Growth differentiation factor 9 of Megalobrama amblycephala: molecular characterization and expression analysis during the development of early embryos and growing ovaries
- 327 Downloads
Growth differentiation factor 9 (GDF9) is a member of the transforming growth factorβ superfamily and plays an essential role during follicle maturation in mammals. In the present study, the full-length complementary DNA (cDNA) of gdf9 was obtained from Megalobrama amblycephala. The cDNA sequence is 2,061 bp in length with an open reading frame of 1,287 bp encoding 428 amino acid residues. The deduced amino acid sequence shared identities of about 42–86 % with the homologues of other vertebrates. During the early development of embryos, the gdf9 mRNA was detected in zygote with significantly high level and declined sharply by 47 and 87 % at 4 hours post-fertilization (hpf) and 6 hpf and even to an undetectable level through advancing stages. Expression analysis based on quantitative real-time PCR revealed that gdf9 mRNA was mainly expressed in ovary, but much lower levels were also found in some nonovarian tissues. Within the follicle, gdf9 mRNA was localized both in the oocytes and the follicle layer cells by in situ hybridization. During the ovarian cycle, gdf9 mRNA significantly decreased after the previtellogenic stage and became to increase again after the fully grown stage. The results imply that Gdf9 may play critical physiological functions in M. amblycephala early embryonic development and reproduction.
KeywordsMegalobrama amblycephala Growth differentiation factor 9 mRNA expression Early embryonic development Ovarian development
This research was supported by Program for New Century Excellent Talents in University (NCET-10-0403), and the Fundamental Research Funds for the Central Universities (2010PY004,2013PY067).
- Aaltonen J, Laitinen MP, Vuojolainen K, Jaatinen R, Horelli-Kuitunen N, Seppä L, Louhio H, Tuuri T, Sjöberg J, Bützow R, Hovata O, Dale L, Ritvos O (1999) Human growth differentiation factor 9 (GDF-9) and its novel homolog GDF-9B are expressed in oocytes during early folliculogenesis. J Clin Endocrinol Metab 84:2744–2750PubMedGoogle Scholar
- Eckery DC, Whale LJ, Lawrence SB, Wylde KA, McNatty KP, Juengel JL (2002) Expression of mRNA encoding growth differentiation factor 9 and bone morphogenetic protein 15 during follicular formation and growth in a marsupial, the brushtail possum (Trichosurus vulpecula). Mol Cell Endocrinol 192:115–126PubMedCrossRefGoogle Scholar
- García-López Á, Sánchez-Amaya MI, Halm S, Astola A, Prat F (2011) Bone morphogenetic protein 15 and growth differentiation factor 9 expression in the ovary of European sea bass (Dicentrarchus labrax): cellular localization, developmental profiles, and response to unilateral ovariectomy. Gen Comp Endocrinol 174:326–334PubMedCrossRefGoogle Scholar
- Halm S, Ibañez AJ, Tyler CR, Prat F (2008) Molecular characterisation of growth differentiation factor 9 (gdf9) and bone morphogenetic protein 15 (bmp15) and their patterns of gene expression during the ovarian reproductive cycle in the European sea bass. Mol Cell Endocrinol 291:95–103PubMedCrossRefGoogle Scholar
- Hanrahan JP, Gregan SM, Mulsant P, Mullen M, Davis GH, Powell R, Galloway SM (2004) Mutations in the genes for oocyte-derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare sheep (Ovis aries). Biol Reprod 70:900–909PubMedCrossRefGoogle Scholar
- Lokman PM, Kazeto Y, Ozaki Y, Ijiri S, Tosaka R, Kohara M, Divers SL, Matsubara H, Moore LG, Adachi S (2010) Effects of reproductive stage, GH, and 11-ketotestosterone on expression of growth differentiation factor-9 in the ovary of the eel, Anguilla australis. Reproduction 139:71–83PubMedCrossRefGoogle Scholar
- McGrath SA, Esquela AF, Lee SJ (1995) Oocyte-specific expression of growth/differentiation factor-9. Mol Endocrino 9:131–136Google Scholar
- Meйeн BA (1939) The ways of reproduction of migratory fishes. Trans Inst Marine Fish Oceanogr USSR 16:389–420Google Scholar
- Shimizu T, Miyahayashi Y, Yokoo M, Hoshino Y, Sasada H, Sato E (2004) Molecular cloning of porcine growth differentiation factor 9 (GDF-9) cDNA and its role in early folliculogenesis: direct ovarian injection of GDF-9 gene fragments promotes early folliculogenesis. Reproduction 128:537–543PubMedCrossRefGoogle Scholar
- Vitt UA, Hayashi M, Klein C, Hsueh AJ (2000b) Growth differentiation factor-9 stimulates proliferation but suppresses the follicle-stimulating hormone-induced differentiation of cultured granulosa cells from small antral and preovulatory rat follicles. Biol Reprod 62:370–377PubMedCrossRefGoogle Scholar