Identification of a second follistatin gene in grass carp (Ctenopharyngodon idellus) and its regulatory function in myogenesis during embryogenesis

Follistatin can antagonize the function of myostatin as a competitive binding protein and promote muscle growth in vivo. Here, we report the isolation and characterization of a second follistatin gene fst2 in grass carp (Ctenopharyngodon idellus). The grass carp fst2 cDNA was 1,376 bp in length, with an open reading frame (ORF) encoding 350 amino acid residues. A relatively low sequence identity of 78% was found between grass carp Fst2 and its paralog Fst1. Sequence and phylogenetic analyses suggest that the grass carp fst2 originated from fish-specific gene duplication. In adult fish, fst2 mRNA expression was observed in most tissues but was strongly expressed in the eyes, muscles, skin and ovary. Grass carp fst2 mRNA could be detected as early as 16 h post-fertilization (hpf), while fst1 mRNA was detected throughout embryogenesis. Using in situ hybridization, fst2 transcripts were detected in the anterior somites at 24 hpf and in the brain and posterior somites at 36 hpf. Meanwhile, fst1 mRNA was transcribed mainly in the optic vesicle and at the cephalic mesoderm at 12 hpf, in the eyes, cephalic mesoderm and at the lateral edge of most somites at 24 hpf, and mainly in the brain at 36 hpf. Furthermore, overexpression of fst2 mRNA markedly affected the formation of the embryonic midline and somite structures. Based on comparisons with fst1, our findings suggest that fst2 retained the ancestral functions of regulating muscle development and growth during embryogenesis in grass carp.

► The duplicated follistatin gene fst2 was cloned from grass carp. ► Grass carp Fst2 and its paralog Fst1 have a relatively low sequence identity of 78%. ► Fst2 mRNA has a wide tissue distribution, especially eyes, muscles, skin and ovary. ► Fst2 and fst1 mRNA have different tissue and developmental expression patterns. ► Grass carp fst2 retained its ancestral regulatory functions during embryogenesis.