Nuria de Pedro

Neuropeptide Y has a stimulatory action on feeding behavior in goldfish (Carassius auratus).

The purpose of the present study was to elucidate the possible role of neuropeptide Y (NPY) in the feeding regulation in fish. We examined the effects of intracerebroventricular (i.c.v.) or intraperitoneal (i.p.) neuropeptide Y administration on food intake in satiated goldfish, at different time intervals postinjection (0-2, 2-8 and 0-8 h). Food intake was significantly increased by i.c.v. administered neuropeptide Y (1 microg) at 2 h postinjection, while no significant differences in food intake were observed after i.p. treatment. The neuropeptide Y receptor antagonist, neuropeptide Y-(27-36), totally counteracted the stimulatory action of neuropeptide Y on feeding. The possible involvement of neuropeptide Y in the eating behavior evoked by food deprivation has been investigated. Food deprivation by either 24 or 72 h significantly increased feeding, and the neuropeptide Y receptor antagonist attenuated such feeding stimulation. From our findings, we suggest, first, that neuropeptide Y is involved in feeding central regulation in goldfish, acting via specific neuropeptide Y receptors, and second, that hypothalamic neuropeptide Y would be released in response to food deprivation, contributing to generate the consequent eating behavior stimulation in Carassius auratus.

Inhibitory Effect of Serotonin on Feeding Behavior in Goldfish: Involvement of CRF

The possible action of 5-HT on feeding behavior in goldfish has been studied. Food intake was significantly reduced by intracerebroventricular (ICV) injection of serotonin (5-HT, 10 microg) at 2 h postinjection. After peripheral (intraperitoneal) administration of 5-HT (1 and 10 microg/g bw), no significant modifications in food intake were detected. Thus, it can be concluded that there is a central anoretic action of 5-HT in teleost fish. Taking in mind the inhibitory effect of corticotropin releasing factor (CRF) on feeding in teleosts and the interactions between 5-HT and CRF described in mammals, we investigated the possible involvement of CRF as mediator of the 5-HT anoretic action in goldfish. The ICV pretreatment with alpha-Helical CRF[9-41](20 microg) partially blocked the inhibitory effect of 5-HT on food consumption in goldfish. These results show that CRF mediates, at least in part, the 5-HT-induced feeding inhibition in goldfish. On the other hand, the alterations in hypothalamic indoleamines content evoked by ICV treatments would suggest that the activation of CRF neurons by 5-HT appears to inhibit hypothalamic serotoninergic transmission, supporting the intermediate role of this neuropeptide in the central anoretic effect of 5-HT in goldfish.

Role of corticotropin-releasing factor (CRF) as a food intake regulator in goldfish

The effect of intraperitoneal or intracerebroventricular corticotropin-releasing factor (CRF) administration on food intake has been studied in goldfish after 24 h of food deprivation. Food intake was evaluated at different time periods after injection, 0-2, 2-8, and 0-8 h. The 1 and 2 micrograms doses of CRF intracerebroventricularly administered induced a reduction in food intake during the first 2 h, although an increased feeding was observed in the next 6 h. The higher dose of CRF used in this study (3.3 micrograms) increased cumulative food intake at 8 h postinjection. However, intraperitoneal injection of 1 microgram CRF did not modify food intake in any of the studied intervals. These results suggest that CRF may play a role in central regulation of feeding behavior intake in goldfish, and show that CRF effects are time- and dose-dependent.

Leptins and leptin receptor expression in the goldfish (Carassius auratus). Regulation by food intake and fasting/overfeeding conditions

Leptin is a hormone involved in feeding and body weight regulation in vertebrates, but the relationship between energy status and leptin has not been clearly established in fish. The aim of this study was to investigate in a teleost, the goldfish (Carassius auratus), the tissue expression pattern of two leptins (gLep-aI and gLep-aII) and leptin receptor (gLepR); and the effect of feeding on expression of these genes. Leptin system expression in goldfish was firstly analyzed in fish under overfeeding (2 weeks) or fasting (1 week), and secondly, at different postfeeding times (0, 3, 6, 9 and 12h). Goldfish has two Lep-a paralog genes, gLep-aI was widely expressed in central and peripheral tissues, whereas gLep-aII was preferentially expressed in brain. This different distribution pattern of leptins suggests that they can play different physiological roles in goldfish. The gLepR mRNA was ubiquitous expressed, with the highest expression in the telencephalon and hypothalamus. No significant differences in the leptin system expression were found among control, overfed and fasting groups, suggesting an apparent lack of correlation between nutritional status and leptin system in goldfish. Hepatic expression of gLep-aI significantly increased 9h after feeding time, while hypothalamic leptin system expression did not change after feeding. In summary, leptin in goldfish could signal short-term changes in food intake, as postprandial satiety, but seems to be independent of fasting/overfeeding conditions in this teleost. The widespread distribution of leptins and leptin receptor in goldfish strongly supports that this hormone may have pleitropic actions in fish.

The galanin-induced feeding stimulation is mediated via α2-adrenergic receptors in goldfish

In the present study we examined the effects of intracerebroventricular (ICV) or intraperitoneal (IP) galanin administration on food intake in satiated goldfish, at different time intervals after injections, 0-2, 2-8 and 0-8 h. We found that food intake was increased by ICV administered galanin (1 and 3.33 micrograms) at 2 and 8 h postinjection, while no modifications on feeding were observed after intraperitoneal injection in any of the studied time intervals. The galanin receptor antagonist, galantide, blocked the galanin-induced feeding increase. Pretreatment with yohimbine (alpha 2-adrenergic receptor antagonist), but not with prazosin (alpha 1-adrenergic receptor antagonist) antagonized the stimulatory effect of galanin on ingestive behavior. These results suggest that galanin is involved in the central regulation of feeding in goldfish, being the food intake stimulatory effect mediated by the alpha 2-adrenergic system.

NPY receptors and opioidergic system are involved in NPY-induced feeding in goldfish

The present study evaluated the effects of both intraperitoneal (i.p. ) and intracerebroventricular administration of selective Y(1) [(Leu(31), Pro(34))-NPY] and Y(2) [(Pro(13), Tyr(36))-NPY (13-36)] receptor agonists on food intake in satiated goldfish. Food intake (FI) was significantly increased by central administration of the Y(1) agonist (1 microg), but not by the Y(2) agonist, at 2 h postinjection. The feeding increase induced by (Leu(31), Pro(34))-NPY was in a similar magnitude to that obtained after ICV injection of the neuropeptide Y, and both feeding stimulations were reversed by the NPY (27-36), a general NPY antagonist. The i.p. administration of the agonists either did not significantly modify (Y(2) agonist) or decreased (Y(1) agonist) food intake in goldfish. These data indicate that it is the Y(1)-like (similar to Y(1) and/or Y(5)) receptor, and not Y(2), that is involved in the central modulation of the feeding behavior in goldfish. We also investigated the possible involvement of opioid peptides as mediators of the NPY stimulatory action on food intake in goldfish. The ICV administration of naloxone (10 microg), a general opioid antagonist, blocked the NPY-induced feeding in goldfish, suggesting that the opioidergic system is involved in feeding regulation by NPY.

Central administration of β-endorphin increases food intake in goldfish: pretreatment with the opioid antagonist naloxone

The effect of intraperitoneal or intracerebroventricular beta-endorphin administration on food intake has been studied in satiated goldfish. Food intake was evaluated at different time intervals after injections, 0-2, 2-8 and 0-8 h. The 0.1 and 1 micrograms doses of beta-endorphin intracerebroventricularly administered induced an increase in food intake during the first 2 h postinjection, while no modifications on feeding were observed in the next 6 h. These same doses of beta-endorphin used increased cumulative food intake at 8 h postinjection. In contrast, intraperitoneal injection of 1 micrograms of beta-endorphin did not modify food intake in any of the studied time intervals. Naloxone, an opioid antagonist, attenuated the beta-endorphin-induced feeding increase. These results suggest that opioids may play a role in modulation of feeding central regulation, acting via opioid receptors in goldfish.

Melatonin reduces body weight in goldfish (Carassius auratus): effects on metabolic resources and some feeding regulators.

Abstract:  The objective of the present study was to analyze the effects of chronic melatonin (10 μg/g body weight) on body weight and the main energetic reserves, particularly hepatic and muscle content of proteins, lipids and glycogen in goldfish. In addition, we studied plasma leptin and ghrelin, and hypothalamic content of neuropeptide Y (NPY) and monoamines after chronic melatonin treatment in order to elucidate a possible interplay between melatonin and these feeding regulators on the body weight regulation in this species. Body weight gain and specific growth rate were reduced (74% and 76%, respectively) after chronic (10 days) intraperitoneal (i.p.) treatment with melatonin. The carbohydrate and lipid metabolism was regulated by melatonin in goldfish, because this indoleamine reduced muscle glycogen stores and increased lipid mobilization. A suppressive trend, but not statistically significant, in circulating ghrelin was observed after chronic treatment with melatonin. Chronic melatonin administration significantly reduced noradrenergic metabolism and increased dihydroxiphenylacetic acid content in the hypothalamus, without significant modifications in the serotoninergic system. Thus, it could be suggested that melatonin may mediate its action on energy balance in fish, at least in part, via interactions with hypothalamic catecholaminergic system. Plasma leptin and hypothalamic NPY remained unaltered after melatonin treatment, suggesting that these feeding regulators may not be involved in the effects of melatonin on energy homeostasis in fish.

Feeding time synchronizes clock gene rhythmic expression in brain and liver of goldfish (Carassius auratus).

Little is known about the feeding time dependence of clock gene expression in fish. The aim of the present study was to investigate whether a scheduled feeding time can entrain the rhythmic expression of several clock genes (period and cryptocrome) in the brain and liver of a teleost, the goldfish. Fish maintained under continuous light (LL) conditions were divided into 3 groups. Two groups were fed daily at 1000 h and 2200 h, respectively, and the third group was subjected to a random schedule regime. After 30 days, the fishes under 24-h food deprivation were sacrificed through a 24-h cycle, and clock gene expression in the optic tectum, hypothalamus, and liver was quantified by real-time PCR. The findings pointed to differences between the central and peripheral tissues studied. In the absence of a light-dark cycle (constant light), a scheduled feeding regime was necessary and sufficient to maintain both the rhythmic expression of several clock genes in the optic tectum and hypothalamus, as well as daily rhythms in locomotor activity. In contrast, neither locomotor activity nor clock gene expression in brain tissues was synchronized in randomly fed fish. However, in the liver, most of the clock genes studied presented significant daily rhythms in phase (related to the time of the last meal) in all 3 experimental groups, suggesting that the daily rhythm of clock genes in this organ only depends on the last meal time. The data suggest that, as in mammals, the smooth running of the food entrainable oscillator (FEO) in fish involves the rhythmic expression of several clock genes (Per1 and Cry3) in the central and peripheral structures. The results also indicate that the food anticipatory activity (FAA) in goldfish is not only the result of rhythmic clock gene expression in the liver because rhythmic clock gene expression was observed in randomly fed fishes, while FAA was not observed.

Mu-opioid receptor is involved in β-endorphin-induced feeding in goldfish

The present study evaluated the central effects of selective opioid receptor subtype agonists and antagonists on food intake in satiated goldfish. Significant increases in feeding behavior occurred in goldfish injected with beta-endorphin, the kappa agonist, U-50488, the delta agonist, [D-Pen2,D-Pen5]enkephalin (DPEN), and the mu agonist, [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin (DAMGO). On the other hand, the different receptor antagonists used: nor-binaltorphamine (nor-BNI) for kappa, 7-benzidilidenenaltrexone (BNTX) for delta 1, naltriben for delta 2, beta-funaltrexamine (beta-FNA) for mu, and naloxonazine for mu 1, by themselves, did not modify ingestion or slightly reduced it. The feeding stimulation by beta-endorphin was antagonized by beta-FNA and naloxonazine, but not by nor-BNI, BNTX, or naltriben. These data indicate that the mu-opioid receptor is involved in the modulation of the feeding behavior in goldfish.