ABB  Vol.4 No.5 , May 2013
Expression of α6 integrin subunit in bovine oocyte and its potential role during fertilization
Abstract: Fertilization in mammals requires the successful completion of a sequence of steps, starting with the transport of gametes in the reproductive tract and ending with sperm-egg membrane fusion to produce a zygote. Although some integrin subunits are known to be associated with the plasma membrane of some mammalian oocytes and spermatozoa, the presence of α6 integrin on bovine oocytes with intact zona pellucida has not been reported. The present study was undertaken to evaluate the expression of α6 integrin subunit in bovine oocyte and to determine if in vitro binding to the zona pellucida and fertilization were affected by treating oocytes with α6 integrin subunit antibody. The α6 integrin subunit was identified on the bovine oocyte by immunocytochemistry. In vitro fertilization was significantly decreased when in vitro matured bovine oocytes were pre-incubated with α6 integrin subunit antibody at concentration 5 and 20 μg/mL, and spermoocyte binding increased. These studies demonstrated the presence of α6 integrin subunit on bovine oocyte, and its importance in fertilization and polyspermy.
Cite this paper: Gonçalves, R. , Bertolla, R. , Orlandi, C. and Barnabe, V. (2013) Expression of α6 integrin subunit in bovine oocyte and its potential role during fertilization. Advances in Bioscience and Biotechnology, 4, 617-622. doi: 10.4236/abb.2013.45081.

[1]   Gonçalves, R.F., Wolinetz, C.D. and Killian, G.J. (2007) Influence of arginine-glycine-aspartic acid (RGD), integrins (αV and α5) and osteopontin on bovine sperm-egg binding and fertilization in vitro. Theriogenology, 67, 468-474. doi:10.1016/j.theriogenology.2006.08.013

[2]   Tarone, G., Russo, M.A., Hirsch, E., Odorisio, T., Altruda, F., Silengo, L. and Siracusa, G. (1993) Expression of beta 1 integrin complexes on the surface of unfertilized mouse oocyte. Development, 117, 1369-1375.

[3]   Evans, J.P., Schultz, R.M. and Knopf, G.S. (1995) Identification and localization of integrin subunits in oocytes and eggs of the mouse. Molecular Reproduction and Development, 40, 220-221. doi:10.1002/mrd.1080400210

[4]   Bronson, R.A. and Fusi, F. (1990) Evidence that an arggly-asp adhesion sequence plays a role in mammalian fertilization. Biology of Reproduction, 43, 1019-1025. doi:10.1095/biolreprod43.6.1019

[5]   Bronson, R.A. and Fusi, F. (1990) Sperm-oolemmal interactions: Role of the Arg-Gly-Asp (RGD) adhesion peptide. Fertility and Sterility, 54, 527-529.

[6]   Zuccotti, M., Giorgi Rossi, P., Fiorillo, E., Garagna, S., Farabosco, A. and Redi, C.A. (1998) Timing of gene expression and oolemma localization of mouse alpha6 and beta1 integrin subunits during oogenesis. Developmental Biology, 200, 27-34. doi:10.1006/dbio.1998.8923

[7]   Burns, K.H., Owens, G.E., Fernandez, J.M., Nilson, J.H. and Matzuk, M.M. (2002) Characterization of integrin expression in the mouse ovary. Biology of Reproduction, 67, 743-751. doi:10.1095/biolreprod.101.000729

[8]   Boissonnas, C.C., Montjean, D., Lesaffre, C., Auer, J., Vaiman, D., Wolf, J.P. and Ziyyat, A (2010) Role of sperm alphavbeta3 integrin in mouse fertilization. Developmental Dynamics, 239, 773-783. doi:10.1002/dvdy.22206

[9]   White, K.L.; Pate, B.J. and Sessions, B.R. (2010) Oolema receptors and oocyte activation. Systems Biology in Reproductive Medicine, 56, 365-375.

[10]   Campbell, S., Swann, H.R. Seif, M.W., Kimber, S.J. and Aplin, J.D. (1995) Cell adhesion molecules on the oocyte and preimplantation human embryo. Human Reproduction, 10, 1572-1578. doi:10.1093/HUMREP/10.6.1571

[11]   Ji, Y.Z., Wolf, J.P., Jouannet, P. and Bomsel, M. (1998) Human gamete fusion can bypass beta 1 integrin requirement. Human Reproduction, 13, 682-689. doi:10.1093/humrep/13.3.682

[12]   Sengoku, K., Takuma, N., Miyamoto, T., Horikama, M. and Ishikawa, M. (2004) Integrins are not involved in the process of human sperm-oolemmal fusion. Human Reproduction, 19, 639-644. doi:10.1093/humrep/deh095

[13]   Ziyyat, A., Rubinstein, E., Monier-Gavelle, F., Barraud, V., Kulski, O., Prenant, M. Boucheix, C., Bomsel, M. and Wolf, J.P. (2006) CD9 controls the formation of clusters that contain tetraspaninas and the integrin alpha 6 beta 1, which are involved in human and mouse gamete fusion. Journal of Cell Science, 119, 416-424. doi:10.1242/jcs.02730

[14]   Reddy, V.R.K., Rajeev, S.K. and Gupta, V. (2003) Alpha 6 beta 1 integrin is a potential clinical marker for evaluating sperm quality in men. Fertility Sterility, 79, 15901600. doi:10.1016/S0015-0282(03)00368-6

[15]   Glander, H.J. and Schaller, J. (1993) Beta 1 integrins of spermatozoa: A flow cytophometric analysis. International Journal of Andrology, 16, 105-111. doi:10.1111/j.1365-2605.1993.tb01162.x

[16]   de Barros, F.R., Worst, R.A., Saurin, G.C., Mendes, C.M., Assumpção, M.E. and Visintin, J.A. (2012) α-6 integrin expression in bovine spermatogonial cells purified by discontinuous Percoll density gradient. Reproduction in Domestic Animals, 46, 887-890. doi:10.1111/j.1439-0531.2012.01985.x

[17]   Pate, B.J., White, K.L., Winger, Q.A., Rickords, L.F., Aston, K.L., Sessons, B.R., Li, G.P., Campbell, K.D., Weimer, B. and Bunch, T.D. (2007) Specific integrin subunits in bovine oocytes, including novel sequences for alpha 6 and beta 3 subunits. Molecular Reproduction and Development, 74, 600-607. doi:10.1002/mrd.20649

[18]   Primakoff, P. and Myles, D.G. (1983) A map of the guinea pig sperm surface constructed with monoclonal antibodies. Developmental Biology, 98, 417-428. doi:10.1016/0012-1606(83)90371-8

[19]   Allen, C.A. and Green, D.P. (1995) Monoclonal antibodies which recognize equatorial segmente epitops de novo following the A23187-indiced acrosome reaction of guinea pig sperm. Journal of Cell Science, 108, 767-777.

[20]   Toshimori, K., Saxena, D.K., Tanii, I. and Yoshinaga, K. (1998) An MN9 antigenic molecule, equatorial is required for successful sperm-oocyte fusion in mice. Biology of Reproduction, 59, 22-29. doi:10.1095/biolreprod59.1.22

[21]   Evans, J.P. (2012) Sperm-egg interaction. Annual Review of Physiology, 74, 447-502. doi:10.1146/annurev-physiol-020911-153339

[22]   Gonçalves, R.F., Barnabe, V.H. and Killian, G.J. (2008) Pre-treatment of cattle sperm and/or oocyte with antibody to lipocalin type prostaglandin D synthase inhibits in vitro fertilization and increases sperm-oocyte binding. Animal Reproduction Science, 106, 188-193. doi:10.1016/j.anireprosci.2007.12.019

[23]   Gonçalves, R.F., Staros, A.L. and Killian, G.J. (2008) Oviductal fluid proteins associated with the bovine zona pellucida and the effect on in vitro sperm-egg binding, fertilization and embryo development. Reproduction in Domestic Animals, 43, 720-729. doi:10.1111/j.1439-0531.2007.00978.x

[24]   Gonçalves, R.F., Wolinetz, C.G., Barnabe, V.H. and Killian, G.J. (2009) Influence of osteopontin in bovine uterine tube fluid on sperm binding and fertilization in RCA1 lectin-treated oocytes. Reproduction in Domestic Animals, 44, 152-155. doi:10.1111/j.1439-0531.2007.01011.x

[25]   Hynes, R.O. (1992) Integrins: Versatility, modulation, and signaling in cell adhesion. Cell, 69, 11-25. doi:10.1016/0092-8674(92)90115-S

[26]   Hazenbos, W.L., Van Den Berg, B.M. and Van Furth, R. (1993) Very late antigen-5 and complement receptor type 3 cooperatively mediate the interaction between Bordetella pertussis and human monocytes. Journal of Immunology, 151, 6274-6282.

[27]   Fénichel, P. and Durand-Clément, M. (1998) Role of integrins during fertilization in mammals. Human Reproduction, 13, 31-46. doi:10.1093/humrep/13.suppl_4.31

[28]   De Nadai, C., Fénichel, P., Donzeau, M., Epel, D. and Ciapa, B. (1996) Characterization and role of integrins during gametic interaction and egg activation. Zygote, 4, 3140. doi:10.1017/S0967199400002860

[29]   Almeida, E.A., Huovila, A.P., Sutherland, A.E., Stephens, L.E., Calarco, P.G., Shaw, L.M., Mercurio, A.M., Sonnenberg, A., Primakoff, P., Myles, D.G. and White, J.M. (1995) Mouse egg. Cell, 81, 1095-1104. doi:10.1016/S0092-8674(05)80014-5

[30]   Fusi, F.M., Vignale, M., Gailit, J. and Bronson, R.A. (1993) Mammalian oocytes exhibit specific recognition of the RGD (Arg-Gly-Asp) tripeptide and express oolemmal integrins. Molecular Reproduction and Development, 36, 212-219. doi:10.1002/mrd.1080360212

[31]   Campbell, K.D., Reed, W.A. and White, K.L. (2000) Ability of integrins to mediate fertilization, intracellular calcium release and parthenogenic development in bovine oocytes. Biology of Reproduction, 62, 1702-1709. doi:10.1095/biolreprod62.6.1702

[32]   Eto, K., Huet, C., Tarui, T., Kupriyanov, S., Liu, H.Z., Puzon-McLaughlin, W., Zhang, X.P., Sheppard, D., Engvall, E. and Takada, Y. (2002) Functional classification of ADAMs based on a conserved motif for binding to integrin alpha 9 beta 1: Implications for sperm-egg binding and other cell interactions. Journal of Biological Chemistry, 277, 17804-17810. doi:10.1074/jbc.M200086200

[33]   Gonçalves, R.F., Bertolla, R.P., Mortara, R.A. and Barnabe, V.H. (2008) Identification of some integrin subunits on cattle (Bos indicus) oocytes. Reproduction in Domestic Animals, 43, 147.

[34]   Pate, B.J., White, K.L., Winger, Q.A., Rickords, L.F., Aston, K.I. and Sessons, B.R. (2007) Specific integrin subunits in bovine oocytes, including novel sequences for alpha 6 and beta 3 subunits. Molecular Reproduction and Development, 74, 600-607. doi:10.1002/mrd.20649

[35]   Linfor, J. and Berger, T. (2000) Potential role of alphav and beta1 integrin as oocyte adhesion molecules during fertilization in pigs. Journal of Reproduction and Fertility, 120, 65-72.

[36]   Anderson, D.J., Abbot, A.F. and Jack, R.M. (1993) The role of complement C3b and its receptors in sperm-oocyte interaction. Proceedings of the National Academy of Science of the United States of America, 90, 1005110055. doi:10.1073/pnas.90.21.10051

[37]   Gardner, A.J. and Evans, J.P. (2006) Mammalian membrane block to polyspermy: New insights into how mammalian eggs prevent fertilization by multiple sperm. Reproduction Fertility and Development, 18, 53-61.