Neuropeptide Y Immunoreactivity Research Articles

Ontogeny of the endocrine pancreatic cells of the gilthead sea bream, Sparus aurata (Teleost)

The development of the gilthead sea bream, Sparus aurata, endocrine pancreas was studied from hatching to 114 days, using immunocytochemical techniques. Bonito insulin (INS)-, synthetic somatostatin-14 (SS-14)-, salmon somatostatin-25 (SS-25)-, mammalian somatostatin-28 (1–12) (SS-28)-, porcine glucagon (GLU)-, glucagon-like peptide-1 (1–19) (GLP-1)-, synthetic porcine peptide tyrosine tyrosine (PYY)-, and neuropeptide Y (NPY)-like immunoreactivities were demonstrated. The different types of endocrine cells appear at distinct stages of development and differ in their arrangement. The coexistence of INS and SS-25 immunoreactivities was demonstrated in the cells of one strand or primordial cord and a primordial islet that appeared close to the dorsal epithelium of the anterior region of the undifferentiated gut or next to the gut at hatching and one day after hatching, respectively. INS- and SS-25- immunoreactive (ir) cells were located in the core and at the periphery of the single islet found in 2-day-old larvae, while SS-28-ir cells were found in the single islet in 4- to 11-day-old larvae. GLU/GLP-1-ir cells were located next to the outer SS-25-ir cells in the single islet of 12- and 16-day-old larvae. SS-14/SS-25- and SS-14/SS-28-ir cells were detected in the outer region and in the inner area of the single islet, respectively, in 17- to 23-day-old larvae. One big islet and several small islets and isolated or clustered cells next to the pancreatic duct were present in 24- and 25-day-old larvae. The islets were similar in cell composition to the single islet seen in the previous stage, while the isolated and grouped cells showed the coexistence of INS and diverse SSs immunoreactivities. Nerve fibers showing PYY immunoreactivity were identified in the islets from 17 days onwards. In 30- to 44-day-old larvae, GLU and NPY immunoreactivities coexisted in a few cells at the periphery of some small islets. PYY-ir cells were first detected at day 51. One big islet, several intermediate islets and numerous small islets were present from 51-day-old-larvae to juveniles. GLU was colocalized with PYY and NPY in a few cells in a small peripheral area in the big islet and a few intermediate islets. The outer region of small islets and other intermediate islets showed the complete coexistence of GLU, PYY, and NPY.

Sympathetic Neuronal‐Megakaryocyte Axis: a key to stress‐induced amplification of atherosclerotic‐like lesions

While usually described together, the true link between stress and the onset of atherosclerosis has yet to be elucidated. Neuropeptide Y (NPY), a sympathetic neurotransmitter and vascular growth factor, exhibits a human gene polymorphism associated with accelerated atherosclerosis, and in mice, induces atherosclerotic-like lesions after angioplasty. We have found that NPY is a mitogen for smooth muscle cells in vitro. Further, platelet NPY immunoreactivity levels increase after stress by approximately 12%. These findings suggest megakaryocytes influence the increase in NPY mRNA after stress, as detected by quantitative RT-PCR. Sympathetic activation and NPY release was induced by cold stress of 1hr/day/14days in 4°C water, and when combined with angioplasty, resulted in increased neointimal formation and vascular occlusive lesions. Therefore, we hypothesize that stress promotes vascular occlusion after angioplasty by activating a novel neuronal-megakaryocyte axis, leading to increased platelet NPY levels, which is pro-atherosclerotic. Time trials of 1, 3, 7, and 14 days resulted in a 2–4 fold increase of neointimal formation over time, as compared to NPY-KO studies. Platelet transfer experiments from wild-type to NPY KO mice (and vice-versa) validated the role of platelet NPY in neoinitmal formation after angioplasty. We conclude that stress up-regulates NPY expression in megakaryocytes, likely via a sympathetic link, as shown by positive immunohistochemical co-localization staining for synapsin, CD41 and tyrosine hydroxylase in bone marrow, leading to increased platelet NPY and neointimal formation. Thus, platelet NPY may serve as an important factor linking stress to atherosclerosis after angioplasty.

Differential responses of the growth hormone axis in two rat models of streptozotocin-induced insulinopenic diabetes

The impact of streptozotocin (STZ)-induced, insulinopenic diabetes on the GH axis of rats and mice differs from study to study, where this variation may be related to the induction scheme, severity of the diabetes and/or the genetic background of the animal model used. In order to begin differentiate between these possibilities, we compared the effects of two different STZ induction schemes on the GH axis of male Sprague-Dawley rats: (1) a single high-dose injection of STZ (HI STZ, 80 mg/kg, i.p.), which results in rapid chemical destruction of the pancreatic beta-cells, and (2) multiple low-dose injections of STZ (LO STZ, 20 mg/kg for 5 consecutive days, i.p.), which results in a gradual, autoimmune destruction of beta-cells. STZ-treated animals were killed after 3 weeks of hyperglycemia (>400 mg/dl), and in both paradigms circulating insulin levels were reduced to <40% of vehicle-treated controls. HI STZ-treated rats lost weight, while body weights of LO STZ-treated animals gradually increased over time, similar to vehicle-treated controls. As previously reported, HI STZ resulted in a decrease in circulating GH and IGF-I levels which was associated with a rise in hypothalamic neuropeptide Y (NPY) mRNA (355% of vehicle-treated controls) and a fall in GH-releasing hormone (GHRH) mRNA (45% of vehicle-treated controls) levels. Changes in hypothalamic neuropeptide expression were reflected by an increase in immunoreactive NPY within the arcuate and paraventricular nuclei and a decrease in GHRH immunoreactivity in the arcuate nucleus, as assessed by immunohistochemistry. Consistent with the decline in circulating GH and hypothalamic GHRH, pituitary GH mRNA levels of HI STZ-treated rats were 58% of controls. However, pituitary receptor mRNA levels for GHRH and ghrelin increased and those for somatostatin (sst2, sst3 and sst5) decreased following HI STZ treatment. The impact of LO STZ treatment on the GH axis differed from that observed following HI STZ treatment, despite comparable changes in circulating glucose and insulin. Specifically, LO STZ treatment did suppress circulating IGF-I levels to the same extent as HI STZ treatment; however, the impact on hypothalamic NPY mRNA levels was less dramatic (158% of vehicle-treated controls) where NPY immunoreactivity was increased only within the paraventricular nucleus. Also, there were no changes in circulating GH, hypothalamic GHRH or pituitary receptor expression following LO STZ treatment, with the exception that pituitary sst3 mRNA levels were suppressed compared with vehicle-treated controls. Taken together these results clearly demonstrate that insulinopenia, hyperglycemia and reduced circulating IGF-I levels are not the primary mediators of hypothalamic and pituitary changes in the GH axis of rats following HI STZ treatment. Changes in the GH axis of HI STZ-treated rats were accompanied by weight loss, and these changes are strikingly similar to those observed in the fasted rat, which suggests that factors associated with the catabolic state are critical in modifying the GH axis following STZ-induced diabetes.

Effect of maternal thyroxine treatment on the offspring's brain development with fetal alcohol effects in the rats

Purpose:This study aimed to investigate whether exogenous thyroxine(T4) treatment to alcohol-fed dams would ameliorate the detrimental effects of alcohol on the postnatal development of neuropeptide-Y(NPY)-containing neurons of the cerebral cortex and hippocampus of the offspring. Methods:Time-pregnant rats were divided into three groups. An alcohol-fed group A received 35 calories of liquid alcohol diet daily from gestation day 6; control group B was fed a liquid diet in which dextrin replaced alcohol isocalorically; and alcohol＋T4 group C received 35 calories of liquid alcohol diet and exogenous thyroxine subcutaneously. The features of the growth and maturation of rat brain tissue were observed at 0, 7, 14, 21 and 28 postnatal days via immunohistochemistry. Results:Group C showed prominent NPY immunoreactivity in the cerebral cortex compared to group A and B at P7. In group C, NPY-containing neurons were widely distributed in the all layers of cerebral cortex after P14. Also, numerical decreases of NPY-containing neuron were not found according to increasing age in group C. A decrease of NPY-containing neurons, however, was clearly observed in group A compared to group C at P28. In the hippocampus, similar patterns appeared in groups B and C after P7. Especially, in groups B and C, NPY-containing fibers formed plexus in the cerebral cortex and hippocampus at P14. Conclusion:These results suggest that the increase of NPY synthesis caused by maternal administration of exogenous thyroxine may convalesce fetal alcohol effects, one of the effects of the dysthyroid state following maternal alcohol abuse.

Differential effects of vigabatrin and zonisamide on the neuropeptide Y system in the hippocampus of seizure prone gerbil

Changed neuropeptide Y (NPY) system in the hippocampus has been reported in various experimental epileptic models. However, there have been little data concerning the alteration in the NPY system in the epileptic hippocampus following treatment of anti-epileptic drugs (AEDs). In the present study, therefore, we performed analyses of effects of vigabatrin (VGB) and zonisamide (ZNS) treatment on the NPY system in the hippocampus of the seizure sensitive (SS) gerbils. In SS gerbil, NPY immunoreactivity in the hippocampus was lower than that in seizure resistant gerbil. Following VGB treatment, the number of NPY immunoreactive neurons and NPY mRNA expression were increased in the hilus and the hippocampus proper. In contrast, ZNS treatment markedly elevated only the density of NPY immunoreactive fibers in the dentate gyrus, not in the hippocampus proper, as compared with saline-treated animals. These patterns were observed in the dose-dependent manners. These findings suggest that AEDs treatments may distinctly affect the NPY system in the SS gerbil hippocampus.

Influence of cold stress on neuropeptide Y and sympathetic neurotransmission

Chronic cold stress of rats (4 °C; 1–3 weeks) induced a marked increase in gene expression (adrenal medulla; superior cervical ganglia), tissue content (mesenteric arterial bed) and nerve stimulation-induced overflow of NPY-immunoreactivity (NPYir) from the perfused mesenteric arterial bed. In contrast increased NPY neurotransmission was offset by an apparent decrease in the evoked overflow of norepinephrine (NE) due to a presumed deactivation of NE by nitric oxide (NO), despite increased sympathetic nerve activity. The net effect of these offsetting system was no change in basal or the evoked increase in perfusion pressure (sympathetic tone). It is concluded that differences in NPY and NE transmission act as an important compensatory mechanism preventing dramatic changes in arterial pressure when sympathetic nerve activity is high during cold stress.

Neuropeptide Y in the forebrain of the adult male cichlid fish Oreochromis mossambicus: Distribution, effects of castration and testosterone replacement

We studied the organization of the neuropeptide Y (NPY)-immunoreactive system in the forebrain of adult male cichlid fish Oreochromis mossambicus and its response to castration and testosterone replacement by using morphometric methods. Immunoreactivity for NPY was widely distributed in the forebrain, and the pattern generally resembled that in other teleosts. Whereas immunoreactivity was conspicuous in the ganglia of nervus terminalis (NT; or nucleus olfactoretinalis), a weak reaction was detected in some granule cells in the olfactory bulb and in the cells of area ventralis telencephali pars lateralis (Vl). Moderately to intensely immunoreactive cells were distinctly seen in the nucleus entopeduncularis (NE), nucleus preopticus (NPO), nucleus lateralis tuberis (NLT), paraventricular organ (PVO), and midbrain tegmentum (MT). NPY fibers were widely distributed in the forebrain. Castration for 10/15 days resulted in a drastic loss of immunoreactivity in the cells of NE (P<0.001) and a significant decrease (P<0.01) in their cell nuclear size. However, cell nuclei of the NT neurons showed a significant increase in size. A highly significant reduction in the NPY-immunoreactive fiber density (P<0.001) was observed in several areas of the forebrain. Although testosterone replacement reversed these changes, fibers in some areas showed supranormal responses. Immunoreactive cells in Vl, NPO, NLT, PVO, and MT and fiber density in some other areas did not respond to castration. We suggest that the NPY-immunoreactive elements that respond to castration and testosterone replacement may serve as the substrate for processing the positive feedback action of the steroid hormone.

Neuropeptide Y expression, localization and cellular transducing effects in HUVEC

NPY (neuropeptide Y) may have an effect on the properties of vascular endothelial cells such as pro-angiogenic effects and potentiation of noradrenaline-induced vasoconstriction. In HUVEC (human umbilical-vein endothelial cells), immunoreactive neuropeptide Y has been detected, but NPY synthesis, storage and secretion have not been studied. The aim of the present study was to establish NPY expression, storage and cellular transducing effects in HUVEC. HUVEC contain 0.19 fmol of NPY/microg of protein and 0.46 fmol of pro-NPY/microg of protein, as measured by ELISA. RT (reverse transcriptase)-PCR confirmed the expression of NPY in HUVEC. Immunofluorescence revealed the presence of NPY in small punctate structures, with a fluorescence pattern different from that observed for von Willebrand factor, indicating distinct storage compartments. Double labelling for NPY and Rab3A demonstrated similar granular patterns, with at least partial co-localization. Electron microscopy showed NPY immunoreactivity in vesicle-like cytoplasmic structures, of a fine fibrillar texture, as well as in mitochondria and in the nucleus. A similar general distribution pattern was also obtained for Rab3A. Y1 and Y2 receptors were expressed in HUVEC as assessed by RT-PCR, and they were functional since NPY induced a 42 nM intracellular calcium increase within 100 s, representing 22% of the histamine-induced response. In contrast with histamine, NPY did not induce acute von Willebrand factor secretion. HUVEC produce, store and respond to NPY, suggesting an autocrine regulatory role for NPY in the endothelium.

Comparison of Basal Neuropeptide Y and Corticotropin Releasing Factor Levels Between the High Ethanol Drinking C57BL/6J and Low Ethanol Drinking DBA/2J Inbred Mouse Strains

Recent genetic and pharmacological evidence indicates that low neuropeptide Y (NPY) levels in brain regions involved with neurobiological responses to ethanol promote increased ethanol consumption. Because of their opposing actions, it has been suggested that NPY and corticotropin releasing factor (CRF) exert a reciprocal regulation on drug self-administration. It has been widely reported that inbred C57BL/6 mice consume significantly higher amounts of ethanol than do DBA/2 mice. Therefore, we used immunohistochemical techniques to determine if basal NPY and/or CRF levels differed in predicted directions between C57BL/6J and DBA/2J mice. Ethanol-naive C57BL/6J and DBA/2J mice were deeply anesthetized with sodium pentobarbital (100 mg/kg) and perfused transcardially with 0.1 mM of phosphate-buffered saline followed by 4% paraformaldehyde in buffered saline. Brains were collected and postfixed for 4 hr at 4 degrees C and then were cut into 35-microm sections. Tissues containing the nucleus accumbens (NAc), hypothalamus, and amygdala were processed for NPY or CRF immunoreactivity using immunofluorescent or DAB techniques. Immunoreactivity was quantified from digital images using Image J software. The C57BL/6J mice showed reduced NPY expression in the NAc shell, the basolateral amygdala, and the central nucleus of the amygdala when compared with DBA/2J mice. However, these strains did not differ in CRF expression in any of the brain regions analyzed. These data suggest that low NPY levels in the amygdala and/or the shell of the NAc, which are not compensated for by similar changes in CRF levels, may contribute to the high ethanol consumption characteristic of C57BL/6J mice.

Effects of omega-3 fatty acids on orexigenic and anorexigenic modulators at the onset of anorexia

In cancer anorexia, a decrease in food intake (FI) occurs concomitant with changes in orexigenic peptides such as neuropeptide Y (NPY) and anorexigenic peptides such as α-melanocyte-stimulating hormone (α-MSH) and anorexigenic neurotransmitter serotonin. ω-3 Fatty acid (ω-3FA) inhibits cytokine synthesis, and delays tumor appearance, tumor growth, and onset of anorexia in tumor-bearing rats. We hypothesize that, in cancer anorexia, ω-3FA is associated with quantitative reversal of hypothalamic NPY, α-MSH, and serotonin receptor (5-HT 1B-receptor) enhancing FI. Fischer rats were divided into: MCA tumor bearing fed chow (TB-Chow) or ω-3FA diet (TB-ω-3FA) and controls: non-tumor bearing fed chow (NTB-Chow) or ω-3FA diet (NTB-ω-3FA). Rats were euthanized at anorexia and brains were removed for hypothalamic immunohistochemical study, using NPY, α-MSH, and 5-HT 1B-receptor-specific antibodies and slides assessed by image analysis. Immunostaining specificity was controlled by omission of primary or secondary antibodies and pre-absorption test. At anorexia, FI decreased ( P < 0.05) in TB-Chow but did not change in TB-ω-3FA rats. In TB-ω-3FA vs. TB-Chow, NPY immunoreactivity increased 38% in arcuate nucleus (ARC; P < 0.05), and 50% in magnocellular paraventricular nucleus (mPVN; P < 0.05). α-MSH decreased 64% in ARC and 29% in mPVN ( P < 0.05). 5-HT 1B-receptor immunoreactivity decreased 13% only in supraoptic nucleus ( P < 0.05). No immunoreactivity was found in the control sections. ω-3FA modified hypothalamic peptides and 5-HT- 1B-receptor immunoreactivity at anorexia, concomitant with an increase in FI, were probably mediated by ω-3FA inhibition of tumor-induced cytokines.

Role of neuropeptide Y (NPY) in the regulation of reproduction: study based on catfish model

Significance of NPY in the regulation of GnRH-LH axis was evaluated. Considerable NPY immunoreactivity was seen in the components like olfactory system, basal telencephalon, preoptic and tuberal areas, and the pituitary gland that serve as neuroanatomical substrates for processing reproductive information. Close anatomical association as well as colocalizations of NPY and GnRH were seen in the olfactory receptor neurons, olfactory nerve fibers and their terminals in the glomeruli, ganglion cells of nervus terminalis, medial olfactory tracts, fibers in the ventral telencephalon and pituitary. In the pituitary, NPY fibers seem to innervate the GnRH as well as LH cells. Intracranial administration of NPY resulted in significant increase in the GnRH immunoreactivity in all the components of the olfactory system. In the pituitary, NPY augmented the population of GnRH fibers and LH cells. HPLC analysis showed that salmon GnRH content in the olfactory organ, bulb, preoptic area+telencephalon and pituitary was also significantly increased following NPY treatment. NPY may play a role in positive regulation of GnRH throughout the neuraxis and also up-regulate the LH cells in the pituitary.

Neuropeptide Y-immunoreactive (NPY-ir) structures in the brain of the gar Lepisosteus oculatus (Lepisosteiformes, Osteichthyes) with special regard to their anatomical relations to gonadotropin-releasing hormone (GnRH)-ir structures in the hypothalamus and the terminal nerve

The present paper describes neuropeptide Y-like-immunoreactive (NPY-ir) structures in the brain of the spotted gar, Lepisosteus oculatus, with special regard to their anatomical relations to gonadotropin-releasing hormone (GnRH)-ir structures in the hypothalamus and the terminal nerve (TN). NPY-ir cells were found in various locations including the TN, the medial zone of the area dorsalis telencephali, the ventral nucleus of the area ventralis telencephali, the habenula, the dorsal posterior nucleus, the periventricular nucleus of the hypothalamus, the posterior tubercle, the optic tectum, and the lateral part of the tegmentum. NPY-ir fibers were widely distributed throughout the brain except for the cerebellum. They were locally dense in the ventral telencephalon, in the periventricular gray matter of the thalamus and the hypothalamus, and in the ventromedial part of the brainstem, but sparse in the olfactory system. Light-microscopic double immunohistochemistry demonstrated distinct NPY-ir and GnRH-ir structures in the ventral hypothalamus: the NPY-ir system was associated mainly with the periventricular gray matter, whereas the GnRH-ir system was prominent in the external zone of the preoptico-tubero-infundibular area including the median eminence (ME). Here, NPY-ir varicose fibers occasionally abutted on GnRH-ir cells and varicosities or invested GnRH-ir cells, suggesting that NPY directly regulates the function of the hypothalamic GnRHergic neuron system. On the other hand, the TN cells and fibers in the olfactory system were doubly labeled by the antibodies against NPY and GnRH. Immuno-electron-microscopic data strongly suggested that some of the TN fibers projected to the ME.

CCK-8 prevents the development of kindling and regulates the GABA and NPY expression in the hippocampus of pentylenetetrazole (PTZ)-treated adult rats

Neuronal loss and irreversible brain damage often cause the worsening of symptoms and the decreased efficacy of pharmacological treatment occurring in epileptic patients and animal models of kindling. Recently we reported that the neurotransmitter/neuromodulatory peptide Cholecystokinin-8 (CCK-8) is able to induce the structural and functional neuronal recovery of chemical- and surgical-induced lesions when i.p. injected in rodents. The present study therefore, was aimed at verifying the hypothesis that treatment with a CCK-8 dose having a neuroprotective action might affect brain alterations and the development of kindling in adult rats receiving the convulsant agent pentylenetetrazole (PTZ). Compared to rats receiving Saline prior to PTZ, which manifested clonic–tonic seizures (Class 5 behavioural change scale) after three weeks of treatment, rats pre-treated with CCK-8 showed an improvement of behavioural score exhibiting myoclonus and occasionally tonic seizures (Class 3/4). This decreased susceptibility to develop convulsions was associated with the recovery of PTZ-induced reduction of ChAT levels in forebrain and GABA/GAD expression in the hippocampus. Furthermore, NPY immunoreactivity distribution and NPY mRNA levels were also increased in the hippocampus of rats receiving CCK-8 injection before each PTZ treatment. These data indicate that CCK-8 possesses the ability to prevent and/or suppress the convulsant effects of PTZ by stimulating the synthesis of neurotransmitters/peptides involved in the inhibition of hippocampal hyper-excitability. Our findings suggest that CCK-8 may have anticonvulsant and neuroprotective properties that merit further investigation.

Changes in Orexin-A and Neuropeptide Y Expression in the Hypothalamus of Obese and Lean Zucker Diabetic Fatty Rats

This study was carried out to investigate the changes of orexin-A (OXA) and neuropeptide Y (NPY) expression in the hypothalamus of the obese and lean Zucker Diabetic Fatty (ZDF) rats which have a missense mutation in the leptin receptor gene. The mean body weights (MBW) between the obese and lean ZDF rats were significantly different at 28 and 70 postnatal days. However, at 14 postnatal day, there was no significant difference in the MBW between the obese and lean ZDF rats in both male and female. The OXA immunoreactivities were not significantly different between the obese and lean ZDF rats in both sexes at 14, 28, and 70 postnatal days, respectively. The NPY immunoreactivity was higher in the obese than in the lean ZDF rats in both male and female at 28 and 70 postnatal days, whereas there was no significant difference between the obese and lean ZDF rats at 14 postnatal day. These results indicate that both OXA and NPY might halt their roles for food intake in the obese phenotype of the male and female ZDF rats in the preweaning period of 14 postnatal day, whereas NPY might play a main role in the obesity of these rats in the weaning period of 28 and 70 postnatal days.

The behavioral and neuroanatomical effects of IB 4-saporin treatment in rat models of nociceptive and neuropathic pain

One distinguishing feature of primary afferent neurons is their ability to bind the lectin IB 4. Previous work suggested that neurons in the inner part of lamina II (IIi), onto which IB 4-positive sensory neurons project, facilitate nociceptive transmission following tissue or nerve injury. Using an IB 4-saporin conjugate (IB 4-SAP), we examined the contribution of IB 4-positive neurons to nociceptive processing in rats with and without nerve injury. Intrasciatic injection of IB 4-SAP (5 μg/5 μl) significantly decreased IB 4-labeling and immunoreactive P 2X 3 in the spinal cord and delayed the behavioral and neuroanatomical consequences of L5 spinal nerve ligation (SNL) injury. In the absence of injury, thermal and mechanical nociceptive thresholds increased 2 weeks post-treatment only in IB 4-SAP-treated, but not control (saline or saporin only), rats. Acute NGF-induced hyperalgesia was also attenuated following IB 4-SAP treatment. In the SNL model, mechanical allodynia failed to develop 1 and 2 weeks post-injury, but was fully established by 4 weeks. Moreover, neuropeptide Y immunoreactivity (NPY-ir), which increases in the spinal cord after nerve injury, was unchanged in IB 4-SAP-treated animals whereas immunoreactive PKCγ decreased 2, but not 4, weeks post-injury. Quantitative RT-PCR revealed a reduction in P 2X 3 mRNA in L4 DRG of IB 4-SAP-treated animals, but no change in TrkA expression. Our results suggest that IB 4-positive neurons in L4 are required for the full expression of NGF-induced hyperalgesia and participate in the behavioral and anatomical consequences that follow injury to the L5 spinal nerve.

Distribution of neuropeptide Y-immunoreactive neurons in the human brainstem, cerebellum, and cortex during development.

1. Neuropeptide Y is found throughout the central nervous system where it appears to play a wide range of often poorly understood functions. In this study, the distribution of neuropeptide Y immunoreactive (NPY-ir) neurons in the brainstem, cerebellum, and cerebral cortex of human fetuses ranging in age from 11 gestational weeks to term was investigated by immunohistochemistry. 2. The NPY-ir cells were detected in the dorsal and ventral rostral midbrain and the interpeduncular nucleus by 21 weeks and 32 weeks of gestation, respectively. Although no positive cells were found in the pons, the NPY-ir fibers were detected there at 32 gestational weeks. 3. The vagal, hypoglossal, and olivary nuclei of the medulla oblongata contained immunoreactive cells by week 21 and the medullary reticular formation by week 25 of gestation. In most of these locations, both the number and size of neuropeptide Y positive cells were greater at birth and reached maximal values of 100-400 cells per 1 mm2 and 2-5 microm in diameter, respectively. 4. In the cerebellum, numerous NPY-ir horizontal and granule cells, as well as the cells within the dentate nucleus were observed as early as 21 weeks of gestation. 5. The NPY-ir cells were also detected in the developing cerebral cortex, with the earliest activity observed within the temporal cortex at 14 weeks of gestation. By week 21, positive cells appeared in the visual, frontal, sensory, and motor cortices. Most of these cells were bipolar or multipolar in morphology but their numbers at birth were relatively low. 6. Our results show a wide distribution of the NPY-ir cells in the developing human brain and offer supporting evidence for the important modulatory role of NPY in both the fetus and adult.

Immunohistochemical and genomic evidence for the involvement of hypothalamic neuropeptide Y (NPY) in phenylpropranolamine-mediated appetite suppression

Phenylpropanolamine (PPA) is an appetite suppressant. The mechanism for the anorectic effect of PPA has been attributed to its action on the site of hypothalamic paraventriculum. Neuropeptide Y (NPY) is an appetite stimulant that is widely distributed in the site of hypothalamus. It is not clear whether hypothalamic NPY is involved in the anorectic action of PPA. This study was aimed to investigate the mechanism underlying the involvement of NPY gene in the anorectic action of PPA. Results revealed that PPA treatment in rats could decrease both NPY content and mRNA level in the hypothalamus. In addition, the expression of NPY immunoreactivity following PPA treatment was decreased in areas of hypothalamic arcuate nucleus, paraventricular nucleus and periventricular area using immunohistochemical staining, suggesting an involvement of NPYergic pathway in the action of PPA anorexia. Our results provided immunohistochemical and genomic evidence to suggest that PPA might reduce feeding by altering NPY gene expression.

Immunohistochemical distribution of neuropeptide Y in the mesencephalon and rhombencephalon of carp, Cyprinus carpio L. (Cyprinidae: Teleostei)

The localization of neuropeptide Y (NPY)-immunoreactive elements was investigated in the mesencephalon and rhombencephalon of carp, Cyprinus carpio, by using antisera raised against porcine NPY and the immunoperoxidase technique. Concurrently, to identify the distribution of NPY-immunoreactivity, we developed an atlas of the studied areas based on Nissl-stained sections. The NPY-immunoreactive (NPY-ir) elements were located in many zones of the mesencephalon and rhombencephalon. In the mesencephalon, positive fibers were the most abundant elements while neurons were scarce. The rhombencephalon rostral part was characterized by a low to moderate fiber density, distributed in the ventro-medial and ventro-lateral region. Differently the caudal part of the rhombencephalon exhibited several NPY-ir elements. In particular, a high density of immunoreactivity was located in the gustatory area at the level of the nucleus (n.) originis nervi glossopharyngei, in the n. nervi vagi, and in the vagal lobe. The latter can be considered a valid neuroanatomical model for the study of gustatory signal processing in vertebrates. Our results regarding the primary gustatory centers give neuroanatomical support to the view that NPY may act as a neurotransmitter and/or a neuromodulator in a wide neural network for feeding behavior control.

Plasma neuropeptide Y immunoreactivity influences left ventricular mass in pheochromocytoma

Left ventricular hypertrophy (LVH) in patients with arterial hypertension is closely related to the levels of blood pressure (BP), catecholamines, angiotensin II and other mitogenic peptides. Pheochromocytoma (pheo) is a type of hypertension caused by excessive production of catecholamines. The aim of this study was to determinate if left ventricular hypertrophy in patients with pheochromocytoma is related to catecholamines and neuropeptide Y (NPY). Methods: 29 patients with pheochromocytoma (22 F, age 40±13 years), plasma concentration of neuropeptide Y immunoreactivity, noradrenaline (NA), and adrenaline (A) were determined. Twenty-four hour urine collection for determination of noradrenaline and adrenaline were performed. Every patient had echocardiographic examination and 24 h ambulatory blood pressure monitoring. Results: Left ventricular hypertrophy was diagnosed in 14 patients. No differences in systolic and diastolic blood pressure in patients with and without left ventricular hypertrophy were found. Plasma noradrenaline and adrenaline levels did not differ between both groups, while plasma neuropeptide Y immunoreactivity was higher in patients with left ventricular hypertrophy than in patients without left ventricular hypertrophy (18.46±13.26 vs. 9.3±5.9 fmol/ml ( p=0.02)). Left ventricular mass index (LVMI) correlated with plasma neuropeptide Y-immunoreactivity ( r=0.42 p=0.023), however, no relationship between left ventricular mass index and plasma or urine noradrenaline and adrenaline levels were found. Conclusion: Our results indicate that mitogenic effect of neuropeptide Y may play a role in pathogenesis of left ventricular hypertrophy in patients with pheochromocytoma.

Metabolic fuels, neuropeptide Y, and estrous behavior in Syrian hamsters

Of the various environmental factors influencing reproduction, food availability plays a particularly significant role, and an insufficient supply of oxidizable metabolic fuels inhibits reproduction in female mammals. When ovariectomized, steroid-primed hamsters are food deprived for 48 h, estrous behavior is suppressed. However, the specific neuroendocrine alterations that mediate the suppression of estrous behavior are unknown. Several conditions that inhibit female sexual behavior are thought to be associated with altered neuropeptide Y (NPY) activity in the brain. Intracerebroventricular (ICV) infusion of NPY inhibits estrous behavior in ovariectomized steroid-primed rats and hamsters. Furthermore, food-deprived rats have an increase in NPY mRNA in the arcuate nucleus (ARC) of the hypothalamus. Unlike rats, studies in Syrian hamsters have failed to detect any alterations in ARC NPY mRNA following food deprivation. Here we show that ARC NPY immunoreactivity and mRNA is increased in food-deprived hamsters but not in hamsters given other metabolic challenges that inhibit estrous behavior. These findings support the hypothesis that NPY contribute to, but not be critical for, the nutritional inhibition of sexual receptivity.