Macedonian Veterinary Review

Original Scientific Article

Effect of seminal plasma protein fractions on cooled dog semen kinetics

Tsvetan Stefanov Tsvetkov *

Denica Boyanova Daskalova

Mac Vet Rev 2023; 46 (2): 177 - 183

10.2478/macvetrev-2023-0027

Received: 28 November 2022

Received in revised form: 08 September 2023

Accepted: 13 September 2023

Available Online First: 22 September 2023

Published on: 15 October 2023

Correspondence: Tsvetan Stefanov Tsvetkov, tsvetan_tsvetkov_88@abv.bg

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Abstract

Semen cooling and cryopreservation have revolutionized the field of reproductive biotechnologies. However, challenges persist in maintaining sperm quality and viability during these processes. The unsatisfactory results and the main problems are associated with low quality, viability, morphological, structural, and DNA integrity, changes in plasma membrane, ability to interact with female tract and decrease in sperm fertilization potential. The objective of this study was to determine the effect of seminal plasma proteins with different molecular weights on the kinetic parameters of dog spermatozoa during cold storage at 4 °C. The proteins were isolated from the dogs’ seminal plasma ranging between 6-200 kDa, separated in four fractions. The ejaculates (n=15) were collected from 6 heathy dogs which were pooled. Spermatozoa were extended with Tris medium alone (control) or with addition of one of the isolated protein fractions, and were then incubated at 4 °C for 2 hours. Sperm incubated with seminal plasma proteins ranging between 10-15 kDa had significantly higher total motility (37.01±2.98%), sperm progressiveness (15.97±1.91%), curvilinear line velocity (37.46±3.75 μm/s), linearity (26.18±1.00%), and straightness (45.94±2.03%) compared to the other groups incubated with higher molecular weight proteins and the control group. The findings of this study indicated that the use of certain proteins in seminal plasma can be beneficial for reducing the detrimental effect of cooling at 4 °C and preserving the viability of dog spermatozoa. It seemed that the presence of 10-15 kDa proteins from canine seminal plasma rendered the spermatozoa less amenable to the negative influence during cooling.

Keywords: spermatozoa, seminal plasma proteins, cooling, kinetics, dog

References

1. England, G.C.W. (1993). Cryopreservation of dog semen: a review. J Reprod Fertil Suppl. 47, 243-255.

2. Martínez-Barbitta, M., Rivera Salinas, C. (2022). Evaluation of chilled dog semen extended with sperm activator. Front Vet Sci. 8, 764750. https://doi.org/10.3389/fvets.2021.764750 PMid:35224070 PMCid:PMC8874018

3. Michael, J., Alexopoulos, C., Pontiki, A., Hadjipavlou-Litina, D., Saratsis, P., Ververidis, N., Boscos, C. (2010). Effect of N-acetyl-L-cysteine supplementation in semen extenders on semen quality and reactive oxygen species of chilled canine spermatozoa. Reprod Domest Anim. 45(2): 201-207. https://doi.org/10.1111/j.1439-0531.2008.01202.x PMid:19508647

4. Nguyen, V., Ponchunchoovong, S., Kupittayanant, S., Kupittayanant, P. (2019). Effects of egg yolk and soybean lecithin on sperm quality determined by computer-assisted sperm analysis and confocal laser scanning microscope in chilled canine sperm. Vet Med Sci. 5(3): 345-360. https://doi.org/10.1002/vms3.158 PMid:30848107 PMCid:PMC6682803

5. Leahy, T., Gadella, M. (2011). Capacitation and capacitation-like sperm surface changes induced by handling boar semen. Reprod Domest Anim. 2, 7-13. https://doi.org/10.1111/j.1439-0531.2011.01799.x PMid:21884270
6. Caballero, I., Vazquez, J., Garcia, M., Parrilla, I., Roca, J., Calvete, J., Sanz, L., Martinez, A (2008). Major proteins of boar seminal plasma as a tool for biotechnological preservation of spermatozoa. Theriogenology 70(8): 1352-1355. https://doi.org/10.1016/j.theriogenology.2008.07.013 PMid:18771797
7. McKinnon, A. (1999). Breeding its technology - now the future. c2019 [cited 2019 December 22]. https://www.harness.org.au/hra/papers/PMCKINN5.HTM
8. Fuller, J., Whittingham, D. (1997). Capacitation-like changes occur in mouse spermatozoa cooled to low temperatures. Mol Reprod Dev. 46(3): 318-324. https://doi.org/10.1002/(SICI)1098-2795(199703)46:3<318::AID-MRD10>3.0.CO;2-V
9. Andrade, A.F.C., Knox, R.V., Torres, M.A., Pavaneli, A.P. (2022). What is the relevance of seminal plasma from a functional and preservation perspective? Anim Reprod Sci. 246, 106946. https://doi.org/10.1016/j.anireprosci.2022.106946 PMid:35216848
10. Bubenickova, F., Postlerova, P., Simonik, O., Sirohi, J., Sichtar, J. (2020). Effect of seminal plasma protein fractions on stallion sperm cryopreservation. Int J Mol Sci. 21(17): 6415. https://doi.org/10.3390/ijms21176415 PMid:32899253 PMCid:PMC7504567
11. Pace, M.M., Graham, E.F. (1974). Components in egg yolk which protect bovine sperm during freezing. Anim Sci J. 39(6): 1144-1149. https://doi.org/10.2527/jas1974.3961144x PMid:4475048
12. Mogielnicka-Brzozowska, M., Dziekońska, A., Strzeżek, R., Załęcki, M., Majewska, M., Tołścik, K., Kordan, W. (2014). Effect of seminal plasma zincbinding proteins on motility and membrane integrity of canine spermatozoa stored at 5°C. Bull Vet Inst Pulawy. 58, 163-168. https://doi.org/10.2478/bvip-2014-0025
13. Muiño-Blanco, T., Pérez-Pé, R., Cebrián-Pérez, J. (2008). Seminal plasma proteins and sperm resistance to stress. Reprod Domest Anim. Suppl 4, 18-31. https://doi.org/10.1111/j.1439-0531.2008.01228.x PMid:18803753
14. Pérez-Pé, R., Cebrián-Pérez, J.A., MuiñoBlanco, T. (2001). Semen plasma proteins prevent coldshock membrane damage to ram spermatozoa. Theriogenology 56(3): 425-434. https://doi.org/10.1016/S0093-691X(01)00574-X PMid:11516122
15. Tsutsui, T., Hase, M., Hori, T., Ito,T., Kawakami, E. (2000). Effects of orvus ES paste on canine spermatozoal longevity after freezing and thawing. J Vet Med Sci. 62(5): 533-535. https://doi.org/10.1292/jvms.62.533 PMid:10852405
16. Mogielnicka-Brzozowska, M., Fraser, L., Czarzasta, J., Kordan, W. (2012). Isolation and characterization of zinc-binding proteins of canine seminal plasma. Pol J Vet Sci. 15(3): 493-498. https://doi.org/10.2478/v10181-012-0076-5 PMid:23214370
17. Mogielnicka-Brzozowska, M., Wysocki, P., Strzezek, J., Kordan, W. (2011). Zinc-binding proteins from boar seminal plasma - isolation, biochemical characteristics and influence on spermatozoa stored at 4°C. Acta Biochim Pol. 58(2): 171-177. https://doi.org/10.18388/abp.2011_2261 PMid:21584285
18. Boursnell, J.C., Noble, E.A. (1975). The absorption by boar spermatozoa of zinc bound to low molecular weight ligands. J Reprod Fertil. 45(3): 421-426. https://doi.org/10.1530/jrf.0.0450421 PMid:1206641
19. Henkel, R., Bittner, J., Weber, M.D.R., Huther, F., Miska, W. (1999). Relevance of zinc in human sperm flagella and its relation to motility. Fertil Steril. 71(6): 1138-1143. https://doi.org/10.1016/S0015-0282(99)00141-7 PMid:10360924
20. Bjorndhal, L., Kvist, U. (1990). Influence of seminal vesicular fluid on the zinc content of human sperm chromatin. Int J Androl. 13(3): 232-237. https://doi.org/10.1111/j.1365-2605.1990.tb00981.x PMid:2387643
21. Nothling, O., Gerber, D., Colenbrander, B., Dijkstra, M., Bakker, T., De Cramer, K. (2007). The effect of homologous prostatic fluid on motility and morphology of dog epididymal spermatozoa extended and frozen in Biladyl with Equex STM paste or Andromed. Theriogenology 67(2): 264-275. https://doi.org/10.1016/j.theriogenology.2006.07.018 PMid:16962165
22. O’Meara, C., Donovan, A., Hanrahan, J., Duffy, P., Fair, S., Evans, A., Lonergan, P. (2007). Resuspending ram spermatozoa in seminal plasma after cryopreservation does not improve pregnancy rate in cervically inseminated ewes. Theriogenology 67(7): 1262-1268. https://doi.org/10.1016/j.theriogenology.2007.01.012 PMid:17336375
23. Pan, C., Wu, Y., Yang, Q., Yem, J. (2018). Effects of seminal plasma concentration on sperm motility and plasma and acrosome membrane integrity in chilled canine spermatozoa. Pol J Vet Sci. 21(1): 133-138.
24. Rota, A., Milani, C., Romagnoli, S. (2007). Effect of post-thaw dilution with autologous prostatic fluid on dog semen motility and sperm acrosome status. Theriogenology 67(3): 520-525. https://doi.org/10.1016/j.theriogenology.2006.09.001 PMid:17010416
25. Leahy, T., De Graaf, S. (2012). Seminal plasma and its effect on ruminant spermatozoa during processing. Reprod Domestic Anim. 47(Suppl. 4): 207-213. https://doi.org/10.1111/j.1439-0531.2012.02077.x PMid:22827372
26. Rueda, F., Quím, A., Garcés, L., Herrera, P., Arbeláez, L., Peña, R., Velásquez, J., Hernández Cardozo, C. (2013). Seminal plasma proteins increase the postthaw sperm viability of Sanmartinero bull’s semen. Rev MVZ Cordoba. 18(1): 3327-3335. https://doi.org/10.21897/rmvz.195
27. Caballero, I., Vazquez, M., Garcia, M., Parrilla, I., Roca, J., Calvete, J., Sanz, L., Martinez, A. (2008). Major proteins of boar seminal plasma as a tool for biotechnological preservation of spermatozoa. Theriogenology 70(8): 1352-1355. https://doi.org/10.1016/j.theriogenology.2008.07.013 PMid:18771797
28. Barrios, B., Pérez-Pé, R., Gallego, M., Tato, A., Osada, J., Muiño-Blanco, T., Cebrián-Pérez J. (2000). Seminal plasma proteins revert the coldshock damage on ram Sperm membrane. Biol Reprod. 63(5): 1531-1537. https://doi.org/10.1095/biolreprod63.5.1531 PMid:11058562
29. Baas, J.W., Molan, P.C., Shannon, P. (1983). Factors in seminal plasma of bulls that affect the viability and motility of spermatozoa. J Reprod Fertil. 68(2): 275-280. https://doi.org/10.1530/jrf.0.0680275 PMid:6864642
30. Amann, R.P., Cristanelli, M.J., Squires, E.L. (1987). Proteins in stallion seminal plasma. J Reprod Fertil. Suppl. 35, 113-120.

Copyright

© 2023 Stefanov Tsvetkov Ts. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Conflict of Interest Statement

The authors declared that they have no potential lict of interest with respect to the authorship and/or publication of this article.

Citation Information

Macedonian Veterinary Review. Volume 46, Issue 2, Pages 177-183, e-ISSN 1857-7415, p-ISSN 1409-7621, DOI: 10.2478/macvetrev-2023-0027