signaling pathway. Reproduction 141: 47–54. DOI: https://doi.org/10.1530/REP-10-0151

25. Mohammed O A, Abdulkareem T A, Ibrahim F F, Al-Zaidi O H, Latif W E and Alwan S H. 2020. Effect of adding pentoxifylline and nitric oxide to tris extender on some post-cryopreserved semen attributes of Holstein bulls. The Iraqi Journal of Agricultural Science 51: 619–28. DOI: https://doi.org/10.36103/ijas.v51i2.989

26. Nash K M, Rockenbauer A and Villamena F A. 2012. Reactive nitrogen species reactivities with nitrones: Theoretical and experimental studies. Chemical Research in Toxicology 25: 1581–97. DOI: https://doi.org/10.1021/tx200526y

27. Naskar S. 2018. ‘Impact of oxidative stress and antioxidants on sperm quality in Murrah bulls’. M.V.Sc. Thesis, ICAR-National Dairy Research Institute, Karnal.

28. Ollero S E, Gil-Guzman E, Lopez M C, Sharma R K, Agarwal A, Larson K, Evenson D, Thomas A J and Alvarez J G. 2001.

29. Characterization of subsets of human spermatozoa at different stages of maturation: Implication in the diagnosis and treatment of male infertility. Human Reproduction 16: 1912–21. DOI: https://doi.org/10.1093/humrep/16.9.1912

30. Panth S. 2017. ‘Reactive nitrogen species as biomarker of semen quality in crossbred cattle bulls’. M.V.Sc. Thesis, ICAR-National Dairy Research Institute, Karnal, Haryana.

31. Raad G, Lteif L, Lahoud R, Azoury J, Azoury J, Tanios J, Hazzouri M and Azoury J. 2018. Cryopreservation media

32. differentially affect sperm motility, morphology and DNA integrity. Andrology 6: 836–45. DOI: https://doi.org/10.1111/andr.12531

33. Rahman M S, Kwon W S, Lee J S, Kim J, Yoon S J, Park Y J and Pang M G. 2014. Sodium nitroprusside suppresses male

34. fertility in vitro. Andrology 2: 899–909.

35. Rodriguez P C, O’flaherty C M, Beconi M T and Beorlegui N B. 2005. Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways. Animal Reproduction Science 85: 231–42. DOI: https://doi.org/10.1016/j.anireprosci.2004.05.018

36. Rosselli M, Dubey R K, Imthurn B, Macas E and Keller P J. 1995. Andrology: Effects of nitric oxide on human spermatozoa: Evidence that nitric oxide decreases sperm motility and induces sperm toxicity. Human Reproduction 10: 786–90. DOI: https://doi.org/10.1093/oxfordjournals.humrep.a136174

37. Saleh R A, Agarwal A, Kandirali E, Sharma R K, Thomas A J, Nada E A, Evenson D P and Alvarez J G. 2002. Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa. Fertility and Sterility 78: 1215–24. DOI: https://doi.org/10.1016/S0015-0282(02)04237-1

38. Sarlos P, Molnar A, Kokai A, Gabor G Y and Ratky J. 2002. Comparative evaluation of the effect of antioxidants in the

39. conservation of ram semen. Acta Veterinaria Hungarica 50: 235–45.

40. Srivastava N and Pande M. 2017. Protocols in Semen Biology (Comparing Assays). Springer Singapore. Srivastava S, Desai P, Coutinho E and Govil G. 2006. Mechanism of action of L-arginine on the vitality of spermatozoa is primarily through increased biosynthesis of nitric oxide. Biology of Reproduction 74: 954–58. DOI: https://doi.org/10.1095/biolreprod.105.046896

41. Upadhyay V R, Roy A K, Pandita S, Dewry R K, Yadav H P, Raval K and Patoliya P. 2021a. Time dependent impact of

42. reactive oxidants on seminal attributes of murrah bull during cryopreservation and storage. Indian Journal of Animal Research B: 4408.

43. Upadhyay V R, Ramesh V, Dewry R K, Kumar G, Raval K and Patoliya P. 2021b. Implications of cryopreservation on

44. structural and functional attributes of bovine spermatozoa: An overview. Andrologia 53: e14154.

45. Wu T P, Huang B M, Tsai H C, Lui M C and Liu M Y. 2004. Effects of nitric oxide on human spermatozoa activity,

46. fertilization and mouse embryonic development. Archives of Andrology 50: 173–79.

47. Zamir N A, Barkan D A, Keynan N U, Naor Z and Breitbart H A. 1995. Atrial natriuretic peptide induces acrosomal exocytosis in bovine spermatozoa. American Journal of Physiology-Endocrinology and Metabolism 269: E216–E221. DOI: https://doi.org/10.1152/ajpendo.1995.269.2.E216