Characterization of proteins in cryopreserved and non-cryopreserved seminal plasma of dairy bulls of dif-fering fertility


Seminal plasma is composed of secretions from accessory sex glands, which are mixed with sperm at ejaculation and contribute the majority of semen volume. Seminal plasma is considered a transport and support medium for sperm in the female reproductive tract. Because seminal plasma is not required for fertilization, the importance of its constituents to the establishment of normal pregnancy has been overlooked. Four seminal plasma proteins, Osteopontin, Sper-madhesin Z13, BSP 30 kDa and Phospholipase A2, have been identified as markers of fertility in dairy bulls (Cancel et al., 1997; Moura et al., 2006, 2007). The objective of the present study was to characterize the expression patterns of these proteins and other proteins found to be of interest in seminal plasma of cryopreserved and non-cryopreserved bull semen. Seminal plasma samples were obtained from 16 mature Hol-stein-Friesian bulls at Select Sires Inc. Samples were divided into two groups based on assigned fertility score expressed as the percentage point deviation (PD) of the bull’s non-return rate (NRR) from the average NRR of all bulls in the Select Sires Inc. reproductive management program. Group 1 (high fertility bulls, n = 8) 1.9 ≤ PD ≤ 2.7%, and group 2 (low fertility bulls, n = 8) -6.5 ≤ PD ≤ 1.8 %. Additionally, the samples were categorized as processed (cryopreserved) or unprocessed (non-cryopreserved) for protein analysis. Protein expression was analyzed by 2-D fluorescence difference gel electrophoresis (2D-DIGETM). Protein spots were picked from a reference gel, analyzed by mass spectrometry and, subsequently identified by MS/MS ion searches performed on the SwissProt database. Protein expression did not differ (P > 0.05) with fertility grouping but displayed two distinct patterns among the processing groups: majority of the functional proteins were highly expressed in seminal plasma of non-cryopreserved semen while the cryopreserved semen contained mainly structural/extender derived proteins. Functional proteins identified included Spermadhesin Z13, BSP A1/2, BSP 30 kDa, Nucleobindin-1 and metalloproteinase inhibitor 2. Some of these proteins have been identified as anti-fertility or fertility enhancing agents in males. Whether this alteration in protein expression after processing might affect semen fertility is worthy of further evaluation.

Share and Cite:

Odhiambo, J. and Dailey, R. (2011) Characterization of proteins in cryopreserved and non-cryopreserved seminal plasma of dairy bulls of dif-fering fertility. Open Journal of Animal Sciences, 1, 33-40. doi: 10.4236/ojas.2011.12005.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Cancel, A.M., Chapman, D.A. and Killian, G.J. (1997) Osteopontin is the 55-kilodalton fertility-associated pro- tein in Holstein bull seminal plasma. Biology of Repro- duction, 57, 1293-1301.doi:10.1095/biolreprod57.6.1293
[2] Gwathmey, T.M., Ignotz, G.G. and Suarez, S.S. (2003) PDC-109 (BSP-A1/A2) promotes bull sperm binding to oviductal epithelium in vitro and may be involved in forming the oviductal sperm reservoir. Biology of Re- production, 69, 809-815. doi:10.1095/biolreprod.102.010827
[3] Gwathmey, T.M., Ignotz, G.G., Mueller, J.L., Manjunath, P. and Suarez S.S. (2006) Bovine seminal plasma pro-teins PDC-109, BSP-A3, and BSP-30-kDa share func-tional roles in storing sperm in the oviduct. Biology of Reproduction, 75, 501-507. doi:10.1095/biolreprod.106.053306
[4] Jobim, M.I., Oberst, E.R., Salbego, C.G., Souza, D.O., Wald, V.B., Tramontina, F. and Mattos, R.C. (2004) Two-dimensional polyacrylamide gel electrophoresis of bovine seminal plasma proteins and their relation with semen freezability. Theriogenology, 61, 255-266. doi:10.1016/S0093-691X(03)00230-9
[5] Killian, G.J., Chapman, D.A. and Rogowski, L.A. (1993) Fertility-associated proteins in Holstein bull seminal plasma. Biology of Reproduction, 49, 1202-1207. doi:10.1095/biolreprod49.6.1202
[6] Lowry, O.H., Rosebrough, N.J., Farr, A. L. and Randall, R.J. (1951) Protein measurement with the Folin-Phenol reagents. The Journal of Biological Chemistry, 193, 265-275.
[7] Manjunath, P., Bergeron, A., Lefebvre, J. and Fan, J. (2007) Seminal plasma proteins: Functions and interac- tion with protective agents during semen preservation. Society of Reproduction and Fertility Supplement, 65, 217-228.
[8] Mortarino, M., Tedeschi, G., Negri, A., Ceciliani, F., Got- tardi, L, Maffeo, G. and Ronchi, S. (1998) Two-dimen- sional polyacrylamide gel electrophoresis map of bull seminal plasma proteins. Electrophoresis, 19, 797-801. doi:10.1002/elps.1150190532
[9] Moura, A.A., Chapman, D.A. and Killian, G.J. (2007) Proteins of the accessory sex glands associated with the oocyte-penetrating capacity of cauda epididymal sperm from holstein bulls of documented fertility. Molecular Reproduction & Development, 74, 214-222. doi:10.1002/mrd.20590
[10] Moura, A.A., Koc H., Chapman D.A. and Killian G.J. (2006) Identification of proteins in the accessory sex gland fluid associated with fertility indexes of dairy bulls: a proteomic approach. Journal of Andrology, 27, 201-211. doi:10.2164/jandrol.05089
[11] O, W.S., Chen, H.Q. and Chow P.H. (1998) Effects of male accessory sex gland secretions on early embryonic development in the golden hamster. Journal of Repro- duction Fertiilityl, 84, 341-344.
[12] O’Leary, S., Robertson, S.A. and Armstrong, D.T. (2002) The influence of seminal plasma on ovarian function in pigs—a novel inflammatory mechanism? Journal of Re- productive Immunology, 57, 225-238.
[13] Odhiambo, J.F., Poole, D.H., Hughes, L., DeJarnette, M., Inskeep, E.K. and Dailey, R.A. (2009) Pregnancy out- come in dairy and beef cattle after artificial insemination and treatment with seminal plasma or transforming growth factor beta-1. Theriogenology, 72, 566-571. doi:10.1016/j.theriogenology.2009.04.013
[14] Robertson, S.A. (2007) Seminal fluid signaling in the female reproductive tract: Lessons from rodents and pigs. Journal of Animal Science, 85, E36-44. doi:10.2527/jas.2006-578
[15] Thérien, I., Moreau, R. and Manjunath, P. (1998) Major proteins of bovine seminal plasma and high-density lipo- protein induce cholesterol efflux from epididymal sperm. Biology of Reproduction, 59, 768-776. doi:10.1095/biolreprod59.4.768
[16] Thérien, I., Soubeyrand, S. and Manjunath, P. (1997) Major proteins of bovine seminal plasma modulate sperm capacitation by high-density lipoprotein. Biology of Re- production, 57, 1080-1088. doi:10.1095/biolreprod57.5.1080
[17] Yu, B., Zhao, Y., Zhao, W., Chen, F., Liu, Y., Zhang, J., Fu, W., Zong, Z., Yu, A. and Guan, Y. (2003) The inhibi- tory effect of BSP-A1/-A2 on protein kinase C and tyro- sine protein kinase. Cell Biochemistry and Function, 21, 183-188. doi:10.1002/cbf.1018

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.