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Levels of BMP-6 mRNA in goat ovarian follicles and in vitro effects of BMP-6 on secondary follicle development

Published online by Cambridge University Press:  06 October 2011

Isana M.A. Frota ,
Cintia C.F. Leitão ,
José J.N. Costa ,
Robert van den Hurk ,
Márcia V.A. Saraiva ,
José R. Figueiredo  and
José R.V. Silva
Isana M.A. Frota
Affiliation:
Biotechnology Nucleus of Sobral – NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
Cintia C.F. Leitão
Affiliation:
Biotechnology Nucleus of Sobral – NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
José J.N. Costa
Affiliation:
Biotechnology Nucleus of Sobral – NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
Robert van den Hurk
Affiliation:
Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Márcia V.A. Saraiva
Affiliation:
Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceara, Fortaleza, CE, Brazil.
José R. Figueiredo
Affiliation:
Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceara, Fortaleza, CE, Brazil.
José R.V. Silva*
Affiliation:
Biotechnology Nucleus of Sobral – NUBIS, Federal University of Ceara, Rua Comandante Maurocélio Rocha Pontes 100, CEP 62042–280, Sobral, CE, Brazil.
*
All correspondence to: J.R.V. Silva. Biotechnology Nucleus of Sobral – NUBIS, Federal University of Ceara, Rua Comandante Maurocélio Rocha Pontes 100, CEP 62042–280, Sobral, CE, Brazil. Tel:/Fax: +55 88 36132603. e-mail: jrvsilva@ufc.br
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Summary

Expression of BMP-6 mRNA was quantified by real-time polymerase chain reaction (PCR) and the BMP-6 protein was demonstrated by immunohistochemistry in the primordial, primary, secondary, small and large antral follicles of goat. Furthermore, the influence of BMP-6 on increase in diameter, antrum formation and expression of BMP-6 and FSH-R in in vitro cultured secondary follicles was studied. Therefore, goat primordial, primary and secondary follicles, as well as small and large antral follicles were obtained and the mRNA levels of BMP-6 were quantified by PCR in real time. Expression of BMP-6 protein in goat follicles was demonstrated by immunohistochemistry. The influence of BMP-6 in the presence or absence of follicle-stimulating hormone (FSH) on both the development of secondary follicles and the expression of mRNA for BMP-6 and FSH-R was evaluated after 6 days of culture. Furthermore, the follicular diameter and the formation of the antrum were evaluated before and after 6 days of culture and compared by Kruskal–Wallis and chi-squared tests (P < 0.05), respectively. The results show that the level of mRNA for BMP-6 in primary and secondary follicles was significantly higher than in the primordial follicles (P < 0.05). Similar levels of BMP-6 mRNA were observed in cumulus–oocyte complexes and mural granulosa/theca cells from small and large antral follicles, respectively. BMP-6 protein was expressed in oocytes of all categories of follicles and in granulosa cells from secondary follicles onwards. Addition of BMP-6 to the culture medium increased the diameter of secondary follicles mainly by antrum formation after 6 days’ culture, in the presence or absence of FSH (P < 0.05). Furthermore, addition of FSH resulted in increased levels of BMP-6 mRNA in these follicles (P < 0.05). Simultaneous administration of FSH and BMP-6 enhanced the levels of FSH receptor (FSH-R) mRNA (P < 0.05). It is concluded that BMP-6 mRNA is increased during transition from primordial to primary/secondary follicles in the goat ovaries and that BMP-6 enhances the growth of cultured secondary follicles.

Keywords

BMP-6FSH-RGoatOvarian follicles
Type
Research Article
Information
Zygote , Volume 21 , Issue 3 , August 2013 , pp. 270 - 278
Copyright
Copyright © Cambridge University Press 2011 

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References

Abir, R., Ben-Haroush, A., Melamed, N., Felz, C., Krissi, H. & Fisch, B. (2008). Expression of bone morphogenetic proteins 4 and 7 and their receptors IA, IB, and II in human ovaries from fetuses and adults. Fertil. Steril. 89, 1430–40.CrossRefGoogle Scholar
Araújo, V.R., Magalhães, D.M., Silva, G.M., Chaves, R.N., Verde, I.B.L., Silva, C.M.G., Name, K.P.O., Báo, S.N., Campello, C.C., Silva, J.R.V., Tavares, L.M.T., Figueiredo, J.R. & Ribeiro, R.A.P. (2010). Bone morphogenetic protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro.Braz. J. Vet. Res. 30 (9), 770–81.Google Scholar
Campbell, B.K., Souza, C.J., Skinner, A.J., Webb, R. & Baird, D.T. (2006). Enhanced response of granulosa and theca cells from sheep carriers of the FecB mutation in vitro to gonadotropins and bone morphogenic protein-2, -4, and -6. Endocrinology 147, 1608–20.CrossRefGoogle ScholarPubMed
Elvin, J.A., Yan, C. & Matzuk, M.M. (2000). Oocyte-expressed TGF-β superfamily members in female fertility. Mol. Cell. Endocrinol. 159, 1–5.CrossRefGoogle ScholarPubMed
Erickson, G.F. & Shimasaki, S. (2003). The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle. Reprod. Biol. Endocrinol. 1, 1–20.CrossRefGoogle ScholarPubMed
Frota, I.M.A., Leitão, C.C.F., Costa, J.J.N., Brito, I.R., Van Den Hurk, R. & Silva, J.R.V. (2010). Stability of housekeeping genes and expression of locally produced growth factors and hormone receptors in goat preantral follicles. Zygote 19, 71–83.CrossRefGoogle ScholarPubMed
Glister, C., Kemp, C.F. & Knight, P.G. (2004). Bone morphogenetic protein (BMP) ligands and receptors in bovine ovarian follicle cells: actions of BMP-4, -6 and -7 on granulosa cells and differential modulation of Smad-1 phosphorylation by follistatin. Reproduction 127, 239–54.CrossRefGoogle ScholarPubMed
Glister, C., Satchell, L. & Knight, P.G. (2010). Changes in expression of bone morphogenetic proteins (BMPs), their receptors and inhibin co-receptor betaglycan during bovine antral follicle development: inhibin can antagonize the suppressive effect of BMPs on thecal androgen production. Reproduction 140, 699–12.CrossRefGoogle ScholarPubMed
Gutierrez, C.G., Ralph, J.H., Telfer, E.E, Wilmut, I & Webb, R. (2000). Growth and antrum formation of bovine preantral follicles in long-term culture in vitro. Biol. Reprod. 62, 1322–28.CrossRefGoogle ScholarPubMed
Hirshfield, A.N. (1991). Development of follicles in the mammalian ovarian. Internat. Rev. Cytol. 124, 43–101.CrossRefGoogle Scholar
Juengel, J.L., Reader, K.L., Bibby, A.H., Lun, S., Ross, I., Haydon, L.J. & McNatty, K.P. (2006). The role of bone morphogenetic proteins 2, 4, 6, and 7 during ovarian follicular development in sheep: contrast to rat. Reproduction 131, 501–13.CrossRefGoogle Scholar
Lucci, C.M., Amorim, C.A., Bao, S.N., Figueiredo, J.R., Rodrigues, A.P.R., Silva, J.R.V. & Goncalves, P.B.D. (1999). Effect of the interval of serial sections of ovarian tissue in the tissue chopper on the number of isolated caprine preantral follicles. Anim. Reprod. Sci. 56, 39–49.CrossRefGoogle ScholarPubMed
McConnell, N.A., Yunus, R.S., Gross, S.A., Bost, K.L., Clemens, M.G. & Hughes, F.M. (2002). Water permeability of an ovarian antral follicle is predominantly transcellular and mediated by aquaporins. Endocrinology 143, 2905–12.CrossRefGoogle ScholarPubMed
McNatty, K.P., Galloway, S.M., Wilson, T., Smith, P., Hudson, N.L. & O'Connell, A. (2005). Physiological effects of major genes affecting ovulation rate in sheep. Gen. Select. Evolut. 37, 25–38.CrossRefGoogle ScholarPubMed
Otsuka, F., Kelly, M. R. & Shimasaki, S. (2001). Biological function and cellular mechanisms of bone morphogenetic protein-6 in the ovary. J. Biol. Chem. 276, 32889–95.CrossRefGoogle ScholarPubMed
Rodgers, R.J. & Irving-Rodgers, H.F. (2010). Formation of the ovarian follicular antrum and follicular fluid. Biol. Reprod. 82, 1021–29.CrossRefGoogle ScholarPubMed
Saraiva, M.V.A., Celestino, J.J.H., Araújo, V.R., Chaves, R.N., Almeida, A.P., Lima-Verde, I.B., Duarte, A.B.G., Silva, G.M., Martins, F.S., Bruno, J.B., Matos, M.H.T., Campello, C.C., Silva, J.R.V. & Figueiredo, J.R. (2010). Expression of follicle-stimulating hormone receptor (FSHR) in goat ovarian follicles and the impact of sequential culture medium on in vitro development of caprine preantral follicles. Zygote 21, 1–10.Google Scholar
Shi, J., Yoshino, O., Osuga, Y., Koga, K., Hirota, Y., Hirata, T., Yano, T., Nishii, O. & Taketani, Y. (2009). Bone morphogenetic protein-6 stimulates gene expression of follicle-stimulating hormone receptor, inhibin/activin β subunits, and anti-Müllerian hormone in human granulosa cells. Fertil. Steril. 92, 1794–98.CrossRefGoogle ScholarPubMed
Shimasaki, S., Moore, R.K., Otsuka, F. & Erickson, G.F. (2004). The bone morphogenetic protein system in mammalian reproduction. Endocr. Rev. 25, 72–101.CrossRefGoogle ScholarPubMed
Silva, J.R.V., Van Den Hurk, R., Van Tol, H.T.A., Roelen, B.A.J. & Figueiredo, J.R. (2004a). Expression of growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15) and BMP receptors in the ovaries of goats. Mol. Reprod. Dev. 70, 11–19.CrossRefGoogle Scholar
Silva, J.R.V., Van den Hurk, R., Matos, M.H.T., Santos, R.R., Pessoa, C., Moraes, M.O. & Figueiredo, J.R. (2004b). Influences of FSH and EGF on primordial follicles during in vitro culture of caprine ovarian cortical tissue. Theriogenology 61, 1691–4.CrossRefGoogle ScholarPubMed
Souza, C.J.H., Campbell, B.K., McNeilly, A.S. & Baird, D.T. (2002). Effect of bone morphogenetic protein 2 (BMP2) on oestradiol and inhibin A production by sheep granulosa cells, and localization of BMP receptors in the ovary by immunohistochemistry. Reproduction 123, 363–69.CrossRefGoogle ScholarPubMed
Van den Hurk, R. & Zhao, J. (2005). Formation of mammalian oocytes and their growth, differentiation and maturation within ovarian follicles. Theriogenology 63, 1717–51.CrossRefGoogle ScholarPubMed
Van Tol, H.T.A., Van, E.I.J.K., Mummery, C.L., Van Den Hurk, R. & Bevers, M.M. (1996). Influence of FSH and hCG on the resumption of meiosis of bovine oocytes surrounded by cumulus cells connected to membrane granulosa. Mol. Reprod. Dev. 45, 218–24.3.0.CO;2-X>CrossRefGoogle ScholarPubMed
Xu, Y., Li, E., Han, Y., Chen, L. & Xie, Z. (2010). Differential expression of mRNAs encoding BMP/Smad pathway molecules in antral follicles of high- and low-fecundity Hu sheep. Anim. Reprod. Sci. 120 (1–4), 47–55.CrossRefGoogle ScholarPubMed