RESUMO
Growth and differentiation factor-11 (GDF-11) is a member of the transforming growth factor-ß superfamily and is thought to be derived together with myostatin (known also as GDF-8) from an ancestral gene. In the present study, we report the isolation and characterization of GDF-11 homolog from a marine teleost, the gilthead sea bream Sparus aurata, and show that this growth factor is highly conserved throughout vertebrates. Using bioinformatics, we identified GDF-11 in Tetraodon, Takifugu, medaka, and stickleback and found that they are highly conserved at the amino acid sequence as well as gene organization. Moreover, we found conservation of syntenic relationships among vertebrates in the GDF-11 locus. Transcripts for GDF-11 can be found in eggs and early embryos, albeit at low levels, while in post-hatching larvae expression levels are high and decreases as development progresses, suggesting that GDF-11 might have a role during early development of fish as found in tetrapods and zebrafish. Finally, GDF-11 is expressed in various tissues in the adult fish including muscle, brain, and eye.
Assuntos
Sequência Conservada , Fatores de Diferenciação de Crescimento/química , Fatores de Diferenciação de Crescimento/genética , Dourada/genética , Homologia de Sequência de Aminoácidos , Sequência de Aminoácidos , Animais , Sequência de Bases , Evolução Biológica , Mapeamento Cromossômico , Biologia Computacional , Bases de Dados Genéticas , Peixes/genética , Fatores de Diferenciação de Crescimento/metabolismo , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Dourada/classificação , Dourada/crescimento & desenvolvimento , Dourada/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Smegmamorpha/genética , SinteniaRESUMO
BACKGROUND: The transforming growth factor-beta (TGF-beta) family constitutes of dimeric proteins that regulate the growth, differentiation and metabolism of many cell types, including that of skeletal muscle in mammals. The potential role of TGF-betas in fish muscle growth is not known. RESULTS: Here we report the molecular characterization, developmental and tissue expression and regulation by nutritional state of a novel TGF-beta gene from a marine fish, the gilthead sea bream Sparus aurata. S. aurata TGF-beta6 is encoded by seven exons 361, 164, 133, 111, 181, 154, and 156 bp in length and is translated into a 420-amino acid peptide. The exons are separated by six introns: >643, 415, 93, 1250, 425 and >287 bp in length. Although the gene organization is most similar to mouse and chicken TGF-beta2, the deduced amino acid sequence represents a novel TGF-beta that is unique to fish that we have named TGF-beta6. The molecule has conserved putative functional residues, including a cleavage motif (RXXR) and nine cysteine residues that are characteristic of TGF-beta. Semi-quantitative analysis of TGF-beta6 expression revealed differential expression in various tissues of adult fish with high levels in skin and muscle, very low levels in liver, and moderate levels in other tissues including brain, eye and pituitary. TGF-beta6 is expressed in larvae on day of hatching and increases as development progresses. A fasting period of five days of juvenile fish resulted in increased levels of TGF-beta6 expression in white skeletal muscle compared to that in fed fish, which was slightly attenuated by one injection of growth hormone. CONCLUSION: Our findings provide valuable insights about genomic information and nutritional regulation of TGF-beta6 which will aid the further investigation of the S. aurata TGF-beta6 gene in association with muscle growth. The finding of a novel TGF-beta6 molecule, unique to fish, will contribute to the understanding of the evolution of the TGF-beta family of cytokines in vertebrates.
