Hypothalamic Orexigenic Neuropeptide Research Articles

Effect of CCK-8 on insulin-induced hyperphagia and hypothalamic orexigenic neuropeptide expression in the rat

The influence of cholecystokinin octapeptide (CCK-8) on normal and insulin-induced feeding and expression of orexigenic hypothalamic neuropeptides was investigated in male rats. CCK-8, administered during meals (4 μg/kg) or continuously (32 μg/kg over 60 min), blunted the stimulating effect of insulin (50 IU/kg) on feeding by reducing meal size (−60%; P < 0.05 or −86%; P < 0.0001, respectively). Rats without access to food and injected with IP insulin (50 IU/kg) showed increased hypothalamic mRNA levels of orexin (+30%; P < 0.05) and melanin-concentrating hormone (+52%; P < 0.05), as compared with ad libitum-fed and saline-injected control rats. Continuous IP infusion of CCK-8 (32 μg/kg) blunted these increases. Our results suggest that both orexin and melanin-concentrating hormone participate in the response to insulin hypoglycemia without food being present; these neurons may be involved in mechanisms related to insulin-induced hyperphagia. Signals triggered by peripheral CCK-8 act to decrease the expression of orexin and melanin-concentrating hormone. This may be associated with a reduction in hyperphagia.

Genetic deletion of ghrelin does not decrease food intake but influences metabolic fuel preference.

Ghrelin is a recently identified growth hormone (GH) secretogogue whose administration not only induces GH release but also stimulates food intake, increases adiposity, and reduces fat utilization in mice. The effect on food intake appears to be independent of GH release and instead due to direct activation of orexigenic neurons in the arcuate nucleus of the hypothalamus. The effects of ghrelin administration on food intake have led to the suggestion that inhibitors of endogenous ghrelin could be useful in curbing appetite and combating obesity. To further study the role of endogenous ghrelin in appetite and body weight regulation, we generated ghrelin-deficient (ghrl(-/-)) mice, in which the ghrelin gene was precisely replaced with a lacZ reporter gene. ghrl(-/-) mice were viable and exhibited normal growth rates as well as normal spontaneous food intake patterns, normal basal levels of hypothalamic orexigenic and anorexigenic neuropeptides, and no impairment of reflexive hyperphagia after fasting. These results indicate that endogenous ghrelin is not an essential regulator of food intake and has, at most, a redundant role in the regulation of appetite. However, analyses of ghrl(-/-) mice demonstrate that endogenous ghrelin plays a prominent role in determining the type of metabolic substrate (i.e., fat vs. carbohydrate) that is used for maintenance of energy balance, particularly under conditions of high fat intake.

Hypothalamic malonyl-CoA as a mediator of feeding behavior.

Previous studies showed that i.p. administration of C75, a potent inhibitor of fatty acid synthase (FAS), blocked fasting-induced up-regulation of orexigenic neuropeptides and down-regulation of anorexigenic neuropeptides in the hypothalami of mice. As a result, food intake and body weight were drastically reduced. Here we provide evidence supporting the hypothesis that hypothalamic malonyl-CoA, a substrate of FAS, is an indicator of global energy status and mediates the feeding behavior of mice. We use a sensitive recycling assay to quantify malonyl-CoA to show that the hypothalamic malonyl-CoA level is low in fasted mice and rapidly (< or = 2 h) increases (approximately 5-fold) on refeeding. Intracerebroventricular (i.c.v.) administration of C75 to fasted mice rapidly (< or = 2 h) increased (by 4-fold) hypothalamic malonyl-CoA and blocked feeding when the mice were presented with food. Moreover, prior i.c.v. administration of an acetyl-CoA carboxylase inhibitor, 5-(tetradecyloxy)-2-furoic acid, rapidly (although only partially) prevented the C75-induced rise of hypothalamic malonyl-CoA and prevented the C75-induced decrease of food intake. These effects correlated closely with the rapid (< or = 2 h) and reciprocal effects of i.c.v. C75 on the expression of hypothalamic orexigenic (NPY and AgRP) and anorexigenic (proopiomelanocortin) neuropeptide mRNAs. Previous results showing that C75 administered i.c.v. rapidly activates hypothalamic neurons of the arcuate and paraventricular nuclei are consistent with the results reported in this paper. Together these findings suggest that level of hypothalamic malonyl-CoA, which depends on the relative activities of acetyl-CoA carboxylase and FAS, is an indicator of energy status and mediates feeding behavior.

Effect of the anorectic fatty acid synthase inhibitor C75 on neuronal activity in the hypothalamus and brainstem.

Intraperitoneal (i.p.) injection of C75, a fatty acid synthase inhibitor, causes a rapid (<or=2-h) and persistent (to at least 24-h) approximately 95% decrease in food intake. The persistent effect seems to be due to inhibition of the fasting-induced up-regulation of expression of hypothalamic orexigenic neuropeptides neuropeptide Y and agouti-related protein and down-regulation of expression of anorexigenic neuropeptides pro-opiomelanocortinalpha-melanocyte-stimulating hormone and cocaine-amphetamine-related transcript. The effect of C75 on neuronal activity in the hypothalamus and brainstem was assessed by c-Fos expression. Consistent with its effect on neuropeptide expression, C75 blocked fasting-induced c-Fos expression in the arcuate nucleus (Arc), lateral hypothalamic area (LHA), and paraventricular nucleus (PVN) 10-24 h after i.p. injection. However, i.p. C75 induced a rapid (<or=2-h) c-Fos expression in the nucleus of the solitary tract (NTS) and area postrema of the brainstem but not in the Arc or LHA. Intracerebroventricular administration of C75 rapidly induced c-Fos expression in the Arc, PVN, and NTS, supporting a central role of C75 in the regulation of food intake. Thus, suppression of food intake by C75 administered i.p. seems to be mediated in two phases, a rapid initial phase via the NTSarea postrema of the brainstem and a delayed phase via the Arc, LHA, and PVN of the hypothalamus. The delayed effect of C75 on the Arc, LHA, and PVN correlates well with its ability to interfere with the fasting-induced effects on the expression of key orexigenic (neuropeptide Y and agouti-related protein) and anorexigenic (pro-opiomelanocortinalpha-melanocyte-stimulating hormone and cocaine-amphetamine-related transcript) messages in the hypothalamus.

Antidepressant, anxiolytic and anorectic effects of a melanin-concentrating hormone-1 receptor antagonist

Melanin concentrating hormone (MCH) is an orexigenic hypothalamic neuropeptide, which plays an important role in the complex regulation of energy balance and body weight. Here we show that SNAP-7941, a selective, high-affinity MCH1 receptor (MCH1-R) antagonist, inhibited food intake stimulated by central administration of MCH, reduced consumption of palatable food, and, after chronic administration to rats with diet-induced obesity, resulted in a marked, sustained decrease in body weight. In addition, after mapping the binding sites for [(3)H]SNAP-7941 in rat brain, we evaluated its effects in a series of behavioral models. SNAP-7941 produced effects similar to clinically used antidepressants and anxiolytics in three animal models of depression/anxiety: the rat forced-swim test, rat social interaction and guinea pig maternal-separation vocalization tests. Given these observations, an MCH1-R antagonist may be useful not only in the management of obesity but also as a treatment for depression and/or anxiety.

Differential effects of a centrally acting fatty acid synthase inhibitor in lean and obese mice.

C75 is a potent inhibitor of fatty acid synthase that acts centrally to reduce food intake and body weight in mice; a single dose causes a rapid (>90%) decrease of food intake. These effects are associated with inhibition of fasting-induced up-regulation and down-regulation, respectively, of the expression of orexigenic (NPY and AgRP) and anorexigenic (POMC and CART) neuropeptide messages in the hypothalamus. Repeated administration of C75 at a submaximal level, however, differentially affected food intake of lean and obese mice. With lean mice, C75 suppressed food intake by approximately 50% and, with obese mice (ob/ob and dietary-induced obesity), by 85-95% during the first day of treatment. Lean mice, however, became tolerant/resistant to C75 over the next 2-5 days of treatment, with food intake returning to near normal and rebound hyperphagia occurring on cessation of treatment. In contrast, ob/ob obese mice responded to C75 with a >90% suppression of food intake throughout the same period with incipient tolerance becoming evident only after substantial weight loss had occurred. Dietary-induced obese mice exhibited intermediate behavior. In all cases, a substantial loss of body weight resulted. Pair-fed controls lost 24-50% less body weight than C75-treated mice, indicating that, in addition to suppressing food intake, C75 may increase energy expenditure. The decrease in body weight by ob/ob mice was due primarily to loss of body fat. In contrast to the short-term effects of C75 on "fasting-induced" changes of hypothalamic orexigenic and anorexigenic neuropeptide mRNAs, repeated administration of C75 either had the inverse or no effect as tolerance developed.

Acute orexin effects on insulin secretion in the rat: in vivo and in vitro studies

Orexin-A and orexin-B are members of a family of newly described orexigenic hypothalamic neuropeptides. Scanty data are available suggesting the involvement of orexins in regulation of the secretion of pituitary hormones and in control of energy homeostasis. Present studies aimed to explain whether orexins affect blood insulin concentration and insulin secretion in the rat. To check this possibility, adult female rats were subcutaneously injected with different doses (1 or 2 nmol) of orexin-A or orexin-B. A bolus administration of orexin-A resulted in an increase in blood insulin (up to min 120) and glucose (60 min after injection) concentration. The higher dose of orexin-B, on the other hand, exerted effect on insulin secretion only at min 60 of experiment and neither doses changed blood glucose level. Only orexin-A stimulated insulin secretion in an in vitro perfusion system of the rat pancreas preparation, while orexin-B was less effective. The results demonstrate that orexins belong to a group of neuropeptides influencing insulin secretion and acting directly on the pancreas. Direct, at least partial, effect of orexin on insulin secretion may be connected with the regulation of metabolism by this peptide.

Orexins, Orexigenic Hypothalamic Neuropeptides, Suppress the Pulsatile Secretion of Luteinizing Hormone in Ovariectomized Female Rats

It is well known that feeding disorders are deeply related to reproductive dysfunction, and some feeding regulatory factors may modulate the reproductive function. We examined the effect of orexins, the newly discovered orexigenic hypothalamic neuropeptides, on the pulsatile secretion of LH to clarify their influence on the reproductive function. We administered orexins or saline into the third ventricle of bilaterally ovariectomized (OVX) rats, and measured the serum LH concentration by RIA in blood samples drawn every 6 min for 2 hours to analyze the pulsatile secretion. In the orexin-treated groups, the mean LH concentration and the pulse frequency were significantly reduced (p < 0.01), but the pulse amplitude did not differ significantly. These data indicate that orexins suppress the pulsatile secretion of LH by influencing GnRH neurons at the hypothalamic level.

Short communication

Orexin A and orexin B are a newly described family of orexigenic hypothalamic neuropeptides. The distribution of orexin immunoreactive fibers overlaps with the luteinizing hormone-releasing hormone (LHRH) neuronal system in the septo-preoptic area and the arcuate nucleus-median eminence region. To test the hypothesis that orexins may be involved in the regulation of pituitary LH secretion by influencing LHRH release, the effects of orexin A and orexin B on LH secretion were evaluated in ovariectomized (ovx) and ovarian steroid-treated ovx rats. Intracerebroventricular injection of orexin A or orexin B rapidly stimulated LH secretion in a dose- (0.3 and 3.0 nmol) and time-related fashion in estradiol benzoate- (EB) and progesterone (P)-pretreated ovx rats. Peak plasma LH concentration at 10 min were significantly higher in orexin A- than in orexin B-injected rats. On the other hand, both orexins (3 nmol) inhibited LH release in unprimed ovx rats. This ovarian steroid-dependent bimodal LH response is reminiscent of that of other orexigenic neuropeptides, such as neuropeptide Y. These findings show that orexins belong to a group of hypothalamic messengers that neurochemically bridge the regulatory networks that controls reproduction and energy balance.

Fetal ovine obese (leptin) gene expression is regulated by circulating glucose concentrations. • 1417

Leptin, the 16 kD secreted translated product of the white adipose tissue obese (ob) gene circulates to access and suppress the adult hypothalamic orexigenic neuropeptide Y (NPY) synthesis. We have previously observed fetal hyperglycemia (↑G) and hypoglycemia (↓G) to respectively suppress or enhance fetal brain NPY synthesis (Endocrinol 1997). To determine the intermediary signal by which ↑G and ↓G regulate fetal hypothalamic NPY synthesis, we examined the role of fetal ob gene expression in this regard. Late gestation pregnant sheep were made ↑G (4.8mM; n=13) or ↓G (1.4mM; n=12) with chronic 50% glucose (G) or 5-10 μU/kg/min of insulin (I) infusions respectively and compared to euglycemic controls (2.8mM; n=12).↑G caused fetal ↑G (+80% over control) with 1-2d hyperinsulinemia (-I) followed by normoinsulinemia over 20d. ↓G led to fetal ↓G (<50% of control) with hypoinsulinemia (↓I) over 1d to 41d. Northern blots assessed fetal white adipose tissue ob gene mRNA (≈4.4 kb) concentrations with the ovine ob gene cDNA which was cloned by reverse transcription-PCR using human ob specific upstream (+56 to +75 bp) and downstream (+565 to +581 bp) primers. Sequencing of the isolated ovine ob gene cDNA revealed a predicted amino acid sequence (14 to 129 AA) with a homology of 48% to rat, 48% to mouse, 33% to human, & 39% to cow leptins. Fetal ↑G caused a two-fold increase (p< 0.05), while ↓G caused a 40% decrease in the ob gene mRNA amounts (p< 0.05). Radioimmunoassays to quantitate the circulating translated product using anti-human or anti-mouse leptin IgGs either did not or only detected≈30% respectively of the total ovine leptin. We conclude that fetal G&/or I levels reciprocally alter fetal ob gene mRNA levels. We speculate that a similar change in circulating leptin levels reciprocally affects fetal hypothalamic NPY synthesis thereby influencing postnatal food intake, body weight gain, and adiposity.