title: [Molecular cloning, tissue distribution and the expression in the regulation of food intake of prepro-orexin in Nile tilapia (Oreochromis niloticus)].
abstract: We cloned the full length of tilapia prepro-orexin cDNA using RT-PCR and rapid-amplification of cDNA ends (RACE). The full-length of prepro-orexin cDNA was 648 bp containing an open reading frame of 423 bp. The 140 amino acid prepro-orexin protein included a 37 AA signal peptide, a 43 AA Orexin-A and, and 28 AA Orexin-B and the end of the 32 AA peptide of unknown function. The expression of prepro-orexin on tissue distribution, peri-prandial changes, starvation and re-feeding were quantified by real-time PCR. We found that prepro-orexin mRNA was present in all tissues tested and that the highest level was observed in hypothalamus. Expression levels were significantly higher at mealtime (0 h) than before (-2 h, -1 h) and after (+1 h, +2 h) mealtime. Fasting for 2, 4, 6 and 8 d caused significant increases in prepro-orexin mRNA expression in the hypothalamus, and after re-feeding, expression levels of prepro-o rexin mRNA returned to the same level compared to that in the fed group.
authors.affiliation: State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, China.
authors.forename: Wen-Bo
authors.initials: WB
authors.lastname: Chen
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Ya-Lu
authors.initials: YL
authors.lastname: Zhou
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Dong
authors.forename: Hao-Ran
authors.initials: HR
authors.lastname: Lin
authors.forename: Wen-Sheng
authors.initials: WS
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D023681:Cichlids / Q000145:classification / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D004435:Eating; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000068797:Orexins; D010802:Phylogeny; D016415:Sequence Alignment; D014018:Tissue Distribution; D014714:Vertebrates / Q000145:classification / Q000235:genetics
chemical_list: D029941:Fish Proteins; C000596229:HCRT protein, human; D047908:Intracellular Signaling Peptides and Proteins; D009479:Neuropeptides; D000068797:Orexins