Neuronal relationship between orexin-A- and neuropeptide Y-induced orexigenic actions in goldfish

Orexin-induced orexigenic action is mediated by neuropeptide Y (NPY) in goldfish and rodents. A previous study indicated that NPY-induced orexigenic action may also be mediated by orexin-A in goldfish. However, there is little information about the mutual actions of orexin-A and NPY in the goldfish. Therefore, using their specific receptor antagonists, we examined whether the orexigenic actions of orexin-A and NPY mutually interact in the goldfish. The stimulatory effect of intracerebroventricular injection of NPY at 1 pmol/g body weight (BW) on food intake was abolished by treatment with the orexin receptor-1 antagonist, SB334867, at 10 pmol/g BW whereas the NPY Y1-receptor antagonist, BIBP3226, at 100 pmol/g BW attenuated orexin-A (at 2.8 pmol/g BW)-stimulated feeding. This led us, using a double-immunostaining method and confocal laser scanning microscopy, to investigate whether orexin-A- and NPY-containing neurons in the goldfish brain have direct mutual inputs. Orexin-A- and NPY-like immunoreactivities were distributed throughout the brain, especially in the diencephalon. Orexin-A- and NPY-containing neurons were located in a region of the hypothalamus, the nucleus posterioris periventricularis (NPPv), in close proximity to each other: NPY-containing nerve fibers or endings lay in close apposition to orexin-A-containing neurons in the NPPv, and orexin-A-containing nerve fibers or endings also lay in close apposition to NPY-containing neurons in the same region. These results indicate that, in goldfish, orexin-A- and NPY-induced orexigenic actions are mediated by mutual signaling pathways.     

Neuropeptide Y (NPY) May Integrate Responses of Hypothalamic Feeding Systems and the Hypothalamo-Pituitary-Adrenal Axis

Neuropeptide Y (NPY) is a powerful stimulus to food intake in the rat. Exogenous NPY given into the third ventricle or into the paraventricular nucleus (PVN) of the hypothalamus stimulates both food consumption as well as the hypothalamus-pituitary-adrenal (HPA) axis. Presumably NPY activates the adrenocortical system through direct stimulation of CRF containing cells in the PVN. Food intake is also a major regulator of adrenocortical activation. Rhythms in HPA axis activity follow rhythms in food consumption, and rats that have been food deprived overnight have inhibited HPA axis responses to restraint stress and corticosteroid feedback the following morning. To investigate the interaction of NPY with both feeding and HPA axis activation three sets of experiments were performed: Animals fed ad lib were injected icv with NPY (2.5 μg) and allowed access to food or not post injection; animals were fasted overnight prior to NPY injection; finally, dose response experiments were performed to examine the relative sensitivities of feeding and HPA axis activation to exogenous NPY. Ad lib fed animals allowed access to food after NPY injection had slightly greater ACTH responses to NPY while glucocorticoid and insulin responses were not significantly different from ad lib fed animals not allowed access to food post injection. Animals allowed to eat post injection had significantly decreased food consumption the night following injection, however, total 24 h food consumption was not different between these animals and those given food 8 h post NPY injection. In overnight fasted animals NPY injections produced ACTH responses of equal magnitude to those in ad lib fed animals. Insulin responses to NPY were significantly elevated compared to CSF controls in overnight fasted animals. Dose response studies revealed that the adrenocortical system responds to icv NPY with at least as great sensitivity as feeding systems. NPY is discussed as a potential integrator of feeding and responsiveness in the HPA axis.     

Neuropeptide Y and food intake in fasted rats: effect of naloxone and site of action

Central administration of neuropeptide Y (NPY) induces food intake in freely feeding animals and this effect is mediated by hypothalamic sites. Little is known, however, about the effect of NPY on food intake and site of action in food-deprived animals. To examine this further, 24-h fasted rats received injections of saline or NPY into the lateral cerebral ventricle (10 micrograms/10 microliters; n = 8) or into the lateral (LH) or ventromedial hypothalamus (VMH) (1 microgram/0.5 microliters; n = 44). In addition, intracerebroventricular (i.c.v.) injections of NPY were carried out with or without i.c.v. naloxone (25 micrograms), a specific opioid receptor antagonist. During the first 40 min food intake was not different with or without NPY. After 60 and 120 min, food intake was 5.9 +/- 0.4 g and 8.3 +/- 0.6 g with i.c.v. saline which was significantly augmented by i.c.v. NPY to 8.7 +/- 0.9 g and 14.4 +/- 1.5 g, respectively (P less than 0.05). This increase in food consumption was due to a prolongation of feeding time. The opioid receptor antagonist naloxone significantly augmented latency to feed, both in the absence and presence of NPY (8.0 vs 1.7 min or 14.7 vs 2.8 min, respectively) and abolished the NPY-induced increase in food intake. Following intrahypothalamic injection of NPY, an increase in food intake (greater than 20%) was observed in 50% of the histologically identified LH and VMH sites, but only in 15% of the injection sites outside the LH/VMH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Role of leptin in the control of feeding of goldfish Carassius auratus: interactions with cholecystokinin,neuropeptide Y and orexin A,and modulation by fasting

To assess the role of leptin on food intake regulation in goldfish, we examined the effects of central (intracerebroventricular, ICV) and peripheral (intraperitoneal, IP) injections of recombinant murine leptin on feeding behavior. Centrally (100 ng/g) and peripherally (300 ng/g) injected leptin both caused a significant decrease in food intake, compared to the saline-treated controls. To test the hypothesis that leptin influenced orexigenic neuropeptide systems in goldfish, fish were co-injected with neuropeptide Y (NPY) or orexin A and leptin. Both NPY (5 ng/g) and orexin A (10 ng/g) significantly increased food intake. Fish co-injected ICV with NPY (5 ng/g) or orexin A (10 ng/g) and leptin (1 or 10 ng/g) had a food intake lower than that of fish treated with NPY or orexin A alone. NPY mRNA expression in goldfish brain was reduced 2 and 6 h following central injection of leptin. To test the hypothesis that the cholecystokinin (CCK) mediates the effects of leptin in goldfish, fish were simultaneously injected ICV with an ineffective dose of leptin (10 ng/g) and either ICV or IP with an ineffective doses of CCK (1 ng/g ICV or 25 ng/g IP). These fish had a food intake lower than vehicle-treated fish, suggesting that leptin potentiates the satiety actions of CCK. CCK hypothalamic mRNA expression was increased 2 h following central treatment with leptin. The CCK receptor antagonist proglumide blocked both central and peripheral CCK satiety effects. Blockade of CCK brain receptors by proglumide resulted in an inhibition of the leptin-induced decrease in food intake and an attenuation of the inhibiting action of leptin on both NPY- and orexin A-induced feeding. These data suggests that CCK has a role in mediating the effects of leptin on food intake. Fasting potentiated the actions of leptin and attenuated the effects of CCK. Whereas fasting had no effects on the brain mRNA expression of CCK, it increased the brain mRNA expression of NPY and decreased the expression of CART. These changes in neuropeptide expression were partially reversed when fish were treated ICV with leptin. These results provide strong evidence that, in goldfish, leptin influences food intake, in part by modulating the orexigenic effects of NPY and orexin and that its actions are mediated, at least in part, by CCK.     

Regulation by orexin of feeding behaviour and locomotor activity in the goldfish

Orexin is a hypothalamic neuropeptide that is implicated in the regulation of feeding behaviour and the sleep-wakefulness cycle in mammals. However, in spite of a growing body of knowledge concerning orexin in mammals, the orexin system and its function have not been well studied in lower vertebrates. In the present study, we first examined the effect of feeding status on the orexin-like immunoreactivity (orexin-LI) and the expression of orexin mRNA in the goldfish brain. The number of cells showing orexin-LI in the hypothalamus of goldfish brain showed a significant increase in fasted fish and a significant decrease in glucose-injected fish. The expression level of orexin mRNA in the brains of fasted fish increased compared to that of fed fish. We also examined the effect of an i.c.v. injection of orexin or an anti-orexin serum on food intake and locomotor activity in the goldfish. Administration of orexin by i.c.v. injection induced a significant increase of food intake and locomotor activity, whereas i.p. injection of glucose or i.c.v. injection of anti-orexin serum decreased food consumption. These results indicate that the orexin functions as an orexigenic factor in the goldfish brain.     

NPY mediates the feeding elicited by muscimol injections into the nucleus accumbens shell

Injections of muscimol into the nucleus accumbens shell (AcbSh) induce large increases in food intake in satiated rats and also activate neurons in a number of feeding-related brain regions, including NPY-containing neurons in the arcuate hypothalamic nucleus and cells in the paraventricular hypothalamic nucleus. This suggests that the NPY system may participate in the expression of AcbSh-mediated feeding behavior. Therefore, we examined the effects of intraventricular administration of the Y1 receptor antagonist 1229U91 or the Y5 receptor antagonist L-152,804 on AcbSh-mediated food intake. Intra-AcbSh muscimol elicited a large increase in food intake which was potently suppressed by blocking either central Y1 or Y5 receptors. Our results suggest that the AcbSh influences food intake, in part, through the release of NPY.     

Involvement of hypothalamic neuropeptide Y in pentazocine induced suppression of food intake in rats

The potent orexigenic peptide neuropeptide Y (NPY) has been considered as a possible endogenous ligand for a subpopulation of sigma receptors (SigR). However, their mutual interaction with reference to feeding behavior remains poorly understood. In the present study, we explored the possible interaction between sigma1 receptors (Sig1R) agonist, pentazocine, and NPY on food intake in satiated rats. While pentazocine dose-dependently reduced the food intake, NPY significantly increased it at 2, 4 and 6 h post injection time points. In combination studies, pretreatment with NPY (0.1 nmol/rat, intra-PVN) normalized the inhibitory effect of pentazocine (60 μg/rat, intra-PVN) on food intake. Similarly, pre-treatment with pentazocine (30 μg/rat, intra-PVN) significantly antagonized the orexigenic effect of NPY (0.5 and 1.0 nmol/rat, intra-PVN). Moreover, pentazocine treatment decreased NPY immunoreactivity in arcuate (ARC), paraventricular (PVN), dorsomedial (DMH) and ventromedial (VMH) nuclei of hypothalamus. However, no change was observed in lateral hypothalamus (LH). Study implicates the reduced NPY immunoreactivity for the anorectic effect observed following pentazocine injections. Therefore, the concomitant activation of the NPYergic system along with the Sig1R agonist treatment may serve a useful purpose in the management of the unwanted side effects related to energy homeostasis.     

Paraventricular nucleus injections of naloxone methiodide inhibit NPY's effects on energy substrate utilization

Microinjection of neuropeptide Y (NPY) into the paraventricular nucleus (PVN) of the hypothalamus stimulates eating and increases respiratory quotient. In contrast, administration of opioid receptor antagonists reduces food intake and suppresses NPY-induced feeding. The present study examined whether naloxone methiodide, an opioid antagonist, would suppress the potentiation of NPY on energy substrate utilization, when injected into the PVN. Naloxone methiodide was injected at doses of 0.1 and 1.0 g, 10 min prior to NPY treatment. NPY was administered immediately prior to the start of the nocturnal period and RQ was determined using an open-circuit calorimeter. Doses of 50 and 100 pmol NPY alone evoked reliable increases in RQ within 30min of treatment. Following naloxone methiodide pretreatment, the stimulatory action of NPY was significantly attenuated. These data indicate that opioid receptors in the PVN influence the action of NPY on energy substrate utilization.     

Orexin-induced food intake involves neuropeptide Y pathway

Orexins (orexin-A and -B) are recently identified neuropeptides, which are thought to be implicated in the regulation of feeding behavior. We used a NPY-Y1 receptor specific antagonist, BIBO3304, to examine whether NPY is involved in orexin-induced feeding behavior. Intracerebroventricular administration of orexin-A (10 nmol) induced food intake in rats (food intake for 3 h; vehicle 0.3+/-0.2 g vs. orexin-A 10 nmol, 4.0+/-0.5 g, n=4). Orexin-induced feeding behavior was partially inhibited by prior administration of BIBO3304 (3 h food intake: orexin-A 10 nmol, 4.0+/-0.5 g vs. BIBO3304 (60 microgram) + orexin-A 10 nmol, 2.2+/-0.2 g, n=4). A low dose of BIBO3304 (30 microgram) did not show a significant inhibitory effect. BIBO3457, an inactive enantiomer, used as a negative control, did not show any inhibitory effect on orexin-A-induced feeding behavior. Fos expression was observed in NPY-containing neurons in the arcuate nucleus 1 h after orexin-A (10 nmol) was administered intracerebroventricularly (control 0.3+/-0.08%, orexin-A 10.2+/-0.8%, n=5 rats/group). These observations suggest that NPY is involved in orexin-induced feeding behavior. However, BIBO3304 did not completely abolish the effect of orexin-A. These results suggest that orexin-A elicits feeding behavior partially via the NPY pathway. The NPY system could be the one of downstream pathways by which orexin-A induces feeding behavior. Another pathway may also be involved in orexin-A-induced feeding behavior, because BIBO3304 did not completely abolish orexin-A-induced feeding behavior.     

Neuropeptide Y (NDY) Y1 Receptoe mRNA is Upregulated in Association with Transient Hyperphagia and Body Weight Gain: Evidence for a Hypothalamic Site for Concurrent Development of Leptin Resistance

Microinjection of colchicine (COL), a neurotoxin that blocks axoplasmic flow in the neurons, bilaterally into the ventromedial nucleus (VMN) evokes transient hyperphagia and body weight gain. These shifts in energy balance occurred in conjunction with development of increased sensitivity to neuropeptide Y (NPY), the endogenous orexigenic signal. In order to trace the aetiology of NPY supersensitivity, we have evaluated (1) NPY Y1 and Y5 receptor (R) gene expression in the hypothalamus and (2) the possibility of alterations in the inhibitory action of leptin, a hormone produced by lipocytes. Adult male rats were rendered hyperphagic with bilateral microinjections of COL (4 μg/side) into the VMN. We observed that hypothalamic NPY Y1 mRNA levels, as measured by RNAase protection assay, were significantly increased on day 2 and returned to the control level on day 4 in COL-injected rats. The effects on NPY Y5R mRNA were not as clear cut. Interestingly, serum leptin levels increased in association with the hyperphagia and body weight gain, thereby raising the likelihood of development of resistance to the suppressive effect of endogenous leptin on food intake. Indeed, intracerebroventricular injection of 7 μg human recombinant leptin, a dose that attenuated daily food intake in normal and fasted rats, was completely ineffective in attenuating hyperphagia in COL-treated rats. These results show that transient hyperphagia induced by interruption of signalling in the VMN may be caused by increased sensitivity to NPY, which may be caused, in part, by increased expression of NPY Y1R in hypothalamic sites involved in regulation of ingestive behaviour. Additionally, the observation of increased leptin release and concurrent development of leptin resistance suggest that a normally functioning VMN may be necessary for the central inhibitory effects of leptin on food intake.     

Neuropeptide tyrosine-like immunoreactive system in the brain,olfactory organ and retina of the zebrafish,Danio rerio,during development

The anatomical distribution of neuropeptide tyrosine (NPY)-like immunoreactivity was investigated in the brain, olfactory organ and retina of the zebrafish, Danio rerio, during development and in juvenile specimens, by using the indirect immunofluorescence and the peroxidase-antiperoxidase methods. In 60 h post fertilization (hpf) embryos, NPY-like immunoreactive cell bodies appeared in the hypothalamus, within the posterior periventricular nucleus. Few positive nerve fibers were found in the hypothalamus and in the tegmentum of the mesencephalon. In 72 hpf embryos, a new group of NPY-like immunoreactive cells was found in the olfactory pit. At day 4 of development, NPY-like immunoreactive cell bodies were detected between the olfactory pit and the olfactory organ. In the hypothalamus the location of positive cell bodies was similar to that reported in the previous developmental stages. A few positive nerve fibers appeared in the tegmentum of the rhombencephalon. At days 7 and 15 of development, the distribution of NPY-like immunoreactivity was very similar to that reported at day 4. However, at day 15, NPY-like immunoreactivity appeared for the first time in amacrine cells of the retina and in nerve fibers of the tectum of the mesencephalon. In 1-month/3-month-old animals, additional groups of NPY-like immunoreactive cell bodies appeared in the glomerular layer of the olfactory bulbs, the terminal nerve, the lateral nucleus of the ventral telencephalic area, the entopeduncular nucleus and in the medial region of the reticular formation of the rhombencephalon. These results show that NPY-like immunoreactive structures appear early during ontogeny of zebrafish. The distribution of the immunoreactive system increases during the ontogeny, the juvenile stages, and reaches the complete development in mature animals. The location of NPY-like immunoreactivity indicates that, during development, NPY could be involved in several neuromodulatory functions, including the processing of visual and olfactory information. In 1-month/3-month-old animals, NPY-like immunoreactive nerve fibers are present in the pituitary, suggesting that, from these stages onward, NPY may influence the secretion of pituitary hormones.     

Anorectic Effect of Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) in Rats: Lack of Evidence for Involvement of Hypothalamic Neuropeptide Gene Expression

We investigated the effect of centrally administered pituitary adenylate cyclase activating polypeptide (PACAP) on feeding in rats, and the involvement of hypothalamic neuropeptide gene expression using in situ hybridization. lntracerebroventricular injection of PACAP (1000  pmol/rat) significantly decreased food intake in a dose-dependent manner. In PACAP-treated rats, neuropeptide Y (NPY) mRNA levels in the arcuate nucleus and galanin mRNA levels in the paraventricular nucleus increased, and corticotropin-releasing hormone (CRH) mRNA levels in the paraventricular nucleus decreased. In rats fasted for 72  h, NPY mRNA levels increased, and CRH mRNA levels decreased, but galanin mRNA levels were unchanged. These results indicate that the anorectic function of PACAP is not mediated by NPY or CRH, and that PACAP increases galanin synthesis.     

Hypothalamic paraventricular nucleus injections of urocortin alter food intake and respiratory quotient

Corticotropin releasing hormone (CRH) acts on the central nervous system to alter energy balance and influence both food intake and sympathetically-mediated thermogenesis. CRH is also reported to inhibit food intake in several models of hyperphagia including neuropeptide Y (NPY)-induced eating. The recently identified CRH-related peptide, urocortin (UCN), also binds with high affinity to CRH receptor subtypes and decreases food intake in food-deprived and non-deprived rats. The present experiment characterized further the feeding and metabolic effects of UCN by examining its impact after direct injections into the paraventricular nucleus (PVN) of the hypothalamus. In feeding tests (n=8), UCN (50-200 pmol) was injected into the PVN at the onset of the dark cycle and food intake was measured 1, 2 and 4 h postinjection. In separate rats (n=8), the metabolic effects of UCN were monitored using an open circuit calorimeter which measured oxygen consumption (V(O2)) and carbon dioxide production (V(CO2)). Respiratory quotient (RQ) was calculated as V(CO2)/V(O2). UCN suppressed feeding at all times studied and reliably decreased RQ within 30 min of infusion. Additional work examined the effect of UCN (50-100 pmol) pretreatment on the feeding and metabolic effects of NPY. NPY, injected at the start of the dark period, reliably increased 2 h food intake. This effect was blocked by PVN UCN administration. Similarly, UCN blocked the increase in RQ elicited by NPY alone. These results suggest that UCN-sensitive mechanisms within the PVN may modulate food intake and energy substrate utilization, possibly through an interaction with hypothalamic NPY.     

Metabolic and Orexigenic Effects of Intracerebroventricular Neuropeptide Y are Attenuated by Food Deprivation

Administration of neuropeptide Y (NPY) into the hypothalamus or cerebral ventricles has been shown to increase food intake, the secretion of hormones such as insulin, glucagon and corticosterone and to alter the metabolism of carbohydrate and lipids. It has been suggested that metabolic effects of hypothalamic NPY may contribute to fat accretion in some types of obesity and to the metabolic and behavioural adaptation to food deprivation. However, it is currently unknown if different nutritional states alter the responses to hypothalamic NPY. Consequently, we have compared the effects of NPY injected into the third ventricle (ICV) in the fed and overnight-fasted state on ingestive behaviour, on insulin, glucagon and corticosterone secretion before, and following, an IV glucose bolus (IVGTT) and on blood glucose following an intra-arterial insulin bolus (ITT). Studies were performed on conscious, unrestrained adult female rats. In the fed state, 2 and 6  &mgr;g ICV NPY produced a potent orexigenic and dypsogenic effect. In the fasted state, the 2  &mgr;g dose had a dypsogenic effect, while only the 6  &mgr;g dose had a significant orexigenic effect. In the fed but not fasted state, 3  &mgr;g ICV NPY increased plasma glucagon and corticosterone levels and attenuated the decline in blood glucose during the ITT. By contrast, in both fed and fasted groups, 3  &mgr;g ICV NPY potentiated the insulin secretory response during the IVGTT.     

Neuropeptide-Y: a possible mediator of prolactin-induced feeding and regulator of energy balance in the ring dove (Streptopelia risoria)

Although neuropeptide-Y (NPY) has been widely reported to be a potent stimulator of feeding activity and regulator of energy homeostasis, most of the supportive evidence for such effects has been gathered in mammalian species. This study characterized the orexigenic potency of NPY in an avian species, the ring dove, and measured changes in hypothalamic NPY-immunoreactive (NPY-ir) cell numbers in response to energy state fluctuations or intracranial administration of the potent orexigenic hormone prolactin. Food intake was significantly elevated in male doves at 1 h after intracerebroventricular (i.c.v.) injection of 0.25 and 0.5 microg NPY but not after injection of a higher dose (1.0 microg). In time course studies, food intake was increased at 1 h after i.c.v. injection of 0.5 microg NPY but was not elevated at 2, 3, or 4 h. The number of NPY-ir cell bodies in the infundibular region of the dove hypothalamus increased two to four-fold following acute food deprivation, chronic food restriction, or repeated i.c.v. injections of prolactin. No additive effects were observed when food restriction and prolactin treatment were combined. These findings suggest that NPY is involved in energy homeostasis in doves and are consistent with the hypothesis that prolactin-induced hyperphagia is mediated in part by NPY.     

Nesfatin-1 suppresses energy intake, co-localises ghrelin in the brain and gut, and alters ghrelin, cholecystokinin and orexin mRNA expression in goldfish

Nesfatin‐1 is a novel anorectic peptide encoded in the precursor protein nucleobindin‐2 (NUCB2). We recently reported the presence and appetite suppressing effects of nesfatin‐1 in goldfish. Nesfatin‐1 has been co‐localised with ghrelin in the stomach of rats. Whether nesfatin‐1 influences other appetite regulatory peptides in goldfish remains unclear. The main objectives of the present study were to investigate whether nesfatin‐1 co‐localises ghrelin in goldfish, and to test whether exogenous nesfatin‐1 influences endogenous ghrelin, cholecystokinin (CCK) and orexin A (OXA). We found co‐localisation of nesfatin‐1‐like and ghrelin‐like immunoreactivity in the enteroendocrine cells of the goldfish anterior intestine (J‐loop). Furthermore, co‐localisation of ghrelin and nesfatin‐1 was also observed in the posterior nucleus lateralis tuberis of the goldfish hypothalamus, a brain region implicated in the regulation of food intake. These findings suggest a functional relationship between ghrelin and nesfatin‐1 in goldfish. In support of this, i.c.v. administration of goldfish (gf) nesfatin‐1 [25 ng/g body weight (BW)], suppressed food intake and the expression of mRNAs encoding preproghrelin, ghrelin receptor (GHS‐R 1a‐1), CCK and NUCB2 in the forebrain of fed fish, as well as ghrelin and NUCB2 mRNA in the hypothalamus of unfed fish, both at 1 h post‐injection. Nesfatin‐1 stimulated hypothalamic CCK mRNA expression at 30 min post‐injection in fed fish, and inhibited OXA mRNA in the unfed fish hypothalamus 1 h post‐injection. Similarly, i.c.v. injections of gfghrelin (1 ng/g BW), although stimulating food intake, suppressed NUCB2 and preproghrelin mRNAs, but not ghrelin receptor mRNA expression in the forebrain. It is also evident that exogenous ghrelin and nesfatin‐1 mRNAs encoding these peptides. Our novel results indicate interactions between nesfatin‐1 and ghrelin, CCK and orexin, and show that nesfatin‐1 acts on other appetite regulatory peptides in a time‐ and feeding status‐dependent, as well as tissue‐specific, manner in goldfish.     

Effect of peripheral 2-DG on opioid and neuropeptide Y gene expression

It is well known that 2-Deoxy-d-glucose (2-DG) blocks intracellular utilization of glucose and increases food intake. The aim of the present study was to determine whether administration of 2-DG alters gene expression of the orexigenic peptides, neuropeptide Y (NPY) and endogenous opioids, in the arcuate nucleus of the hypothalamus (ARC). Male Sprague–Dawley rats were injected peripherally (i.p.) with 2-DG (200 or 400 mg/kg body weight) and were sacrificed at 2 or 6 h post injection. Half of the animals were given ad libitum access to food whereas the other half of the animals were food-deprived. 2-DG increased food intake fourfold compared to saline injected animals, but did not affect NPY mRNA levels after 2 h. Messenger RNA levels of ProDynorphin (proDYN), but not pro-opiomelanocortin (POMC) nor proEnkephalin (proENK) were significantly decreased 2 h after 2-DG injection. Administration of 400 mg/kg of 2-DG increased mRNA levels of NPY in the arcuate nucleus after six h, but only in those animals not receiving food.     

Involvement of specific orexigenic neuropeptides in sweetener-induced overconsumption in rats

Palatability is one of the factors that regulates food and fluid intake and contributes to overconsumption in turn contributing to obesity. To elucidate the brain mechanisms of the palatability-induced ingestion, we explored the roles of six hypothalamic orexigenic neuropeptides, orexin, melanin-concentrating hormone (MCH), neuropeptide Y (NPY), agouti-related protein (AgRP), ghrelin and dynorphin, in the intake of a palatable solution, saccharin. Of the six peptides, intracerebroventricular (i.c.v.) administrations of orexin, MCH and NPY increased the intake of saccharin. Drinking of saccharin in turn elevated the mRNA levels of orexin and NPY, but not MCH. Pre-treatments of naloxone, an opioid antagonist, blocked the orexigenic effects of orexin and NPY. Specific gastric motor responses induced by central orexin-A and NPY are well known, however, MCH did not induce such responses. The i.c.v. administration of orexin-A facilitated gastric emptying. These results suggest that the overconsumption promoted by sweet and palatable tastes is attributed to the activation of orexigenic neuropeptides, such as orexin and NPY, and a downstream opioid system together with enhanced digestive functions.