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The melanocortin system is a key regulator of appetite and food intake in vertebrates. This system includes the neuropeptides neuropeptide y (NPY), agouti-related peptide (AGRP), cocaine- and amphetamine-regulated transcript (CART), and pro-opiomelanocortin (POMC). An important center for appetite control in mammals is the hypothalamic arcuate nucleus, with neurons that coexpress either the orexigenic NPY/AGRP or the anorexigenic CART/POMC neuropeptides. In ray-finned fishes, such a center is less characterized. The Atlantic salmon (Salmo salar) has multiple genes of these neuropeptides due to whole-genome duplication events. To better understand the potential involvement of the melanocortin system in appetite and food intake control, we have mapped the mRNA expression of npy, agrp, cart, and pomc in the brain of Atlantic salmon parr using in situ hybridization. After identifying hypothalamic mRNA expression, we investigated the possible intracellular coexpression of npy/agrp and cart/pomc in the tuberal hypothalamus by fluorescent in situ hybridization. The results showed that the neuropeptides were widely distributed, especially in sensory and neuroendocrine brain regions. In the hypothalamic lateral tuberal nucleus, the putative homolog to the mammalian arcuate nucleus, npya, agrp1, cart2b, and pomca were predominantly localized in distinct neurons; however, some neurons coexpressed cart2b/pomca. This is the first demonstration of coexpression of cart2b/pomca in the tuberal hypothalamus of a teleost. Collectively, our data suggest that the lateral tuberal nucleus is the center for appetite control in salmon, similar to that of mammals. Extrahypothalamic brain regions might also be involved in regulating food intake, including the olfactory bulb, telencephalon, midbrain, and hindbrain., (© 2022 The Authors. The Journal of Comparative Neurology published by Wiley Periodicals LLC.)

Episodes of elevated temperature, combined with lower feed availability, are among the predicted scenarios of climate change representing a challenge for coral reef fish. We investigated the response of clownfish (Amphiprion ocellaris) to a scenario in which it received a single meal to satiety after 48 h fasting at 32 °C (climate change scenario) and 28 °C (control). We analysed the metabolic rate (MR), feed intake, gut transit, and expression of selected brain neuropeptides and one receptor believed to be involved in appetite control. Fish at 32 °C ingested 17.9% less feed and had a faster gut transit than did fish at 28 °C. MR in the unfed fish was 31% higher at 32 °C compared to 28 °C. In the fed fish, postprandial MR at 28 °C was 30% higher compared to that of unfed fish, while at 32 °C it was only 15% higher. The expression of agrp1 did not differ between unfed and refed fish. The levels of both pomca and mc4r increased immediately after the meal and subsequently declined, suggesting a possible anorexic role for these genes. Notably, this pattern was accelerated in fish kept at 32 °C compared with that in fish kept at 28 °C. The dynamics of these changes in expression correspond to a faster gut transition of ingested feed at elevated temperatures. For both agrp2 and pomcb there was an increase in expression following feeding in fish maintained at 32 °C, which was not observed in fish kept at 28 °C. These results suggest that low feed availability and elevated temperature stimulate anorexigenic pathways in clownfish, resulting in significantly lower feed intake despite the temperature-induced increase in metabolic rate. This may be a mechanism to ameliorate the decrease in aerobic scope that results from higher temperatures., Competing Interests: Declaration of Competing Interest Ref.: Ms. No. CBPA-D-22-00190. Metabolic rates, feed intake, appetite control, and gut transit of clownfish Amphiprion ocellaris exposed to increased temperature and limited feed availability. Comparative Biochemistry and Physiology, Part A. All authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The clown anemonefish (Amphiprion ocellaris) is a common model species in studies assessing the impact of climate changes on tropical coral fish physiology, metabolism, growth, and stress. However, the basic endocrine principles for the control of food intake and energy homeostasis, under normal and elevated sea temperatures, in this species remain unknown. In this work, we studied food intake and growth in clown anemonefish reared at different temperatures and with different food availability. We also analyzed expression of genes in the melanocortin system, which is believed to be involved in the control of appetite and feeding behavior. These were two paralogues of pomc: pomca and pomcb; two paralogs of agrp: agrp1 and agrp2; and one mc4r-like. Groups of juvenile clown anemonefish were exposed to four experimental treatments combining (orthogonal design) two rearing temperatures: 28 °C (T28; normal) and 32 °C (T32; high) and two feeding regimes: one (1 M; 08:00) or three (3 M; 08:00, 12:00, 15:00) meals per day, fed to satiety by hand. The results showed that high temperature (T32) did not affect the average growth rate but induced a stronger asymmetrical individual body weight of the fish within the population (tank). Lower feeding frequency (1 M) resulted in lower growth rates at both rearing temperatures. Fish reared at high temperature had higher total daily food intake, which correlated with a lower expression of pomca, supporting an anorexigenic role of this gene. High temperature combined with restricted feeding induced higher agrp1 levels and resulted in a higher food intake in the morning meal compared to the control. This supports an orexigenic role for agrp1. mRNA levels of agrp2 responded differently from agrp1, supporting different roles for the paralogues. Levels of mc4r-like inversely correlated with fish body weight, indicating a possible size/stage dependence of gene expression. In conclusion, our results indicate that the melanocortin system is involved in adjusting appetite and food intake of clown anemonefish in response to elevated temperature and low food availability., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Similar to many marine teleost species, Atlantic cod undergo remarkable physiological changes during the early life stages with concurrent and profound changes in feeding biology and ecology. In contrast to the digestive system, very little is known about the ontogeny and the localization of the centers that control appetite and feed ingestion in the developing brain of fish. We examined the expression patterns of three appetite regulating factors (orexigenic: neuropeptide Y, NPY; prepro-orexin, pOX and anorexigenic: cocaine- and amphetamine-regulated transcript, CART) in discrete brain regions of developing Atlantic cod using chromogenic and double fluorescent in situ hybridization. Differential temporal and spatial expression patterns for each appetite regulator were found from first feeding (4 days post hatch; dph) to juvenile stage (76 dph). Neurons expressing NPY mRNA were detected in the telencephalon (highest expression), diencephalon, and optic tectum from 4 dph onward. CART mRNA expression had a wider distribution along the anterior-posterior brain axis, including both telencephalon and diencephalon from 4 dph. From 46 dph, CART transcripts were also detected in the olfactory bulb, region of the nucleus of medial longitudinal fascicle, optic tectum and midbrain tegmentum. At 4 and 20 dph, pOX mRNA expression was exclusively found in the preoptic region, but extended to the hypothalamus at 46 and 76 dph. Co-expression of both CART and pOX genes were also observed in several hypothalamic neurons throughout larval development. Our results show that both orexigenic and anorexigenic factors are present in the telencephalon, diencephalon and mesencephalon in cod larvae. The telencephalon mostly contains key factors of hunger control (NPY), while the diencephalon, and particularly the hypothalamus may have a more complex role in modulating the multifunctional control of appetite in this species. As the larvae develop, the overall progression in temporal and spatial complexity of NPY, CART and pOX mRNAs expression might be correlated to the maturation of appetite control regulation. These observations suggest that teleost larvae continue to develop the regulatory networks underlying appetite control after onset of exogenous feeding.

The first studies that identified leptin and its receptor (LepR) in mammals were based on mutant animals that displayed dramatic changes in body-weight and regulation of energy homeostasis. Subsequent studies have shown that a deficiency of leptin or LepR in homoeothermic mammals results in hyperphagia, obesity, infertility and a number of other abnormalities. The physiological roles of leptin-mediated signaling in ectothermic teleosts are still being explored. Here, we produced medaka with homozygous LepR gene mutation using the targeting induced local lesions in a genome method. This knockout mutant had a point mutation of cysteine for stop codon at the 357th amino acid just before the leptin-binding domain. The evidence for loss of function of leptin-mediated signaling in the mutant is based on a lack of response to feeding in the expression of key appetite-related neuropeptides in the diencephalon. The mutant lepr−/− medaka expressed constant up-regulated levels of mRNA for the orexigenic neuropeptide Ya and agouti-related protein and a suppressed level of anorexigenic proopiomelanocortin 1 in the diencephalon independent of feeding, which suggests that the mutant did not possess functional LepR. Phenotypes of the LepR-mutant medaka were analyzed in order to understand the effects on food intake, growth, and fat accumulation in the tissues. The food intake of the mutant medaka was higher in post-juveniles and adult stages than that of wild-type (WT) fish. The hyperphagia led to a high growth rate at the post-juvenile stage, but did not to significant alterations in final adult body size. There was no additional deposition of fat in the liver and muscle in the post-juvenile and adult mutants, or in the blood plasma in the adult mutant. However, adult LepR mutants possessed large deposits of visceral fat, unlike in the WT fish, in which there were none. Our analysis confirms that LepR in medaka exert a powerful influence on the control on food intake. Further analyses using the mutant will contribute to a better understanding of the role of leptin in fish. This is the first study to produce fish with leptin receptor deficiency.

Following feeding of a single meal to Atlantic salmon, the temporal changes in the brain mRNA expression of neuropeptide y (npy), cocaine-amphetamine regulated transcript (cart), peptide yy (pyy), two isoforms of agouti-related protein (agrp), two isoforms of cholecystokinin (cck), and four isoforms of proopiomelanocortin (pomc) were assessed by q-PCR. In the course of 24h post-feeding (hpf), several of the brain neuropeptides displayed changes in mRNA expression compared to an unfed control group, indicating that food intake and processing affect the regulation of expression of these genes in Atlantic salmon. Expression of cart, cck-l, pomc-a1 and pomc-b all increased within 3h of feeding, while most of the feed was still in the stomach, suggesting that these neuropeptides play central anorexigenic roles similar to those described in higher vertebrates, including determining meal intervals. On the other hand, the npy and agrp isoforms which have been described as playing orexigenic roles in mammals, showed an opposite response in salmon and both were elevated in the first 3h after feeding. The different isoforms of cck, agrp and pomc had different mRNA expression patterns, which indicate specific roles related to feeding regulation. The minimal effect of feeding and digestion on pyy expression in the brain indicates that PYY plays a minor role in the central control of short-term food intake in Atlantic salmon., (Copyright © 2011 Elsevier Inc. All rights reserved.)

One of the main gastrointestinal hormones, cholecystokinin (CCK), was studied in order to advance understanding of the control of the digestive process in Atlantic cod Gadus morhua larvae after onset of first feeding. Larvae were fed either natural zooplankton or enriched rotifers in similar rearing systems and sampled from hatching to 22 days post-hatch (dph). CCK was visualized by immunohistochemistry and the first CCK-immunoreactive (IR) cells were detected at 8 dph corresponding to 6 days after first feeding. The CCK-IR cells were mostly found in the anterior midgut, and the number of CCK-IR cells was lower in the posterior midgut. They were also present in the hindgut of some of the larvae, but not in the foregut. No clear differences were found in the ontogenetic appearance and the distribution pattern of CCK-IR cells between the two dietary treatments. This indicates that the onset of CCK production in the gut as well as the spatial distribution of the CCK-IR cells is not differentially affected by these diets. To what extent the hormone production itself is influenced by dietary factors needs to be studied by more sensitive methods. [ABSTRACT FROM AUTHOR]

Abstract: For the majority of marine fish larvae a fully developed digestive tract, including gastric digestion, is acquired weeks to months (depending on species) after onset of exogenous feeding. Still, the processing capacity (capability to degrade and absorb dietary nutrients) of the larval gut is sufficient to support fast larval growth by digesting prey naturally available in the sea. However, the physiological constraints of the gut with respect to digestion of cultivated live prey and particularly formulated starter feeds still remain to be elucidated. This paper reviews some recent findings in the areas of control and efficiency of digestive function of marine fish larvae. For studies on the hormonal control, the peptide hormone cholecystokinin (CCK) has been targeted, since it is believed to play an important role in controlling digestion in vertebrates. Recent work on the processing capacity include studies of the digestibility and transfer kinetics of macronutrients from live prey and experimental work on absorption of protein, peptides and free amino acids from the lumen of the digestive tract into the tissues of larval fish and how this changes during ontogeny. The molecular and in vitro characterization of transporters is currently being integrated with ongoing in vivo studies. [Copyright &y& Elsevier]

Leucine has been identified to modulate feed intake and energy homeostasis in fish as in other vertebrates. Under allostatic conditions, energy expenditure may change, and adjustments to the processes that govern the energy homeostatic system may be necessary. We investigated the responsiveness of appetite-related neuropeptides involved in feed intake regulation in Atlantic salmon (Salmo salar) reared with high (35 g/kg leucine) or control (27.3 g/kg leucine) leucine-supplemented diets and/or under chronic stressor conditions (chasing) for eight weeks. We also analysed the response of amino acid transporters potentially involved in uptake of branched-chain amino acids (BCAA), including leucine, into areas of the brain where nutrient sensors may signal locally or to other areas involved in appetite control. At the end of the experiment, all fish were subjected to a novel-acute stressor (confinement). Our results show that fish fed with high leucine diet had a lower feed intake, growth, and hepatosomatic index (HSI) when compared to fish fed control leucine diet. In addition, increased mRNA expression of amino acid solute carrier (slc) genes in the diencephalon, and genes related to appetite control, such as proopiomelanocortin a1 (pomca1), in both the diencephalon and telencephalon, imply their involvement in leucine anorectic effect. Stress, as high leucine, reduced feed intake, growth and HSI of fish fed control or high leucine diet and antagonized the high leucine effect on the slc genes mRNA expression. An increase of neuropeptide y a1 (npya1) was observed both due to high dietary leucine and/or stress treatment which may represent a compensatory regulatory mechanism with the aim to reverse the decrease in feed intake. In summary, our results confirm an anorectic role of high dietary leucine via the activation of amino acid sensing mechanisms in the brain. Further, corticotropin-releasing hormone 1 b1 (crh1b1) and npya1 showed to play a role in the regulation of appetite in Atlantic salmon under stress conditions and/or high leucine levels. • High dietary leucine acts as anorectic factor in the control of energy balance in Atlantic salmon. • Both slc transporters and pomca1 neuropeptide are involved in the leucine anorexigenic control. • The same mechanisms do not seem to be responsible for such disruption under stressed conditions. • Instead, crf1b1 and npya1 play a role in the regulation of appetite under stressful conditions. [ABSTRACT FROM AUTHOR]