NPY Levels Research Articles

Anorexigen (TNF-alpha, cholecystokinin) and orexigen (neuropeptide Y) plasma levels in peritoneal dialysis (PD) patients: their relationship with nutritional parameters.

Malnutrition has definitely been related to mortality among dialysis patients. Persistent loss of appetite is one of the major symptoms found in these patients. It is also well recognized that several substances produce anorexia or disorders of the hunger-satiety cycle in several diseases. The aim of this study was to identify the role of anorexigen substances (TNF-alpha and cholecystokinin or CCK) and an orexigen substance (neuropeptide Y or NPY) in anorexia and malnutrition among 55 clinically stable peritoneal dialysis (PD) patients. High TNF-alpha plasma levels were found in 41 of 42 patients (97.6%) with a mean of 70.5+/-32.3 pg/ml. Patients with anorexia (n=11) or anorexia with nausea or vomiting (n=5) had higher TNF-alpha values than patients without these symptoms (75.9+/-34 vs 52.1 +/-24.5 pg/ml, P<0.05). Eight patients with a prior diagnosis of acid pylori disease showed higher TNF-alpha values (87.2+/-24.3) than 30 unaffected patients (63.6+/-30.5, P<0.05). TNF-alpha showed a significant negative linear correlation with retinol binding protein (RBP) (r=-0.37, n=34, P<0.05), and venous pH (r=-0.4, n=42, P<0.01); also, TNF-alpha values higher than 65 pg/ml were inversely associated with transferrin, cholesterol, blood urea nitrogen (BUN) and CCK. Patients with prealbumin levels lower than 30 mg/dl, a BMI lower than 30 kg/m2, nPCR lower than 1.1 g/kg/day and urea KT/V lower than 2.2 showed higher serum TNF-alpha levels. Patients who had been on CAPD treatment for longer periods showed higher TNF-alpha values. High plasma CCK levels were found in 38 of 45 patients (84%), mean 45.9+/-32.3 pg/ml. Patients with anorexia had no difference in CCK values compared with those without. A direct association was found between CCK levels and some nutritional markers (albumin, fibronectin, triglycerides, folic acid and nPCR in non diabetic patients). Although CCK has a recognized anorectic effect, this direct association might be because of an abnormal stimulation of CCK glucose feedback (trypsin) due to continuous peritoneal glucose absorption. This suggests that CCK could be an immediate food intake marker in PD patients. The NPY plasma levels were normal in 33 patients, high in 6 and low in 11. Patients with anorexia showed lower NPY levels than those without. NPY values greater than 50 pg/ml were directly associated with higher transferrin, prealbumin, RBP, nPCR and urea KT/V values. Importantly, a negative linear correlation between NPY and TNF-alpha was found (r=-0.42, n= 41, P<0.01). There was no significant relationship between residual renal clearance and the serum levels of the three peptides. In conclusion, our data suggest that high TNF-alpha and low NPY serum levels are associated with anorexia. High TNF-alpha, low CCK and low NPY serum levels are also related to a poor nutritional status. Further research on these circulating substances is required.

Effect of Intracerebroventricular Leptin Administration on Body Weight Gain in Neonatal Rats † 1581

Leptin, the secreted and translated product of the ob gene, acts upon the adult hypothalamus and decreases appetite and body weight gain. While neonatal rat and mouse adipocytic leptin mRNA and circulating peptide have been detected (FASB J 11:382-387, 1997; BBRC 238:44-47, 1997), and neonatal rat hypothalamic leptin receptors characterized (Ped Res 1998), the biological significance of leptin in the newborn remains unknown. To address this issue, 2 to 8 day old rats received intracerebroventricularly 0.5 μg (n=3) and 1μg (n=3) of murine leptin or vehicle (n=6) on a daily basis. In comparison to the vehicle group, a ≈27% body weight loss was noted in the 0.5 μg and 1 μg leptin groups by day 7 (leptin=8±0.5g vs vehicle=10.9±0.7g; p < 0.05). This weight loss persisted through day 9 (leptin=11.1±0.1g vs vehicle=15.5±0.3g; p < 0.05), declined to ≈11% by day 16 (leptin= 30.1±3g vs vehicle=36.9±0.7g; p< 0.05), and was no different at day 24. Though no change in plasma leptin or glucose concentrations was noted throughout, an increase in plasma insulin was observed only at day 3 in the 1μg leptin group (4.9±0.1 vs the vehicle group= 2.9±0.8 ng/ml; p < 0.05). In an attempt to determine the central mechanism(s) by which leptin induced this neonatal body weight loss, semi-quantitation of paraventricular neuropeptide Y immunoreactivity(NPY; aerence in NPY amounts was detected at 3d which is 24 hours following initiation of leptin administration. This is in contrast to our previous investigations where intracerebroventricular NPY led to biological changes within 24 hours (Varma et al Ped Res 41:1441, 1997). Studies assessing hypothalamic NPY levels between 7 to 24 days are in progress. We conclude that 1] exogenous intracerebroventricular leptin administration reduces body weight during the suckling phase, and 2] the anorexic/catabolic effect is observed 5 days after initiation of intracerebroventricular leptin injections. We speculate that the hypothalamic mechanism(s) for this biological response may include reduction in NPY synthesis/levels at later ages (7 to 24 d), or even perturbations in other anorexic neuro peptides(corticotropin-releasing hormone, proopiomelanocortin).

The catecholaminergic innervation of the rat amygdala.

The present study is the first to demonstrate conclusively and to analyze systematically synaptic contacts of all three types of catecholaminergic afferent fibers in different nuclei of the rat amygdala and to relate the catecholaminergic innervation to neurochemically identified target neurons. 4.1.1 Central Nucleus: The central nucleus is the amygdaloid nucleus receiving the most dense catecholaminergic innervation. In the medial central nucleus, dopaminergic, noradrenergic and adrenergic terminal plexus overlap, in the central lateral central nucleus mainly dopaminergic plexus are found. The lateral capsular central nucleus is generally scarcely innervated, but individual neurons of this subnucleus possess a dense dopaminergic innervation. Colocalization of neurotensin in dopaminergic afferents is rare, the majority of the dense neurotensin-ir terminal plexus consist of non-dopaminergic fibers. The catecholaminergic innervation of the medial central nucleus is directed preferentially at peripheral neuronal structures, and has thus presumably modulatory functions. Dopaminergic terminals form predominantly symmetric, noradrenergic and adrenergic terminals from preferentially asymmetric synapses. A characteristic feature of the dopaminergic innervation is the dense perisomatic innervation of selected neurons. Adrenergic and the majority of noradrenergic afferent fibers to the medial central nucleus originate from cell groups in the medulla oblongata and contain high levels of NPY. GAD mRNA-detection suggests that most target neurons of catecholaminergic afferent fibers are capable of synthesizing GABA in the medial central nucleus. In its dorsal part, GABA is possibly colocalized with somatostatin, and many neurons express the dopamine-1-receptor subtype mRNA. In the posteroventral medial central nucleus, on the other hand, enkephalin mRNA-r and dopamine-2-receptor subtype mRNA-reactive neurons show a similar distribution as the GAD mRNA-reactive ones. Contacts could be shown between dopaminergic, noradrenergic and adrenergic axons and NPY- and somatostatin-immunoreactive neurons which are supposedly among the brainstem projection neurons of the medial central nucleus. The dopaminergic innervation of the central lateral central nucleus resembles that of the neighboring striatum in many respects. The synaptic density is high. As in the medial subnucleus, distal neuronal elements are the preferential target structures, indicating a modulatory function possibly regulating the selectivity of the target neurons for stimuli transmitted by other afferent fibers. Besides, individual neurons possess a dense perisomatic, presumably non-selective dopaminergic innervation. The innervation does not appear to be targeted at one specific neurochemical type of neuron in the central lateral central nucleus, but rather contacts somatostatin- and neurotensin-immunoreactive neurons (which are possibly also GABAergic), in addition to GABA/enkephalin-synthesizing and other (e.g., CHAT-immunoreactive) neurons. Individual neurons of the central lateral central nucleus express the dopamine-2-receptor subtype mRNA. The dopaminergic fiber baskets of the lateral capsular central nucleus are found surrounding enkephalin mRNA-reactive neurons. Codistribution studies suggest that they express the dopamine-2-receptor subtype mRNA. 4.1.2 Basal Complex: The basal complex receives dopaminergic and noradrenergic innervation, the latter mainly originating in the locus coeruleus. Some of the dopaminergic afferents contain neurotensin, and in contrast to the central nucleus, all neurotensin-immunoreactive afferent fibers are dopaminergic. In the noradrenergic afferent fibers NPY is not detectable. These results and the innervation pattern displaying mostly peripheral neuronal target structures resemble dopaminergic and noradrenergic innervation patterns documented in cortical areas. (ABSTRACT TRUNCATED)

Immobilization stress elevates gene expression for catecholamine biosynthetic enzymes and some neuropeptides in rat sympathetic ganglia: effects of adrenocorticotropin and glucocorticoids.

Sympathetic ganglia are the major contributors to the stress-elicited rise in circulating norepinephrine, enkephalins, and neuropeptide Y. Here we examined the effect of immobilization stress and treatment with ACTH and glucocorticoids on messenger RNA (mRNA) levels for tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), preproneuropeptide Y (pre-NPY), and proenkephalin in rat superior cervical ganglia (SCG) and in stellate ganglia. Our results show a severalfold increase in the relative abundance of TH and NPY mRNAs in response to a single immobilization. Repeated stress elevated expression of all the genes studied and increased TH immunoreactivity in both ganglia. The effect of stress was more pronounced in SCG. Prolonged cortisol administration failed to alter the mRNA levels of TH, DBH, and NPY in control animals but attenuated the response to stress. In contrast, TH and DBH mRNA levels in the SCG, but not in adrenal medulla, were elevated by ACTH administration, similar to the levels attained after immobilization. The results revealed that the regulation of gene expression in response to immobilization stress in sympathetic neurons differs from the regulation in adrenal medulla. The study implicates hormonal involvement in the stress-induced changes in TH, DBH, NPY, and proenkephalin gene expression in sympathetic ganglia.

Differential Effects of the 5-HT1B/2C Receptor Agonist mCPP and the 5-HT1A Agonist Flesinoxan on Hypothalamic Neuropeptide Y in the Rat: Evidence That NPY May Mediate Serotonin's Effects on Food Intake

To determine whether NPY and 5-HT interact, we studied the effects on hypothalamic NPY and NPY mRNA levels of acute (10 mg/kg) and 7-day administration (10 mg/kg/day) of m CPP (a 5- HT 1B 2C agonist) and flesinoxan (a 5-HT 1A agonist). After both treatments, m CPP reduced food intake and NPY levels in the PVN ( p < 0.01). NPY mRNA levels were only increased by comparable food restriction ( p < 0.05). Acute flesinoxan injection increased food intake and NPY levels in the PVN and ARC ( p < 0.01). This suggests that food intake changes induced by m CPP and flesinoxan are associated with altered NPY ergic activity. Stimulation of different hypothalamic 5-HT 1 receptors may alter NPY levels available for release in the PVN, possibly by altering transport along the ARC-PVN projection. Therefore, inhibition of the ARC-PVN projection may at least partly mediate the hypophagic effects of serotonin.

Relationships between hypothalamic neuropeptide Y and food intake in the lactating rat.

hypoinsulinaemic [2]. Certain NPYergic neurones in the hypothalamic ARC that prolect to release NPY in the PVN are postulated to regulate food intake and energy balance [3-71. The ARC-PVN NPYergic projection is overactive in lactation, as shown recently by increased NPY concentrations in the ARC and PVN [8]and by increased NPY gene expression in the ARC [9,lO]. Increased NPY 1-elease in this projection is thought to drive hyperphagic? ~n lactation. The objectives of our studies were to determine if the increased hypothalamic NPY levels found during lactation are the cause or the effect of the hyperphagia, and to assess further the possible role of insulin in mediating these adaptations. Lactating rats were either freely-fed or restricted to control levels of food intake for 3 days during peak hyperphagia (13-17 days postpaltum) . To measure regional hypothalamic NPY concentrations, individual nuclei were punched out of hypothalamic sections cut on cl vibratome and assayed for NPY by RIA [Ill. Total RNA WdS extracted from whole hypothalamic blocks using the guanidium t h i o c y a n a t e p h e n o l c h l o r o f o r m method [121. RNAs were separated by gel electrophoresis and hybridized with a 'ZP-labelled rat cDNA probe. The signal intensities for NPY mRNA were normalized using the signal similarly obtained for tubulin mRNA and quantified using scanning densitometry. Plasma insulin concentrations were also determined by RIA from blood obtained at sacrifice by cardiac puncture. Compared with non-lactating controls (n=lO), freely-fed lactating rats (n=9) showed significantly increased ( p 0 . 0 5 ) NPY levels ~n the ARC and MPO (Fig. l ) , but there was no significant increase in whole hypothalamic NPY mRNA levels. Food-restricted lactating rats (n=8) showed generally higher hypothalamic NPY levels than both other groups, with significantly higher concentrations than controls (p<0.051 in the ARC, MPO, PVN and LHA (Fig. I); this group also showed NPY mRNA levels that were significantly increased (p<O. 05) beyond those of cont 1-01s. Across all three experimental groups, there was a significant inverse correlation between serum insulin ConcentidtioIl and hypochaldmic NP'Y mRNA levels (r=-0.39, p < 0 . 0 1 ) . These findings exclude the possibility that hyperphagia is the cause of the rise in hypothalamic NPY expression and argue that insulin may be an important inhibitory regulator of NPY expression in the ARC in lactation. this, lactating animals ale usually

Relationships between hypothalamic neuropeptide Y and food intake in the lactating rat.

NPYergic neurons in the hypothalamic arcuate nucleus (ARC) that project to the paraventricular nucleus (PVN) are postulated to regulate food intake and energy balance. This projection is overactive in lactation and is thought to drive hyperphagia in this condition. We have explored further the relationship between hypothalamic NPY and food intake in lactation and tested the hypothesis that hypoinsulinemia is the stimulus to NPY neuronal activity. Compared with nonlactating controls (n = 10), freely fed lactating rats (n = 9) showed significantly increased (p < 0.05) NPY levels in the ARC and medial preoptic area (MPO), but there was no significant increase in whole hypothalamic NPY mRNA levels. Lactating rats (n = 8) that were restricted to control rats' food intake for 3 days showed generally higher hypothalamic NPY levels, with significantly higher concentrations than controls (p < 0.05) in the ARC, MPO, PVN, and lateral hypothalamic area (LHA); NPY mRNA levels were also significantly increased (p < 0.05). Across all three experimental groups, there was a significant inverse correlation between plasma insulin concentration and hypothalamic NPY mRNA levels (r = -0.39, p < 0.01). We conclude that the ARC-PVN projection is overactive in lactation and that this is not a consequence of hyperphagia. Hypoinsulinemia may stimulate these neurons, as it is thought to do in other conditions of energy deficit.

Monosodium glutamate increases NGF and NPY concentrations in rat hypothalamus and pituitary

The effects of MSG treatment on NGF and NPY levels were analysed in the hypothalamus, pituitary, adrenal, thyroid and testis of adult rats. Daily i.v. injections of MSG (1 g kg-1 for 1 week) induced an increase of NGF in the hypothalamus (control (C) = 378 +/- 54; saline (S) = 369 +/- 36; MSG = 479 +/- 35 pg g-1 tissue; p < 0.001) and pituitary (C = 310 +/- 34; S = 376 +/- 114; MSG = 576 +/- 98 pg g-1 tissue; p < 0.01). Hypothalamic and pituitary NPY concentrations were also altered in the MSG-treated rats. Compared with saline-treated rats, the NPY concentration increased by 43% in the hypothalamus and 37.5% in the pituitary of MSG-treated rats. No significant changes in NGF and NPY content were found in the adrenal or thyroid of treated animals. These results suggest that hypothalamic and pituitary NGF and NPY levels may be involved in the control of neuroendocrine functions that are affected by MSG treatment.

Expression of pituitary adenylate cyclase-activating polypeptide in dorsal root ganglia following axotomy: time course and coexistence

Pituitary adenylate cyclase-activating polypeptide (PACAP) has recently been demonstrated in sensory neurons. In the present study on rat 17.5% of all neurons, mainly of small size, contained PACAP in normal dorsal root ganglia (DRGs). Transection of the sciatic nerve induced a rapid and strong upregulation in PACAP peptide and mRNA levels which could be seen already after 15 h. After 3 days more than 51.5% of neurons of different sizes expressed PACAP. However, the intensity of PACAP-LI in the DRG neurons declined after 10 days. Thirty days after axotomy, 56.7% of the DRG neurons still expressed PACAP, but with a low intensity, in fact even lower than in normal controls. No VIP- or NPY-positive neurons were observed in normal or axotomized DRGs at 15 h. However a distinct increase in VIP and NPY levels were seen 3 days after the lesion, and their levels were considerably higher after 30 days. PACAP was often present in neurons expressing VIP, NPY and or galanin. Thus, 3 days after injury, PACAP was present in 84.4%, 95.7%, and 76.8% of the VIP-, NPY-, and galanin-positive neurons, respectively. PACAP was also found in nerve fibers in control sciatic nerves. After nerve ligation, accumulation of PACAP was seen mainly proximal to the injury but also distally, suggesting both anterograde and retrograde transport of the peptide. Also a moderate increase (about 20%) in PACAP levels was found in the superficial spinal dorsal horn 3 days after nerve transection. Taken together, our results suggest that PACAP is involved in the response to nerve injury. The very high levels of expression in different populations of DRG neurons after axotomy, and its different time course as compared to galanin, NPY and VIP indicate that it may play a complementary and/or different role than these peptides in the adaptation to nerve injury, especially in its early phase.

Neuropeptide Y. A novel sympathetic stress hormone and more.

Several lines of evidence suggest that NPY is a neurotransmitter and neurohormone intricately involved in stress responses of the body, and as such should be considered a "stress molecule." Thus, circulating plasma NPY levels are increased by stress particularly if it is severe or prolonged. Stress stimulates the release of NPY from the sympathetic nerves and the adrenal medulla (in some species also from platelets), and in addition, modulates NPY inactivation. Stress-induced plasma NPY levels may reach the concentrations that are vasoconstrictive per se in addition to potentiating the actions of catecholamines. Reciprocally, elevated circulating levels of catecholamines during stress appear to induce hypersensitivity of blood vessels to NPY. Consequently, the peptide may be responsible for stress-induced regional vasoconstriction (splanchnic, coronary, and cerebral) but also may exert other actions that may be a part of the stress response: facilitate platelet aggregation, leukocyte adhesion, and macrophage activation. NPY release and actions appear to be up-regulated by testosterone and down-regulated by estrogens; therefore, NPY may be of particular importance to stress-induced cardiovascular events in men. In addition to acute vasoconstrictive effects, NPY exerts chronic actions and stimulates vascular smooth muscle proliferation and vascular hypertrophy, and hence, may be a link between stress and potential chronic changes in blood vessels.

Increased neuropeptide Y secretion in the hypothalamic paraventricular nucleus of obese (fa/fa) Zucker rats

NPY is synthesized in the hypothalamic arcuate nucleus (ARC), and NPY injected into the paraventricular nucleus (PVN), the main site of NPY release, induces hyperphagia and reduces energy expenditure. Hypothalamic NPY and mRNA and NPY levels are increased in fatty Zucker rats, consistent with increased NPY release. This could explain the hyperphagia and reduced energy expenditure, which lead to obesity in the fatty Zucker rat. We have therefore compared NPY secretion in the PVN of conscious fatty and lean Zucker rats using push-pull sampling. The NPY secretory profile was consistently higher in fatty Zucker rats than in lean rats throughout the 3-h study period ( P < 0.01), and mean NPY secretion over the whole 3 h was increased 2-fold in the fatty rats ( P < 0.001). We conclude that fatty Zucker rats have increased NPY release in the PVN. This observation further supports the hypothesis that increased activity of the NPYergic ARC-PVN pathway may contribute to obesity in the fatty Zucker syndrome.

Evidence that progesterone modulates anterior pituitary neuropeptide Y levels during the progesterone-induced gonadotropin surge in the estrogen-primed intact immature female rat.

In a previous study we reported that in vivo estrogen-priming alone, without subsequent progesterone-treatment, was sufficient to maximize NPY potentiation of gonadotropin hormone-releasing hormone responsiveness exhibited in vitro by the rat anterior pituitary. This observation suggests that the necessity, as reported by others, for both estrogen-priming and progesterone-treatment to maximize NPY potentiation of GnRH responsiveness in vivo may be due to progesterone acting primarily at the hypothalamus. Consequently, the current study was performed to determine whether progesterone facilitates gonadotropin secretion in vivo by acting to stimulate hypothalamic synthesis of NPY and the subsequent elevation of anterior pituitary tissue levels of NPY. Intact immature female rats were injected with estradiol at 1700 h on days 27 and 28. On day 29 at 0900 h, the animals received an injection of progesterone (2 mg/kg) or vehicle and were subsequently sacrificed at 1200, 1330 and 1500 h. Rats which received only estradiol injections were used as controls. Surge levels of serum LH and FSH were observed at 1330 and 1500 h. Hypothalamic levels of NPY mRNA at 1200 h on day 29 were higher (P < 0.01) in estradiol-primed rats which received progesterone; there was no accompanying statistically significant change in hypothalamic NPY content. NPY content in the anterior pituitary was significantly increased (P < 0.01) at 1200 h on day 29 in estradiol-primed rats which received progesterone; there was no accompanying significant change in anterior pituitary NPY mRNA levels. Hypothalamic GnRH mRNA content was significantly increased (P < 0.01) at 1330 h on day 29 concomitant with the peak of the gonadotropin surge in the estradiol-primed, progesterone-treated rat. The data indicate that progesterone modulates hypothalamic NPY mRNA and anterior pituitary NPY levels as well as GnRH mRNA levels and that modulation of NPY levels in the hypothalamic-pituitary axis occurs prior to modulation of GnRH gene expression. These studies support the hypothesis that in the estrogen-primed rat, progesterone facilitates the induction of the gonadotropin surge by maintaining hypothalamic synthesis of NPY as well as by modulating anterior pituitary NPY tissue levels.

Neuropeptide Y: distribution of immunoreactivity and quantitative analysis in diencephalic structures and cerebral cortex of dwarf hamsters under different photoperiods.

The distribution of neuropeptide Y-like immunoreactivity (NPY-LI) was investigated by immunohistochemistry and radioimmunoassay (RIA) in the brain of the Djungarian hamster (Phodopus sungorus) held under either long or short photoperiods. In the diencephalic and telencephalic structures studied, distinct patterns of NPY-LI were basically consistent in male and female animals of both groups. NPY levels detected by RIA from tissue samples taken at six time points throughout the 24-hour cycle were in the range of 15-60 pmol/mg protein in the diencephalon or below 5 pmol/mg protein in cerebral cortex. In the diencephalon, immunoreactive structures were seen in the preoptic, peri- and paraventricular, supraoptic, anterior, lateral, dorso- and ventromedial hypothalamic nuclei and in the median eminence. The suprachiasmatic nuclei exhibited a dense innervation by NPY-LI terminals mainly in its ventrolateral subdivision. NPY levels in the suprachiasmatic nucleus were nocturnally augmented under long-day, but not under short-day conditions. The quantification of NPY in the paraventricular nucleus revealed a decrease at night in long-day animals and a small nocturnal augmentation in short-day hamsters. In the pineal gland and habenular nuclei, varicose fibers were observed which appeared mainly perivascular in location (pineal) or formed a dense plexus (habenular nuclei). Pineal NPY contents fell during the night in long-day animals and were relatively constant in short-day hamsters. NPY-LI structures were also observed in the metathalamic intergeniculate leaflet and in a variety of telencephalic structures including the cerebral cortex, caudate nucleus putamen, lateral septal nucleus, bed nucleus of the stria terminalis and amygdala.

Photic information coded by vasoactive intestinal polypeptide and neuropeptide Y

We investigated photic response of the concentration of vasoactive intestinal polypeptide (VIP) and neuropeptide Y in the rat suprachiasmatic nucleus (SCN) by enzyme immunoassay (EIA). The content of VIP in the SCN did not show circadian rhythms in constant darkness (DD). Under light-dark (LD) condition, VIP contents decreased over the course of the light period and then recovered during the dark period. When the light was continuously delivered to rats, VIP levels were monotonically decreased and did not return to the basal level. Accordingly, VIP in the SCN may code photic information on duration. On the other hand, there is a daily bimodal pattern in NPY content in the SCN under light-dark conditions. When rats were exposed to continuous light, the NPY level steadily increased and reached a peak in 2 h before returning to a basal level. The amount of increase did not depend on duration of light exposure. Thus, NPY in the SCN may code visual information on transitions, which is different from that conveyed by VIP. These results indicate that the two peptides act on a different stage of photic processing and may mediate distinct photic information to the circadian pacemaker.

Hypothalamic concentration and release of neuropeptide Y into microdialysates is reduced in anorectic tumor-bearing rats

Hypothalamic concentration of neuropeptide Y was decreased significantly in anorectic tumor-bearing rats, while NPY level was increased significantly in matched carcass weight control rats as compared with freely-feeding controls. In vivo microdialysis of the perifornical hypothalamic area of tumor-bearing rats prior to the development of anorexia revealed no alteration in NPY in dialysates. Following the development of anorexia, however, tumor-bearing rats exhibited significant reduction in NPY concentration in dialysates as compared with either matched carcass weight or freely-feeding control group. These results suggest that hypothalamic NPY concentration and release are decreased selectively in anorectic tumor-bearing rats. Since NPY also elicits less feeding in tumor-bearing rats, dysfunction of hypothalamic NPY feeding mechanisms may be of primary importance in cancer anorexia.

Microdialysis in the estimation of interstitial myocardial neuropeptide Y release

The purpose of this study was to investigate the feasibility of cardiac microdialysis for the in vivo estimation of cardiac interstitial peptide concentrations, and, to determine the changes in neuropeptide Y release in myocardial tissue during experimental brain death in pigs. Using a specifically designed concentric flexible probe, perfused with Ringer solution containing 0.5% of bovine serum albumin at a flow rate of 2 μl/min, allowed us to obtain a 23 ± 2% relative recovery rate in vitro. Based on these in vitro recovery data, a regional study of the kinetics of interstitial NPY levels following brain death was obtained by monitoring the changes in NPY dialysate levels recorded from dialysis probes implanted into the right and left ventricular walls of the beating heart in vivo. Basal dialysate NPY levels determined by radioimmunoassay were of 95.2 ± 7.0 and 93.2 ± 9.1 pmol/l in left and right ventricle, respectively. Brain death was followed by a sustained 2 h increase in NPY dialysate levels in both ventricles (peak levels: 173.2 ± 30.9 pmol/l in left ventricle, and 149.7 ± 23.9 pmol/l in right ventricle), which then returned to control levels. We conclude that cardiac microdialysis is a simple and promising new tool for evaluating the role of peptides in cardiovascular regulation.

Chronic intracerebroventricular neuropeptide-Y administration to normal rats mimics hormonal and metabolic changes of obesity.

Chronic intracerebroventricular (icv) administration of neuropeptide-Y (NPY; 10 micrograms/day) was performed in normal female rats to investigate its hormonal and metabolic consequences. Intracerebroventricular NPY produced hyperphagia, increased basal insulinemia, as well as liver and adipose tissue lipogenic activity. It also increased basal morning corticosteronemia. When NPY-induced hyperphagia was prevented by pair-feeding, the icv NPY treatment resulted in the same increases in basal insulinemia and corticosteronemia, and liver and white adipose tissue lipogenesis was still higher than that in respective controls. Under the ad libitum and pair-feeding conditions, icv NPY stimulated glucose uptake as well as total lipoprotein lipase activity in white adipose tissue; it resulted in an increase total activity of hepatic and white adipose tissue acetyl coenzyme-A-carboxylase. As all hormonal and metabolic changes elicited by icv NPY remained present (at the same or to a lesser extent depending upon the parameter considered) when hyperphagia was prevented by pair-feeding, it was, thus, shown that icv NPY per se induces peripheral hormonal and metabolic alterations via efferent routes, which remain to be determined. The effects of icv NPY reported in this study are similar to the defects observed in the early phase of genetic obesity in rodents, the hypothalamus of which has increased NPY levels. NPY could, thus, be of relevance in the occurrence of genetically induced obesity.

Neuropeptide Y Concentrations in Cerebrospinal Fluid are Unchanged in Obesity

Neuropeptide Y (NPy) is a potent centrally acting appetite-stimulating peptide implicated in the regulation of energy balance. It induces hyperphagia and obesity when injected into the rat hypothalamus. Hypothalamic NPY and NPY mRNA levels are increased in spontaneously obese rats, suggesting that it may be involved in causing obesity in rodents. It is not known whether NPY has a role in the pathogenesis of obesity in man. NPY is found in human cerebrospinal fluid (CSF) and we have therefore compared CSF NPY levels in normal obese and non-obese individuals to determine whether NPY concentrations might be increased in obesity. We studied 25 clinically normal subjects (age 67 ± 5 years. male 11. female 14) undergoing spinal anaesthesia. None had any significant illness. Samples of 1 ml were freeze-dried and reconstituted to 100 ul and 35ul aliquots were assayed for NPY using an in-house RIA. CSF NPY levels were not correlated with body mass index (BMI) (r=O.088. p=O.673) and there were no differences in NPY concentrations between groups of subjects stratified for BMI: BMI 25 (n=ll), 702 ± 55 fmol/ml (differences between all groups. p>0l). CSF NPY levels are therefore not increased in human obesity. NPY is found in many brain regions outside the hypothalamic appetite-regulating nuclei, which could contribute to CSF levels. This negative observation does not therefore exclude a role of the peptide, acting specifically in the hypothalamus, in contributing to human obesity.

Acute hyperinsulinemia increases neuropeptide Y concentrations in the hypothalamic arcuate nucleus of fasted rats

Neuropeptide Y, a major hypothalamic peptide, stimulates feeding, insulin secretion and weight gain when injected intrahypothalamically. Hypothalamic NPY may be regulated by insulin availability at hypothalamic level, as its activity is apparently inhibited by intrahypothalamic insulin administration and is stimulated under insulin-deficient conditions. To determine the effects of acute physiological hyperinsulinemia, we measured regional hypothalamic NPY levels in rats during a hyperinsulinemic, euglycemic clamp. Seven male Wistar rats with implanted jugular cannulae, fasted for 24 h, were infused with insulin at 100 mU/h together with variable-rate glucose to maintain euglycemia (3.9±0.1 mmmol/l), for 150 min. Controls were infused for the same period with polygeline vehicle alone (n=8), and had blood glucose concentrations of 4.0±0.5 mmol/l. Insulin levels were 80.2±3.9 mU/l in insulin-infused rats and 15.2±1.4 mU/l in polygeline-treated controls (p<0.001). NPY levels, measured by radioimmunoassay, were significantly higher in the arcuate nucleus/median eminence (ARC/ME) of hyperinsulinemic rats than in controls (4.8±1.2 vs 2.5±0.6 fmol/ μg protein; p< 0.001), but were comparable with controls in 7 other hypothalamic regions. Acute physiological hyperinsulinemia therefore increases NPY levels selectively in the ARC/ME. Insulin could cause NPY accumulation in the ARC by blocking its transport to NPY-sensitive areas. This would be consistent with the suggestions that insulin inhibits hypothalamic NPY activity and also acts as a central satiety factor.

Characterization of phencyclidine-induced effects on neuropeptide Y systems in the rat caudate-putamen

Multiple administrations of the psychotominetic drug, phencyclidine-HC1 (PCP), decreased striatal neuropeptide Y-like immunoreactivity (NPY-LI) levels in a dose-dependent manner. Single or multiple PCP administrations decreased striatal NPY levels after 10–12 h; levels returned to control 24 h after a single dose or 58 h after multiple doses. In contrast, no significant changes were seen in nigral NPY levels with either acute or multiple-dose PCP treatments. The role of monoamine, σ or opioid receptors in PCP-induced striatal NPY changes was evaluated. When administered alone, the α 1-adrenergic antagonist, prazosin, the σ antagonist, BMY 14802, and the dopamine D 2 antagonist, sulpiride decreased striatal NPY levels; however, only prazosin and the dopamine D 1 antagonist, SCH 23390, significantly attenuated PCP-induced changes. Administration of the γ-aminobutyric acid transaminase (GABA-T) inhibitors, amino-oxyacetic acid (AOAA) or γ-vinyl-GABA (GVG, vigabatrin, MDL 71,754) alone had no effect on striatal NPY-LI levels while administration of these indirect GABA agonists prior to or concurrently with PCP treatment completely blocked PCP-induced changes in striatal NPY-LI levels. The effect of the non-competitive N- methyl- d- aspartate (NMDA) receptor antagonist, MK-801, on striatal NPY-LI content resembled that of PCP and was also blocked by the two indirect GABA agonists. These data suggest that NPY systems are modulated by glutamatergic activity (specifically by the NMDA receptor) and that the interaction between these two transmitter systems is mediated by GABAergic mechanisms.