FGF21 mimics a fasting-induced metabolic state and increases appetite in zebrafish

Fibroblast growth factor 21 (FGF21) is a member of the FGF superfamily that acts in an endocrine manner. FGF21 is a key regulator of energy balance and metabolism in mammals, and has emerged as a therapeutic potential for treating obesity and diabetes. Here, we report that mRNAs encoding FGF21 and its receptors are widely distributed within the zebrafish tissues and are importantly modulated by fasting (decreased in brain and liver, and increased in gut). FGF21 stimulates food intake in zebrafish, likely in part by modulating brain npy/agrp and nucb2/nesfatin-1 and gut ghrelin and cck mRNA expression. In accordance with this orexigenic role, the expression of FGF21 and its receptors were observed to increase preprandially and decrease post-feeding in the foregut and/or liver. Finally, we found important evidence in favor of a role for FGF21 in regulating glucose and lipid metabolism in the zebrafish liver in a way that mimics a fasting metabolic stateNatural Sciences and Engineering Research Council (NSERC) of Canada, and the University of Saskatchewan Centennial Enhancement Chair in Comparative Endocrinology to S. Unniappan | Ref. 413566–2017-RGPINXunta de Galicia | Ref. ED481B 2017/11

Feeding and food availability modulate brain-derived neurotrophic factor, an orexigen with metabolic roles in zebrafish

Emerging fndings point to a role for brain-derived neurotrophic factor (BDNF) on feeding in mammals. However, its role on energy balance is unclear. Moreover, whether BDNF regulates energy homeostasis in non-mammals remain unknown. This research aimed to determine whether BDNF is a metabolic peptide in zebrafsh. Our results demonstrate that BDNF mRNAs and protein, as well as mRNAs encoding its receptors trkb2, p75ntra and p75ntrb, are detectable in the zebrafsh brain, foregut and liver. Intraperitoneal injection of BDNF increased food intake at 1, 2 and 6 h post-administration, and caused an upregulation of brain npy, agrp and orexin, foregut ghrelin, and hepatic leptin mRNAs, and a reduction in brain nucb2. Fasting for 7 days increased bdnf and p75ntrb mRNAs in the foregut, while decreased bdnf, trkb2, p75ntra and p75ntrb mRNAs in the brain and liver. Additionally, the expression of bdnf and its receptors increased preprandially, and decreased after a meal in the foregut and liver. Finally, we observed BDNF-induced changes in the expression and/ or activity of enzymes involved in glucose and lipid metabolism in the liver. Overall, present results indicate that BDNF is a novel regulator of appetite and metabolism in fsh, which is modulated by energy intake and food availabilityNatural Sciences and Engineering Research Council (NSERC) of Canada | Ref. RGPIN-2017-04956Xunta de Galicia | Ref. ED481B 2017/11

Evidence of gastrointestinal sensing and gut-brain communication in rainbow trout (Oncorhynchus mykiss) in response to the aqueous extract of fishmeal and its free amino acid fraction

Using rainbow trout (Oncorhynchus mykiss) as a model, we aimed to obtain information about the gastrointestinal tract (GIT) amino acid sensing capacity and hormone production along regions of the GIT, in response to proline (Pro), to a solution of free amino acids (FAA) mimicking the composition of a fishmeal (FM) aqueous extract (FM-FAA), or to the whole FM aqueous extract (FM-AQE). In addition, we evaluated central responses (in hypothalamus) in mechanisms regulating food intake, 2 h following intragastric administration of these treatments. The presence of Pro in the GIT elicited changes in amino acid sensing systems and in the production of GIT hormones, especially in the more proximal regions in parallel with an anorectic response in hypothalamus. The intragastric administration of FM-AQE induced increased production of the anorectic hormones peptide tyrosine-tyrosine (PYY) and cholecystokinin (CCK) that occurred 20 min post-treatment in the proximal and middle intestine of this treatment. These changes occurred in parallel with an anorectic response in the hypothalamus 2 h post-treatment. The treatment with FM-FAA elicited a comparable anorectic response in the hypothalamus at 2 h post-treatment, which was associated however with a more complex response in the GIT. This included a comparable increased production of the anorectic hormones PYY and CCK in the proximal and middle intestine, but also a decreased production of the orexigenic hormone ghrelin (GHRL) in the stomach, 20 min after FM-FAA administration. These effects were also accompanied by some changes in parameters related to amino acid sensing systems mediated by receptors, which were not observed in the FM-AQE treatment. Overall, results indicate that all treatments elicited a response in elements of gut sensing mechanisms and gut-brain axis, despite important differences in the specific genes (likely having different substrate specificities), GIT areas and times in which responses were observedXunta de Galicia | Ref. ED431B 2022/01Ministerio de Universidades | Ref. FPU19/00122Agencia Estatal de Investigación | Ref. IJC2019-039166-IFinanciado para publicación en acceso aberto: Universidade de Vigo/CISU

Fatty acids of different nature differentially modulate feed intake in rainbow trout

Feed intake is subjected to a complex regulation involving a plethora of signals, among which nutrients stand as one of the most important. In mammals, the gastrointestinal tract is able to sense nutrients in the lumen, and respond with the release of signaling molecules that ultimately modulate brain circuits governing appetite, resulting in decreased/increased feeding. Whether equivalent mechanisms operate in fish remains unknown. In a recent study, we described that the gastrointestinal tract of rainbow trout contains several sensors for free fatty acids (FAs), and that the luminal presence of FAs of different length and degree of unsaturation modulates the levels of key gastrointestinal hormones involved in feed intake regulation. In this study, our aim was to characterize the impact of such a luminal presence of FAs on brain appetite-regulatory centers, as well as its effects on rainbow trout feed intake. Major results from this study demonstrated that: (i) FAs of different length and degree of unsaturation [medium-chain (MCFAs, octanoate), long-chain (LCFAs, oleate), long-chain polyunsaturated (PUFA, α-linolenate), and short-chain (SCFA, butyrate) FAs] differentially modulate feed intake levels when administered intragastrically, (ii) intragastrically-administered FAs modulate the phosphorylation status of appetite-related transcription factors, as well as mRNA levels of key appetite-regulating neuropeptides, in the hypothalamus and/or telencephalon, (iii) luminal presence of FAs results in changes in the central abundance of mRNAs encoding gastrointestinal hormone receptors, and (vi) luminal FA-derived central changes in neuropeptide mRNAs are not observed (or are lessened) in vagotomized fish. Together, these results provide comprehensive evidence in favor of a gut-brain axis in fish. In addition, we observed different responses in terms of feed intake regulation depending on the type of fatty acid administered into the lumen, which is very relevant for aquaculture considering differences in fatty acid composition in aquafeedsAgencia Estatal de Investigación | Ref. PID2019-103969RB-C31Agencia Estatal de Investigación | Ref. IJC2019-039166-IMinisterio de Universidades | Ref. FPU19/00122Ministerio de Educación, Cultura y Deporte | Ref. FPU16/0004

Fatty acid sensing in the gastrointestinal tract of rainbow trout: different to mammalian model?

It is well established in mammals that the gastrointestinal tract (GIT) senses the luminal presence of nutrients and responds to such information by releasing signaling molecules that ultimately regulate feeding. However, gut nutrient sensing mechanisms are poorly known in fish. This research characterized fatty acid (FA) sensing mechanisms in the GIT of a fish species with great interest in aquaculture: the rainbow trout (Oncorhynchus mykiss). Main results showed that: (i) the trout GIT has mRNAs encoding numerous key FA transporters characterized in mammals (FA transporter CD36 -FAT/CD36-, FA transport protein 4 -FATP4-, and monocarboxylate transporter isoform-1 -MCT-1-) and receptors (several free FA receptor -Ffar- isoforms, and G protein-coupled receptors 84 and 119 -Gpr84 and Gpr119-), and (ii) intragastrically-administered FAs differing in their length and degree of unsaturation (i.e., medium-chain (octanoate), long-chain (oleate), long-chain polyunsaturated (α-linolenate), and short-chain (butyrate) FAs) exert a differential modulation of the gastrointestinal abundance of mRNAs encoding the identified transporters and receptors and intracellular signaling elements, as well as gastrointestinal appetite-regulatory hormone mRNAs and proteins. Together, results from this study offer the first set of evidence supporting the existence of FA sensing mechanisms n the fish GIT. Additionally, we detected several differences in FA sensing mechanisms of rainbow trout vs. mammals, which may suggest evolutionary divergence between fish and mammals.Xunta de Galicia | Ref. ED431B 2022/01Agencia Estatal de Investigación española | Ref. PID2019-103969RB-C31Ministerio de Educación, Cultura y Deporte | Ref. FPU19/00122Ministerio de Educación, Cultura y Deporte | Ref. FPU16/00045Ministerio de Ciencia e Innovación | Ref. IJC2019-039166-

Impact of feeding diets with enhanced vegetable protein content and presence of umami taste-stimulating additive on gastrointestinal amino acid sensing and feed intake regulation in rainbow trout

The regulation of feed intake in fish is dependent upon different neuroendocrine and metabolic mechanisms including amino acid sensing in the gastrointestinal tract (GIT). However, there is little information regarding the impact of diets on such mechanisms. Therefore, in this study, we fed rainbow trout (Oncorhynchus mykiss) with 3 diets: a diet with high content of fishmeal and low content of soybean protein concentrate (SPC) (HF), a diet with a reduced content of fishmeal and high content of SPC (LF), and the LF diet supplemented with an umami tastestimulating additive (LFU). Fish were fed ad libitum once a day for 4 weeks, with no significant differences being registered in feed intake among groups. At the end of the feeding trial, we collected samples of different areas of the GIT (stomach, proximal and distal intestine) and hypothalamus at different times: after 48 h of fasting (time 0), and 1 h, 4 h, and 24 h after feeding. We evaluated the activity of pepsin in the stomach and trypsin and chymotrypsin in the proximal intestine, as well as mRNA abundance of transcripts encoding amino acid transporters and taste receptors, intracellular signalling molecules, and hormones. Moreover, we assessed the hypothalamic mRNA abundance of neuropeptides involved in feed intake regulation. Feeding rainbow trout with LF did not result in marked alterations in parameters related to digestive function and amino acid sensing in the rainbow trout GIT, nor in the expression of gastrointestinal hormones (except cck) and hypothalamic neuropeptides. In contrast, supplementation of the LF diet with an umami taste-stimulating additive resulted in a general improvement of digestive or absorptive function (increased protein, dry matter and energy digestibility, and earlier peak in plasma amino acid levels) and activation of gut-brain axis mechanisms involved in feed intake regulation through the transcriptional activation of amino acid transporters, taste receptors, signalling molecules, and hormones. These results demonstrate that the dietary inclusion of umami receptor stimulants has the potential to improve fish physiological responses to the rise in levels of vegetable protein in the diet.Xunta de Galicia | Ref. GPC-ED431B 2022/01Ministerio de Universidades | Ref. FPU19/00122Agencia Estatal de Investigación | Ref. IJC2019-039166-IUniversidade de Vigo/CISU

Leptin signalling in teleost fish with emphasis in food intake regulation

Leptin, the product of the obese (ob or Lep) gene, was first cloned in teleost fish in 2005, more than a decade after its identification in mammals. This was because bony fish and mammalian leptins share a very low amino acid sequence identity, which suggests different functionality of the leptin system in fish compared to that of mammals. Indeed, major differences are evident between the mammalian and fish leptin system. Thus, for instance, mammalian leptin is synthesized and released by the adipose tissue in response to the amount of fat depots, while several tissues (mainly the liver) are the main sources of leptin in fish, whose determining factors of production are still unclear. In mammals, the main physiological role for leptin is its involvement in the maintenance of energy balance by decreasing food intake and increasing energy expenditure, although a wide variety of actions have been attributed to this hormone (e.g., regulation of lipid and carbohydrate metabolism, reproduction and immune functions). In fish, available literature also points towards a multifunctional nature for leptin, although knowledge on its functions is limited. In this review, we offer an overview of teleostean leptin structure and mechanism of action, and discuss the available knowledge on the role of this hormone in food intake regulation in teleost fish, aiming to provide a comparative overview between the functioning of the teleostean and mammalian leptin systemsFinanciado para publicación en acceso aberto: Universidade de Vigo/CISUGAgencia Estatal de Investigación | Ref. PID2019-103969RB-C31Xunta de Galicia | Ref. ED431B 2019/37Xunta de Galicia | Ref. ED481B 2017/11

First evidence for the presence of amino acid sensing mechanisms in the fish gastrointestinal tract

This study aimed to characterize amino acid sensing systems in the gastrointestinal tract (GIT) of the carnivorous fish model species rainbow trout. We observed that the trout GIT expresses mRNAs encoding some amino acid receptors described in mammals [calcium-sensing receptor (CaSR), G protein-coupled receptor family C group 6 member A (GPRC6A), and taste receptors type 1 members 1 and 2 (T1r1, T1r2)], while others [taste receptor type 1 member 3 (T1r3) and metabotropic glutamate receptors 1 and 4 (mGlur1, mGlur4)] could not be found. Then, we characterized the response of such receptors, as well as that of intracellular signaling mechanisms, to the intragastric administration of L-leucine, L-valine, L-proline or L-glutamate. Results demonstrated that casr, gprc6a, tas1r1 and tas1r2 mRNAs are modulated by amino acids in the stomach and proximal intestine, with important differences with respect to mammals. Likewise, gut amino acid receptors triggered signaling pathways likely mediated, at least partly, by phospholipase C β3 and β4. Finally, the luminal presence of amino acids led to important changes in ghrelin, cholecystokinin, peptide YY and proglucagon mRNAs and/or protein levels. Present results offer the first set of evidence in favor of the existence of amino acid sensing mechanisms within the fish GI