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

Zygote. 2013 Aug;21(3):270-8. doi: 10.1017/S0967199411000542. Epub 2011 Oct 6.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 6 / genetics*
  • Bone Morphogenetic Protein 6 / metabolism
  • Bone Morphogenetic Protein 6 / pharmacology
  • Culture Media / chemistry
  • Culture Media / pharmacology
  • Cumulus Cells / metabolism
  • Female
  • Follicle Stimulating Hormone / metabolism
  • Follicle Stimulating Hormone / pharmacology
  • Gene Expression Regulation, Developmental
  • Granulosa Cells / drug effects
  • Granulosa Cells / physiology
  • Ovarian Follicle / drug effects
  • Ovarian Follicle / growth & development
  • Ovarian Follicle / physiology*
  • Ovary / physiology
  • RNA, Messenger / metabolism
  • Receptors, FSH / genetics
  • Tissue Culture Techniques

Substances

  • Bone Morphogenetic Protein 6
  • Culture Media
  • RNA, Messenger
  • Receptors, FSH
  • Follicle Stimulating Hormone