Skip to main content Accessibility help
×
Home

In vitro capacitation of bull spermatozoa by oviductal fluid and its components

  • Ann-Sofi Bergqvist (a1), Joan Ballester (a2), Anders Johannisson (a2), Marta Hernandez (a3), Nils Lundeheim (a4) and Heriberto Rodríguez-Martínez (a1)...

Summary

Sperm capacitation is crucial for fertilization. However, debate continues on exactly how, where and when capacitation is elicited in the bovine female genital tract. In this study we used merocyanine-540 and the chlortetracycline (CTC) assay to test how capacitation of bull spermatozoa is affected in vitro by exposure to oviductal fluid (ODF) collected in vivo, various glycosaminoglycans (GAGs) or bicarbonate. Following different durations of exposure, spermatozoa were stained with CTC or merocyanine-540, and evaluated with epifluorescent light microscopy or flow cytometry, respectively. Incubation time did not significantly affect capacitation. Exposure (30–120 min) to ODF capacitated (p < 0.05) bull spermatozoa as measured by either merocyanine-540 or CTC. Hyaluronan was the only GAG that induced a significant increase in B-pattern spermatozoa (capacitated; p = 0.012) compared with controls. Dermatan sulphate also induced capacitation (merocyanine-540 high fluorescence; p = 0.035). Exposure to bicarbonate-enriched media also yielded an increase in merocyanine-540 high fluorescence (p < 0.0001). When bicarbonate was added to the other treatments (ODF or GAGs) an equal increase in merocyanine-540 high fluorescence was noted (p < 0.0001), compared with before addition of bicarbonate and independent of the treatment before exposure. There was no significant difference in the number of B-pattern spermatozoa when bicarbonate was added, but an significant increase in spermatozoa with an acrosome-reacted (AR)-pattern (p < 0.0001) was observed. Exposure of spermatozoa to solubilized zonae pellucidae significantly increased the AR-pattern spermatozoa (p = 0.016). In conclusion, ODF was more potent in inducing capacitation of bull spermatozoa than the individual GAGs. Our results also indicate that bicarbonate is an effector of bull sperm capacitation.

Copyright

Corresponding author

All correspondence to: Ann-Sofi Bergqvist, DVM, Division of Comparative Reproduction, Obstetrics and Udder Health, Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences (SLU), PO Box 7054, 750 07 Uppsala, Sweden. Tel: +46 18 672174. Fax: +46 18 674535. e-mail: ann-sofi.bergqvist@kv.slu.se

References

Hide All
Abeydeera, L.R., Funahashi, H., Kim, N.H. & Day, B.N. (1997). Chlortetracycline fluorescence patterns and in vitro fertilisation of frozen-thawed boar spermatozoa incubated under various bicarbonate concentrations. Zygote 5, 117–25.
Ashworth, P.J., Harrison, R.A., Miller, N.G., Plummer, J.M. & Watson, P.F. (1995). Flow cytometric detection of bicarbonate-induced changes in lectin binding in boar and ram sperm populations. Mol. Reprod. Dev. 40, 164–76.
Bailey, J.L., Bilodeau, J.F. & Cormier, N. (2000). Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. J. Androl. 21, 17.
Bergqvist, A.S. & Rodriguez-Martinez, H. (2006). Sulphated glycosaminoglycans (S-GAGs) and syndecans in the bovine oviduct. Anim. Reprod. Sci. 93, 4660.
Bergqvist, A.S., Yokoo, M., Heldin, P., Frendin, J., Sato, E. & Rodríguez-Martínez, H. (2005). Hyaluronan and its binding proteins in the epithelium and intraluminal fluid of the bovine oviduct. Zygote 13, 207–18.
Cayli, S., Jakab, A., Ovari, L., Delpiano, E., Celik-Ozenci, C., Sakkas, D., Ward, D. & Huszar, G. (2003). Biochemical markers of sperm function: male fertility and sperm selection for ICSI. Reprod. Biomed. Online 7, 462–8.
Chamberland, A., Fournier, V., Tardif, S., Sirard, M.A., Sullivan, R. & Bailey, J.L. (2001). The effect of heparin on motility parameters and protein phosphorylation during bovine sperm capacitation. Theriogenology 55, 823–35.
Collin, S., Sirard, M.A., Dufour, M. & Bailey, J.L. (2000). Sperm calcium levels and chlortetracycline fluorescence patterns are related to the in vivo fertility of cryopreserved bovine semen. J. Androl. 21, 938–43.
Cormier, N., Sirard, M.A. & Bailey, J.L. (1997). Premature capacitation of bovine spermatozoa is initiated by cryopreservation. J. Androl. 18, 461–68.
DasGupta, S., Mills, C.L. & Fraser, L.R. (1993). Ca2+-related changes in the capacitation state of human spermatozoa assessed by a chlortetracycline fluorescence assay. J. Reprod. Fertil. 99, 135–43.
Flesch, F.M. & Gadella, B.M. (2000). Dynamics of the mammalian sperm plasma membrane in the process of fertilization. Biochim. Biophys. Acta 10, 197235.
Flesch, F.M., Brouwers, J.F., Nievelstein, P.F., Verkleij, A.J., van Golde, L.M., Colenbrander, B. & Gadella, B.M. (2001). Bicarbonate stimulated phospholipid scrambling induces cholesterol redistribution and enables cholesterol depletion in the sperm plasma membrane. J. Cell Sci. 114, 3543–55.
Florman, H.M. & First, N.L. (1988). The regulation of acrosomal exocytosis. Sperm capacitation is required for the induction of acrosome reactions by the bovine zona pellucida in vitro. Dev. Biol. 128, 453–63.
Fraser, L.R., Abeydeera, L.R. & Niwa, K. (1995). Ca2+-regulating mechanisms that modulate bull sperm capacitation and acrosomal exocytosis as determined by chlortetracycline analysis. Mol. Reprod. Dev. 40, 233–41.
Gil, J., Januskauskas, A., Håård, M.Ch., Håård, M.G.M., Johanisson, A., Söderquist, L. & Rodriguez-Martinez, H. (2000). Functional sperm parameters and fertility of bull semen extended in biociphos Plus and Triladyl. Reprod. Dom. Anim. 35, 6977.
Grippo, A.A., Way, A.L. & Killian, G.J. (1995). Effect of bovine ampullary and isthmic oviductal fluid on motility, acrosome reaction and fertility of bull spermatozoa. J. Reprod. Fertil. 105, 5764.
Guthrie, H.D. & Welch, G.R. (2005). Impact of storage prior to cryopreservation on plasma membrane function and fertility of boar sperm. Theriogenology 15, 396410.
Handrow, R.R., Lenz, R.W. & Ax, R.L. (1982). Structural comparisons among glycosaminoglycans to promote an acrosome reaction in bovine spermatozoa. Biochem. Biophys. Res. Commun. 107, 1326–32.
Harrison, R.A. & Gadella, B.M. (2005). Bicarbonate-induced membrane processing in sperm capacitation. Theriogenology 15, 342–51.
Harrison, R.A., Ashworth, P.J. & Miller, N.G. (1996). Bicarbonate/CO2, an effector of capacitation, induces a rapid and reversible change in the lipid architecture of boar sperm plasma membranes. Mol. Reprod. Dev. 45, 378–91.
Hunter, R.H.F. & Rodriguez-Martinez, H. (2004). Capacitation of mammalian spermatozoa in vivo, with a specific focus on events in the fallopian tubes. Mol. Reprod. Dev. 67, 243–50.
Huszar, G., Willetts, M. & Corrales, M. (1990). Hyaluronic acid (Sperm Select) improves retention of sperm motility and velocity in normospermic and oligospermic specimens. Fertil. Steril. 54, 1127–34.
Januskaukas, A., Gil, J., Söderquist, L. & Rodriguez-Martinez, H. (2000). Relationship between sperm response to glycosaminoglycans in vitro and non-return rates in bull Swedish dairy AI bulls. Reprod. Dom. Anim. 35, 207–12.
Kawakami, E., Hori, T. & Tsutsui, T. (1998). Induction of dog sperm capacitation by oviductal fluid. J. Vet. Med. Sci. 60, 197202.
Kawakami, E., Arai, T., Oishi, I., Hori, T. & Tsutsui, T. (2000). Induction of dog sperm capacitation by glycosaminoglycans and glycosaminoglycan amounts of oviductal fluid and uterine fluid in bitches. J. Vet. Med. Sci. 62, 65–8.
Keskintepe, L. & Brackett, B.G. (1996). In vitro developmental competence of in vitro-matured bovine oocytes fertilized and cultured in completely defined media. Biol. Reprod. 55, 333–9.
Kjellen, L. & Lindahl, U. (1991). Proteoglycans: structure and interactions. Annu. Rev. Biochem. 60, 443–75.
Lee, C.N. & Ax, R.L. (1996). Concentrations and composition of glycosaminoglycans in the female bovine reproductive tract. J. Dairy Sci. 67, 2006–9.
Lee, C.N., Clayton, M.K., Bushmeyer, S.M., First, N.L. & Ax, R.L. (1986). Glycosaminoglycans in ewe reproductive tracts and their influence on acrosome reactions in bovine spermatozoa in vitro. J. Anim. Sci. 63, 861–7.
Lee, M.A. & Storey, B.T. (1986). Bicarbonate is essential for fertilization of mouse eggs: mouse sperm require it to undergo the acrosome reaction. Biol. Reprod. 34, 349–56.
Lefebvre, R. & Suarez, S.S. (1996). Effect of capacitation on bull sperm binding to homologous oviductal epithelium. Biol. Reprod. 54, 575–82.
Lefebvre, R., Chenoweth, P.J., Drost, M., LeClear, C.T., MacCubbin, M., Dutton, J.T. & Suarez, S.S. (1995). Characterization of the oviductal sperm reservoir in cattle. Biol. Reprod. 53, 1066–74.
Libersky, E.A. & Boatman, D.E. (1995). Progesterone concentrations in serum, follicular fluid, and oviductal fluid of the golden hamster during the preovulatory period. Biol. Reprod. 53, 477–82.
Mahmoud, A.I. & Parrish, J.J. (1996). Oviduct fluid and heparin induce similar surface changes in bovine sperm during capacitation: a flow cytometric study using lectins. Mol. Reprod. Dev. 43, 554–60.
Maxwell, W.M. & Johnson, L.A. (1997). Chlortetracycline analysis of boar spermatozoa after incubation, flow cytometric sorting, cooling, or cryopreservation. Mol. Reprod. Dev. 46, 408–18.
McNutt, T.L. & Killian, G.J. (1991). Influence of bovine follicular and oviduct fluids on sperm capacitation in vitro. J. Androl. 12, 244–52.
Meizel, S. & Turner, K.O. (1986). Glycosaminoglycans stimulate the acrosome reaction of previously capacitated hamster sperm. J. Exp. Zool. 237, 137–9.
Miller, D.J. & Hunter, A.G. (1986). Effect of osmolality and glycosaminoglycans on motility, capacitation, acrosome reaction, and in vitro fertilizability of bovine ejaculated sperm. J. Dairy Sci. 69, 2915–24.
Nagy, S., Jansen, J., Topper, E.K. & Gadella, B.M. (2003). A triple-stain flow cytometric method to assess plasma- and acrosome-membrane integrity of cryopreserved bovine sperm immediately after thawing in presence of egg-yolk particles. Biol. Reprod. 68, 1828–35.
Parrish, J.J., Susko-Parrish, J.L., Winer, M.A. & First, N.L. (1988). Capacitation of bovine sperm by heparin. Biol. Reprod. 38, 1171–80.
Parrish, J.J., Susko-Parrish, J.L., Handrow, R.R., Sims, M.M. & First, M.L. (1989 a). Capacitation of bovine spermatozoa by oviduct fluid. Biol. Reprod. 40, 1020–5.
Parrish, J.J., Susko-Parrish, J.L., Handrow, R.R., Ax, R.L. & First, N.L. (1989 b). Effect of sulfated glycoconjugates on capacitation and the acrosome reaction of bovine and hamster spermatozoa. Gamete Res. 24, 403–13.
Parrish, J.J., Susko-Parrish, J.L., Uguz, C. & First, N.L. (1994). Differences in the role of cyclic adenosine 3',5'-monophosphate during capacitation of bovine sperm by heparin. Biol. Reprod. 51, 1099–108.
Peña, A.I., Johannisson, A. & Linde-Forsberg, C. (1999). Post-thaw evaluation of dog spermatozoa using new triple fluorescent staining and flow cytometry. Theriogenology 15, 965–80.
Pollard, J.W., Plante, C., King, W.A., Hansen, P.J., Betteridge, K.J. & Suarez, S.S. (1991). Fertilizing capacity of bovine sperm may be maintained by binding of oviductal epithelial cells. Biol. Reprod. 44, 102–7.
Rathi, R., Colenbrander, B., Bevers, M.M. & Gadella, B.M. (2001). Evaluation of in vitro capacitation of stallion spermatozoa. Biol. Reprod. 65, 462–70.
Rodriguez-Martinez, H. (2001). Oviduct function in cows and pigs: with special reference to sperm capacitation. Asian-Aust. J. Anim. Sci. 14, 2837.
Rodríguez-Martínez, H., Han, Y., Song, X., Funahashi, H. & Niwa, K. (1997). Additive effects of sodium hyaluronate as inducer of capacitation of boar spermatozoa in vitro. Proceedings of the 5th International Conference on Pig Reproduction, June 2–4, The Netherlands.
Rodriguez-Martinez, H., Tienthai, P., Suzuki, K., Funahashi, H., Ekwall, H. & Johannisson, A. (2001). Involvement of oviduct in sperm capacitation and oocyte development in pigs. Reproduction Suppl. 58, 129–45.
Rodriguez-Martinez, H., Saravia, F., Wallgren, M., Tienthai, P., Johannisson, A., Vazquez, J.M., Martinez, E., Roca, J., Sanz, L. & Calvete, J.J. (2005). Boar spermatozoa in the oviduct. Theriogenology 15, 514–35.
Setchell, B.P. (1994). Possible physiological bases for contraceptive techniques in the male. Hum. Reprod. 9, 1081–7.
Shamsuddin, M., Rodriguez-Martinez, H. & Larsson, B. (1993). Fertilizing capacity of bovine spermatozoa selected after swim-up in hyaluronic acid-containing medium. Reprod. Fertil. Dev. 5, 307–15.
Sostaric, E., van de Lest, C.H., Colenbrander, B. & Gadella, B.M. (2005). Dynamics of carbohydrate affinities at the cell surface of capacitating bovine sperm cells. Biol. Reprod. 72, 346–57.
Suarez, S.S. (2002). Formation of a reservoir of sperm in the oviduct. Reprod. Domest. Anim. 37, 140–3.
Sugahara, K., Mikami, T., Uyama, T., Mizuguchi, S., Nomura, K. & Kitagawa, H. (2003). Recent advances in the structural biology of chondroitin sulfate and dermatan sulfate. Curr. Opin. Struct. Biol. 13, 612–20.
Suzuki, K., Asano, A., Eriksson, B., Niwa, K., Nagai, T. & Rodriguez-Martinez, H. (2002). Capacitation status and in vitro fertility of boar spermatozoa: effects of seminal plasma, cumulus-oocyte-complexes-conditioned medium and hyaluronan. Int. J. Androl. 25, 8493.
Tajik, P., Wang, W., Okuda, K. & Niwa, K. (1994). In vitro fertilization of bovine oocytes in a chemically defined, protein-free medium varying the bicarbonate concentration. Biol. Reprod. 50, 1231–7.
Therien, I., Bergeron, A., Bousquet, D. & Manjunath, P. (2005). Isolation and characterization of glycosaminoglycans from bovine follicular fluid and their effect on sperm capacitation. Mol. Reprod. Dev. 71, 97106.
Thundathil, J., Gil, J., Januskauskas, A., Larsson, B., Soderquist, L., Mapletoft, R. & Rodriguez-Martinez, H. (1999). Relationship between the proportion of capacitated spermatozoa present in frozen-thawed bull semen and fertility with artificial insemination. Int. J. Androl. 22, 366–73.
Tienthai, P., Johannisson, A. & Rodriguez-Martinez, H. (2004). Sperm capacitation in the porcine oviduct. Anim. Reprod. Sci. 80, 131–46.
Ward, C.R. & Storey, B.T. (1984). Determination of the time course of capacitation in mouse spermatozoa using a chlortetracycline fluorescence assay. Dev. Biol. 104, 287–96.
Way, A.L. & Killian, G.J. (2002). Capacitation and induction of the acrosome reaction in bull spermatozoa with norepinephrine. J. Androl. 23, 352–7.
Visconti, P.E., Westbrook, V.A., Chertihin, O., Demarco, I., Sleight, S. & Diekman, A.B. (2002). Novel signalling pathways involved in sperm acquisition of fertilizing capacity. J. Reprod. Immunol. 53, 133–50.
Yanagimachi, R. (1989). Sperm capacitation and gamete interaction. J. Reprod. Fertil. 38, 2733.
Yanagimachi, R. (1994). Fertility of mammalian spermatozoa: its development and relativity. Zygote 2, 371–2.

Keywords

In vitro capacitation of bull spermatozoa by oviductal fluid and its components

  • Ann-Sofi Bergqvist (a1), Joan Ballester (a2), Anders Johannisson (a2), Marta Hernandez (a3), Nils Lundeheim (a4) and Heriberto Rodríguez-Martínez (a1)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed