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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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