Study Of Expression Of Follistatin In Flounder (Paralichthys Olivaceus) Embryos And The Function On Development Of Muscle

Follistatin (FS) was first isolated from ovarian follicular fluid of cow and pig by Robertson and Ueno in 1987 respectively. It was a cysteine-rich single chain peptide. Follistatin has broad function on different type of cells. In this study, genomic sequence of Flounder Follistatin gene was identified from genome library of flounder (Paralichthys olivaceus) by Genomic Walking. The expression of the gene in embryo and adult tissues was analyzed by RT-PCR and in situ hybridization. The tissue-specific expression of the promoter was analyzed.1. The genomic sequence of flounder Follistatin was 4.3 kb long. The promoter was about 1.0 kb long. Compared with cDNA sequence revealed that there were five exons and four introns and the boundary of exon and intron abided by the role of GT…AG strictly. Follistatin gene encoded a 323-amino-acid preprotein. The preprotein contained signal domain, N-terminal domain, Follistatin domainⅠ, Follistatin domainⅡ, Follistatin domainⅢ, and C-terminal domain. Comparison of deduced amino acid sequences of flounder Follistatin with other vertebrate showed that Follistatin was highly conserved, the identity of domain was higher.Forecasting of Follistatin protein structure of flounder revealed that the model of flounder Follistatin was the same as that of zebrafish. The function of highly conserved cysteines might be maintaining the structure of Follistatin protein.2. Analysis of expression of Follistatin during flounder embryo development revealed that Follistatin began express in mesoderm cells at 26 hours after fertilization and continues express at 28, 30, 32, 34, 36, 38, 40and 42 hours after fertilization in ventral of somite , head, dorsal of somite cells. Double in situ hybridization revealed that the region where Follistatin and MyoD expressed partly overlapped at somite and cryosection of double in situ hybridization samples showed that the expression of the two genes intercrossed and partly overlapped. But Follistatin did not express in the...