Production of recombinant leptin and its effects on food intake in rainbow trout (Oncorhynchus mykiss).
Leptin is a key factor for the regulation of food intake and energy homeostasis in mammals, but information regarding its role in teleosts is still limited. There are large differences between mammalian and teleost leptin at both gene and protein levels, and in order to characterize the function of leptin in fish, preparation of species-specific leptin is therefore a key step. In this study, full-length cDNA coding for rainbow trout leptin was identified. In spite of low amino acid sequence similarity with other animals, leptin is highly conserved between trout and salmon (98.7%). Based on the cDNA, we produced pure recombinant trout leptin (rt-leptin) in E. coli, with a final yield of 20 mg/L culture medium. We then examined the effects of intraperitoneal (IP) injection of rt-leptin on feeding behavior and gene expression of hypothalamic NPY and POMCs (POMC A1, A2 and B) in a short-term (8 h) experiment. The rt-leptin suppressed food intake and led to transient reduction of NPY mRNA levels, while the expression of POMCs A1 and A2, was elevated compared with vehicle-injected controls. These results for rainbow trout are the first that describe a physiological role of leptin using a species-specific orthologue in teleosts, and they suggest that leptin suppresses food intake mediated by hypothalamic regulation. This anorexic effect is similar to that observed in mammals and frogs and supports that the neuroendocrine pathways that control feeding by leptin are ancient and have been conserved through evolution.
Murashita
Koji
K
Nansei Station, National Research Institute of Aquaculture, Fisheries Research Agency, 422-1 Nakatsuhamaura, Minami-ise, Mie 516-0193, Japan.
Uji
Susumu
S
Yamamoto
Takeshi
T
Rønnestad
Ivar
I
Kurokawa
Tadahide
T
D000595:Amino Acid Sequence; D000818:Animals; D004435:Eating / Q000187:drug effects*; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D007276:Injections, Intraventricular; D020738:Leptin / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000235:genetics; D017686:Oncorhynchus mykiss / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011333:Pro-Opiomelanocortin / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D029941:Fish Proteins; D020738:Leptin; D009478:Neuropeptide Y; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011333:Pro-Opiomelanocortin