Short-term, but not long-term feed restriction causes differential expression of leptins in Atlantic salmon.

Atlantic salmon was used to investigate the effect of long- and short-term dietary ration on the tissue expression levels of leptins. Compared to ad libitum fed fish (0.8-3kg), 6months of dietary restriction (60%) resulted in significantly lower body mass and adiposity, but did not produce a clear effect on the expression levels of either lepa1 or lepa2. For visceral adipose tissue, however, the long-term data indicated that season appeared to influence the levels of lepa1 expression of ad libitum fed fish, but not feed-restricted fish. By comparing the total levels of leptin mRNA expression to the tissue lipid contents, we found that only white muscle lepa1 showed the positive relation reported in mammals. The existence of a postprandial leptin response in Atlantic salmon parr was determined in fed and unfed parr over a 24h period. In contrast to other animals, lepa1 peaked in the unfed fish, initially in the white muscle at 6h, and subsequently in belly flap, liver and visceral adipose tissue at 9h. Only lepa2 in the visceral adipose tissue of fed fish showed a similar 9h peak, but at an order of magnitude lower than lepa1 in the unfed fish. These data reveal that short-term feed restriction causes a latent (6-9h) upregulation of lepa-type genes in the fatty tissues of Atlantic salmon, a finding that contrasts the mammalian response.

Leptin reduces Atlantic salmon growth through the central pro-opiomelanocortin pathway.

Leptin (Lep) is a key factor for the energy homeostasis in mammals, but the available data of its role in teleosts are not conclusive. There are large sequence differences among mammalian and teleost Lep, both at the gene and protein level. Therefore, in order to characterize Lep function in fish, the use of species-specific Lep is crucial. In this study, the cDNA sequence of salmon leptin a1 (lepa1) was used to establish a production protocol for recombinant salmon LepA1 (rsLepA1) in Escherichia coli, that enabled a final yield of 1.7 mg pure protein L⁻¹ culture. The effects of 20-day administration of rsLepA1 on growth and brain neuroendocrine peptide gene expression [npy, cart, agrp (-1 and -2), pomc (-a1, -a2, -a2s, and -b)] were studied in juvenile, immature Atlantic salmon (96.5±2.1g) fed a commercial diet to satiation. Intraperitoneal osmotic pumps were used to deliver rsLepA1 at four different concentrations (calculated pumping rates were 0, 0.1, 1.0 and 10 ng g⁻¹ h⁻¹). In the highest dosage group (10 ng g⁻¹ h⁻¹), the growth rate was significantly reduced, and pomc-a1 gene expression was higher than in controls. The results support the lipostatic hypothesis and suggest that sLepA1 reduces growth in Atlantic salmon by affecting food intake through the central pro-opiomelanocortin pathway.

Ontogeny of energy homeostatic pathways via neuroendocrine signaling in Atlantic salmon.

Leptin and ghrelin are known to regulate energy homeostasis via hypothalamic neuropeptide signaling in mammals. Recent studies have discovered that these hormones exist in teleosts, however, very little is known concerning their role during teleost ontogeny. Here, we have examined the steady state levels of leptins, ghrelins, their target neuropetides and several growth factors during Atlantic salmon development. Initial experiments revealed differential expression of leptin genes and ghrelin isoforms during embryogenesis. In larvae, equal upregulation of ghrl1 and ghrl2 was observed just prior to exogenous feeding while a surge of lepa1 occurred one week after first-feeding. Subsequent dissection of the embryos and larvae showed that lepa1, cart, pomca1, and agrp are supplied as maternal transcripts. The earliest zygotic expression was observed for lepa1 and cart at 320 day degrees. By 400 day degrees, this expression was localized to the head and coincided with upregulation of ghrl2 and npy. Over the hatching period growth factor signaling predominated. The ghrelin surge prior to first-feeding was exclusively localized in the internal organs and coincided with upregulation of npy and agrp in the head and agrp in the trunk. One week after exogenous feeding was established major peaks were detected in the head for lepa1 and pomca1 with increasing levels of cart, while lepa1 was also significantly expressed in the trunk. By integrating theses data into an ontogenetic model, we suggest that the mediation of Atlantic salmon energy homeostatic pathways via endocrine and neuropeptide signaling retains putative features of the mammalian system.