Loading...

Proteins and Heparin Binding Proteins of Spermatozoa and Seminal Plasma in Beetal Bucks: Purification, Characterization and Role in In vivo Fertility

DOI: 10.18805/ijar.B-3974    | Article Id: B-3974 | Page : 394-400
Citation :- Proteins and Heparin Binding Proteins of Spermatozoa and Seminal Plasma in Beetal Bucks: Purification, Characterization and Role in In vivo Fertility.Indian Journal of Animal Research.2021.(55):394-400
Navjot S. Dhillon, Ranjna S. Cheema, Sumit Singhal ranjna.cheema@gmail.com
Address : Department of Veterinary Gynecology and Obstetrics, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141 004, Punjab, india.
Submitted Date : 25-12-2019
Accepted Date : 12-06-2020

Abstract

Background: The proteome analysis of seminal plasma and spermatozoa is of special relevance in livestock. Heparin binding proteins (HBPs) found in the seminal plasma of several mammals are shown to bind to sperm membrane and affect a series of events that contribute to normal fertility, such as sperm capacitation, formation of the oviduct reservoir and binding to the oocyte. Profiles of HBPs from seminal plasma and sperm membranes have been associated with sperm fertility. Although, HBPs present in the SP are described in several species, but little is known about HBPs in buck. 
Methods: Seminal plasma (SP) and sperm extracts (SE) of 13 bucks were subjected to heparin-sepharose affinity chromatography. Sperm extract, seminal plasma and purified HBPs and Non-HBPs were fractionated by SDS-PAGE. Total 78 females (6 per buck) were mated with 13 bucks. Bucks were divided into two groups, G-I (high fertile, 83.3-100% FR) and G-II (low fertile, 50-66.7% FR). Relationship between HBPs and fertility rate was observed. 
Result: SDS – PAGE of SP and SE resulted in resolution of 22 (10-240 kDa) and 21 (10-270 kDa) bands, respectively. Based on fertility rate 15 and 13 kDa proteins were absent in SP of higher number of GI-compared to G-II bucks. Fourteen bands ranging from 10 – 180 kDa and 10 – 150 kDa were separated from SP-NHBP and SP-HBP. SP-HBPs of 75, 35, 30, 28, 25 and 13 kDa were present in higher (28.6%, 42.5%, 26.2%, 40.5%, 14.3% and 36.2%) number of high fertile than low fertile bucks. NHBP and HBP purified from SE resolved into 11 bands ranging from 10 – 135 kDa and 10 – 120 kDa, respectively. SE-HBP of 53 kDa, 50/45 kDa and 25 kDa were present in higher percentage of high fertile than low fertile bucks. 

Keywords

Beetle buck Fertility HBP Seminal plasma Spermatozoa

References

  1. Arangasamy, A., Singh, L.P., Ahmed, N., Ansari, M.R., Ram, G.C. (2005). Isolation and characterization of heparin and gelatin binding buffalo seminal plasma proteins and their effect on cauda epididymal spermatozoa. Animal Reproduction Science. 90: 243-254.
  2. Bellin, M.E., Hawkins, H.E., Ax, R.L. (1994). Fertility of range beef bulls grouped according to presence or absence of heparin binding proteins in sperm membranes and seminal fluid. Journal of Animal Science. 72: 2441-2448.
  3. Bellin, M.E., Oyarzo, J.N., Hawkins, H.E., Zhang, H., Smith, R.G., Forrest, D.W., Sprott, L.R., Ax, R.L. (1998). Fertility-associated antigen on bull sperm indicates fertility potential. Journal of Animal Science. 76: 2062-2069.
  4. Calvete, J.J., Raida, M., Gentzel, M., Urbanke, C., Sanz, L., Topfer-Petersen, E. (1997). Isolation and characterization of heparin and phosphoryl choline binding proteins of boar and stallion seminal plasma: primary structure of porcine pB1. FEBS Letters. 407: 201-206.
  5. Cheema, Ranjna S., Bansal, A.K., Patel, M., Gandotra, V.K. (2016). Purification and Characterization of Heparin Binding Proteins from Seminal Plasma of Crossbred Cattle Bulls by Affinity Chromatography, SDS-PAGE and Mass Spectrometery. Journal of Proteomics and Enzymology. 5:1-6.
  6. de Souza, F.F., Martins, M.I., dos Santos, Fernandes, C.E., Ribolla, P.E., Lopes, M.D. (2006). Herapin-binding proteins of canine seminal plasma. Theriogenology. 66: 1606-1609. 
  7. Deori, S., Deka, B., Biswas, R., Nahardeka, N., Arangasamy, A., Bhuyan, D., Kalita, D., Borah, R., Phookan, A. (2018) Characteristics and freezability of Assam Hill goat semen. Indian Journal of Animal Research. 52: 25-28.
  8. Duncan, M.W. and Thompson, H.S. (2007). Proteomics of semen and its constituents. Proteomics Clinical Applications. 1: 861-875.
  9. Dhillon, N. S., Cheema, Ranjna S., Kaswan, S. and Singhal, S. (2019). Seminal attributes, antioxidant defense system and fertility rate reveal an association in Beetal goat bucks. International Journal of Livestock Research. Accepted.
  10. Farooqui, A.A. (1980). Purification of enzymes by heparin-sepharose affinity chromatography. Journal of Chromatography. 184: 335-45.
  11. Fernandes, C.E., DeSouza, F.F., Souza-Neto, J.A., Ribola, P.E.M. (2009). Heparin binding proteins of seminal plasma in Nellore bulls. Ciência Rural. 39: 20-26.
  12. Gupta, N., Cheema, Ranjna S., Singh, A.K. (2019). Characterization of sperm heparin binding proteins (HBPs) using polyclonal antibodies raised against seminal plasma HBPs: Application in buffalo fertility. Indian Journal of Biotechnology. 18: 26-33.
  13. La Falci, V.S.N., Tortorella, H., Rodrigues, J.L., Brandelli, A. (2002). Seasonal variation of goat seminal plasma proteins. Theriogenology. 57: 1035-1048.
  14. Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature. Veterinary World. 227: 680-685.
  15. Lenz, R.W., Martin, J.L., Bellin, M.E. and Ax, R.L. (1988). Predicting fertility of dairy bulls by inducing acrosome reactions in sperm with chondroitin sulphates. Journal of Dairy Science. 71: 1073-1077. 
  16. Lowry, O.H., Rosenberg, N.J., Farr, A.L., Randall, R.J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry. 193: 265-275.
  17. Manaskova, P., Liberda, J., Ticha, M., Jonakova V. (2002). Isolation of nonheparin-binding and heparin-binding proteins of boar prostate. Journal of Chromatography. 770: 137-143.
  18. Marks, J.L. and Ax, R.L. (1985). Relationship of non-return rates of dairy bulls to binding affinity of heparin to sperm. Journal of Dairy Science. 68: 2078-2082.
  19. Martin, J.A.M., Souza, C.E.A., Silva, F.D.A., Cadavida, V.G., Nogueirac, F.C., Domontc, G.B., Oliveirab, J.T., Moura, A.A. (2013). Major heparin-binding proteins of the seminal plasma from Morada Nova rams. Small Ruminanr Research. 114: 115-127.
  20. Mor, V., Das, T., Ray, K., Chatterjee, T. (2006). A heparin-binding membrane protein from goat spermatozoa immobilizes spermatozoa in the presence of complement. Fertility and Sterility. 85: 1142-1149.
  21. Moura, A.A., Chapman, D.A., Koc, H., Killian, G.J. (2007). A comprehensive proteomic analysis of the assessor sex glands fluid from mature Holstein bulls. Animal Reproduction Science. 98: 169-188.
  22. Nandi, P., Ghosh, S., Jana, K., Sen, P.C. (2012). Elucidation of the involvement of p14, a sperm protein during maturation, capacitation and acrosome reaction of caprine spermatozoa. PloS one. 7: e30552.
  23. Nauc, V. and Manjunath, P. (2000). Radio immunoassays for bull seminal plasma proteins (BSP-A1/A2, BSP-A3 and BSP-30-kilo Daltons) and their quantification in seminal plasma and sperm. Biology of Reproduction. 63: 1058-1066.
  24. Schoneck, C., Braun, J., Einspanier, R. (1996). Sperm viability is influenced in vitro by the bovine seminal protein aSFP: effects on motility, mitochondrial activity and lipid peroxidation. Theriogenology. 45: 633-642.
  25. Singh, A.K., Brar, P.S., Cheema, Ranjna S. (2016). Identification of heparin binding proteins in frozen-thawed capacitated/acrosome-reacted spermatozoa and their relationship with fertility of buffalo bull semen. International Journal of Advances in Veterinary Sciences and Technology. 5: 211-216.
  26. Singh, M., Ghosh, S.K., Prasad, J.K., Kuma, R.A., Tripathi, R.P. (2014). Seminal PDC-109 protien vis a vis cholesterol content and freezability of buffalo spermatozoa. Animal Reproduction Science. 141: 22-29.
  27. Sprott, L.R., Forrest, D.W., Zhang, H.M., Dyarzo, J.N., Bellin, M.E., Ax, R.L. (2000). Artificial inseminations outcome in beef females using bovine sperm with a detectable fertility associated antigen. Journal of Animal Science. 78: 795-798.
  28. Srivastava, N., Srivastava, S.K., Ghosh, S.K., Singh, L.P., Prasad, J.K., Kumar, A., Perumal, P., Jerome, A., Thamizharasan, A. (2012). Sequestration of PDC-109 protein improves freezability of crossbred bull spermatozoa. Animal Reproduction Science. 131: 54-62.
  29. Teixeira, D.I.A., Melo, L.M., Gadelha, C., Cunha, A.A., Bloch, R.M.S., Rádis-Baptista, C., Cavada, G., Freitas, B. (2006). Ion-exchange chromatography used to isolate a spermadhesin-related protein from domestic goat (Capra hircus) seminal plasma. General Molecular Research. 5: 79-87.
  30. Villemure, M., Lazure, C., Manjunath, P. (2003). Isolation and characterization of gelatin-binding proteins from goat seminal plasma. Reproductive Biology and Endocrinology. 1: 39-48. 

Global Footprints