RESUMO
The full-length neuropeptide Y (NPY) cDNA of Chinese perch Siniperca chuatsi was 704 bp in length, and contained a 300 bp open reading frame encoding a prepro-NPY with 99 amino acids. The predicted prepro-NPY peptide contained a putative signal peptide of 28 amino acids and a mature NPY of 36 amino acids, followed by the proteolytic processing site Gly-Lys-Arg and 35 amino acids that comprise the C-terminal peptide of NPY. Amino acid alignment and phylogenetic analysis indicate that the predicted Chinese perch prepro-NPY (composed of 99 amino acids) had high identities to the prepro-NPY of acanthomorph fishes (93-95%), whereas it had much lower identities to the prepro-NPY (composed of 96 or 97 amino acids) of cyprinid fishes (59-60%) or mammals (57-63%). Chinese perch NPY gene consists of four exons and three introns. The ratio of intron 2 to intron 3 was over 14 in Chinese perch NPY gene, whereas this ratio was below 4 in zebrafish and mammalian NPY gene. The total size of the Chinese perch NPY gene was 2223 bp, which was only about 28% of the size of NPY gene in higher vertebrate. Analysis of a 1622 bp promoter region of Chinese perch NPY gene, revealed a typical TATA box, a GC box and an untypical CAAT box, located at 84 bp, 101 bp and 303 bp upstream of the start codon respectively. Three STAT binding site-like elements (TCCAGTA) which were necessary for the leptin-induced transcriptional control of rat NPY gene were identified. In consistence to the effect of cortisol on fish brain NPY mRNA expression, four glucocorticoid-responsive elements were detected. Besides the highest expression in brain, substantial level of Chinese perch NPY mRNA expression was detected in the spleen and liver, and trace level of NPY mRNA expression was also detected in the adipose tissue, intestine and muscle. These results indicated that Chinese perch NPY might be involved in the food intake control by leptin and cortisol system, and diversification of NPY signaling should exist between acanthomorph fishes and cyprinid fishes as well as mammals.
