Effects of neuropeptide Y (NPY) on mechanical activity and neurotransmission in the heart, vas deferens and urinary bladder of the guinea-pig

The effects of preincubation for 10 min with synthetic porcine neuropeptide Y (NPY) on muscle tone and autonomic transmission in the guinea-pig right atrium, vas deferens, urinary bladder, portal vein and trachea were analysed in vitro. NPY induced a metoprolol-resistant, long-lasting, positive inotropic and chronotropic effect per se in the spontaneously beating right atrium. Furthermore, NPY caused a reversible inhibition of both the metoprolol and atropine-sensitive auricle responses to field stimulation (2 Hz or 4 Hz for 2 s) without affecting the response to exogenous noradrenaline (NA) or acetylcholine (ACh). NPY did not induce any contraction of the vas deferens, but inhibited both the rapid twitch response and the sustained tonic contraction induced by field stimulation. The NPY-induced inhibition of the tonic contraction was more long-lasting than that of the twitch response. The tonic contraction was blocked by phentolamine and the twitch response by alpha-, beta-methylene ATP tachyphylaxis. NPY did not inhibit the contractile effects of NA, ATP or alpha-, beta-methylene ATP. NPY also induced a reversible reduction of the non-cholinergic, non-adrenergic contractile response to field stimulation of the urinary bladder. In the portal vein, NPY (up to 5 X 10(-7) M) did not inhibit the spontaneous motility or the phentolamine-sensitive contractile responses to field stimulation and NA. The atropine-sensitive contraction of the trachea or the non-adrenergic, non-cholinergic relaxation induced by field stimulation were not significantly influenced by NPY in doses up to 5 X 10(-7) M.(ABSTRACT TRUNCATED AT 250 WORDS)     

New immunomodulatory role of neuropeptide Y (NPY) in Salmo salar leucocytes

Neuropeptide Y (NPY) plays different roles in mammals such as: regulate food intake, memory retention, cardiovascular functions, and anxiety. It has also been shown in the modulation of chemotaxis, T lymphocyte differentiation, and leukocyte migration. In fish, NPY expression and functions have been studied but its immunomodulatory role remains undescribed. This study confirmed the expression and synthesis of NPY in S. salar under inflammation, and validated a commercial antibody for NPY detection in teleost. Additionally, immunomodulatory effects of NPY were assayed in vitro and in vivo. Phagocytosis and superoxide anion production in leukocytes and SHK cells were induced under stimulation with a synthetic peptide. IL-8 mRNA was selectively and strongly induced in the spleen, head kidney, and isolated cells, after in vivo challenge with NPY. All together suggest that NPY is expressed in immune tissues and modulates the immune response in teleost fish.     

Effects of cervical sympathetic nerve stimulation and neuropeptide Y (NPY) on cranial blood flow in the cat

Effects of cervical sympathetic nerve stimulation (SNS) at 10 Hz and intravenous infusion of neuropeptide Y (NPY), 10 and 100 pmol x kg body wt-1 x min-1 for 5 min, on regional blood flow in the cat were investigated with radioactive microspheres. Sympathetic nerve stimulation caused significant reductions in blood flows in the facial tissues including the eye. Alpha-adrenoceptor blockade with phenoxybenzamine and combined beta- and alpha-adrenoceptor blockade with propranolol and phenoxybenzamine abolished the effects of sympathetic nerve stimulation in most facial tissues except in the tongue, upper eyelid and masseter muscle. In most cranial tissues, neuropeptide Y reduced regional blood flow and increased vascular resistance. No effect of neuropeptide Y on vascular resistance was observed in the choroid. In the present study, evidence for a non-adrenergic component in sympathetic vasoconstriction was found in the tongue, upper eyelid and masseter muscle but not in the majority of feline facial tissues. Neuropeptide Y was a potent vasoconstrictor in many cranial tissues, while in parts of the uvea, the effects of neuropeptide Y were less pronounced.     

Demonstration of a neuropeptide Y (NPY)-like immunoreactivity in the pigeon retina

The distribution of neuropeptide Y (NPY)-like immunoreactivity in the pigeon retina was investigated by fluorescence immunohistochemistry. NPY-positive cells were found in central and peripheral retina. NPY somata were located in the proximal portion of the inner nuclear layer and their processes directed to the inner plexiform layer where they ramified in 3 immunoreactive bands. NPY might play a role as a neurotransmitter or neuromodulator in the pigeon retina.     

Inhibitory effect of neuropeptide Y (NPY) on the in vitro activity of dopamine-beta-hydroxylase.

Neuropeptide Y (NPY) is found to be costored with norepinephrine (NE) in vesicles of the nerve terminals. An endogenous inhibitor of dopamine-beta-hydroxylase (DBH), the synthetic enzyme of NE, has been mentioned. In an attempt to clarify the effect of NPY on DBH activities, an in vitro assay is carried out using chromatographic analysis of NE formation from dopamine. NPY (20-80 pmol/ml) produced a dose-dependent depression of NE formation catalysed by the purified bovine adrenal DBH. Lineweaver-Burke plots (Km = 1.1 mM, Vmax = 10 pmol/min/mg protein) showed a non-competitive inhibition in NPY (30 pmol/ml, IC50)-treated samples. Moreover, failure of denatured NPY even at maximum concentration to influence the DBH activities suggested the essential of natural form of NPY. Participation of sulfhydryl compound seems also negligible, because N-ethylmaleimide did not overcome the effect of NPY. These results indicate that NPY has the ability to inhibit the catalytic action of DBH.     

Effects of 2-(2-Pyridyl)ethylamine (PEA) on the isolated guinea-pig heart

The effect of 2-(2-pyridyl)ethylamine (PEA) on the rate and force of contraction of cardiac tissues from untreated and reserpine-pretreated guinea-pigs was examined. PEA produced changes in rate of spontaneously beating right atria without activation of H₁ or H₂-receptors. The positive chronotropic effect of PEA was eliminated when atria from reserpine-pretreated animals were used, indicating an entirely indirect mode of action of PEA in the right atrium.The positive inotropic effect of low doses of PEA in left atria was antagonized by promethazine, whereas the inotropic effect of higher doses of PEA was reduced either by propranolol or following reserpine pretreatment. In the right ventricle strip, the inotropic effect of high doses of PEA was blocked equally by propranolol and promethazine.The results obtained indicate that in the guinea-pig heart, PEA either produces its effects through catecholamine release (right atrium) or through a combination of both H₁-receptor stimulation and catecholamine release (left atrium and right ventricle strip). The direct inotropic effects of PEA produced only small increases in force of contraction. The study indicates that caution should be used in ascribing all of the effects of PEA to histamine receptor stimulation.Supported by a grant from the Canadian Heart Foundation.Predoctoral Research Trainee of the Canadian Heart Foundation.     

Lithium inhibits internalization and endosomal processing of both neuropeptide Y (NPY) Y1 and transferrin receptors

Low concentrations of Li+ reduce the rate of internalization of neuropeptide Y (NPY) Y1 receptors [M.S. Parker, S.L. Parker, J.K. Kane, Internalization of neuropeptide Y Y1 and Y5 and of pancreatic polypeptide Y4 receptors is inhibited by lithium in preference to sodium and potassium ions, Regul. Pept., 118 (2004) 67-74]. This Li+-induced decrease in Y1 receptor internalization could be alleviated by Y1 receptor agonists. As shown by fractionation on Percoll gradients, lithium treatment induces a concentration-related decrease of intermediate and higher endosomal densities that contain the internalized Y1 ligand-receptor complex. This indicates an inhibition of endosome processing and maturation. Internalization of human transferrin shows [Li+] sensitivity similar to that of the Y1 receptor, and a similar Li+-induced decrease in endosomal processing. Lithium treatment thus decreases activity of the endosome system shared in the recycling endocytosis of the Y1 and transferrin receptors.     

A neuropeptide Y (NPY)-related peptide is present in the river lamprey CNS

Pancreatic polypeptide (PP)-like material from brain + spinal cord, and retina extracts of Lampetra fluviatilis was studied by HPLC and RIA. The brain + spinal cord extract showed a complex elution profile with multiple peaks of immunoreactivity. The retina extract showed a much simpler pattern with a single significant peak along with a trace of a second peak corresponding to the latest and penultimate peaks in the brain extract. Twenty-one out of 36 residues could be sequenced from the latest eluting peak in the brain extract. This sequence showed 81% identity with porcine neuropeptide (NPY) suggesting that both the brain/spinal cord and retina of the river lamprey contain a peptide homologous to NPY.     

Increased neuropeptide Y (NPY)-like immunoreactivity in rat sensory neurons following peripheral axotomy.

The effects of peripheral axotomy (sciatic nerve transection) on the presence and distribution of neuropeptide Y (NPY) in rat dorsal root ganglion (DRG) and spinal grey matter were examined using immunocytochemistry. In normal rats and on the sham-operated side of experimental rats, NPY-like immunoreactivity (NPYir) was observed in all laminae of the lumbar spinal cord, with an especially dense concentration of immunostained axons and axonal varicosities in laminae I-II of the dorsal horn. There was no detectable NPYir in L4-L5 DRG cells from normal rats or from the sham-operated side of experimental rats. At 14 days after axotomy, there was a large ipsilateral increase in the density of NPYir axons and varicosities in the lumbar spinal cord on the side of the nerve injury; this was especially apparent in laminae III-V. In the same rats, NPYir was observed in many small, medium, and large neurons in the L4-L5 DRGs on the side of the severed nerve.     

Opposite effects of centrally administered neuropeptide Y (NPY) on locomotor activity of spontaneously hypertensive (SH) and normal rats

Centrally administered neuropeptide Y has been shown to produce sedation manifested by a suppression of locomotor activity and a synchronizing effect on the EEG pattern in normal rats. It has been suggested that this sedative effect of NPY is largely due to a facilitation of the alpha 2-adrenergic transmission line. In the spontaneously hypertensive (SH) rat, the NPY-induced up-regulation of alpha 2-adrenoceptors observed in normal rats is absent, and NPY produces a desynchronization of the EEG. In the present study, we have therefore examined the effects of NPY on locomotor activity of SH rats and of inbred controls of the Wistar-Kyoto (WKy) strain, in both morning and evening sessions. In morning sessions, NPY (0.2-5.0 nmol intracerebroventricularly, i.c.v.) increased locomotor activity of SH rats in a dose-related manner. WKy rats were largely inactive per se, and no effects of NPY could be detected. In evening sessions, when spontaneous activity is high, NPY (I.0 nmol i.c.v.) still increased the activity of the SH rats. In WKy rats, an activity suppression similar to that previously reported for normal Sprague-Dawley rats was seen. The present results indicate that the sedative action of NPY in different rats strains correlates with the ability of the peptide to up-regulate alpha 2-adrenergic receptors.     

Occurrence of neuropeptide Y (NPY)-like immunoreactivity in catecholamine neurons in the human medulla oblongata

We report here the coexistence of a neuropeptide and catecholamines in neurons of the human brain. Using indirect immunofluorescence histochemistry, combined with elution and restaining experiments, neurons in the medulla oblongata of man were demonstrated to contain both a neuropeptide Y-like peptide and the catecholamine synthesizing enzyme tyrosine hydroxylase.     

Effects of neuropeptide Y on appetite

Neuropeptide Y (NPY) is a polypeptide containing 36 amino acids. Circulating NPY originates predominantly from the sympatho-adrenomedullary nervous system. It has a vasoconstrictive and mitogenic effect on blood vessels and seems to be involved in blood pressure regulation and angiogenesis. NPY is a potent orexigenic agent and is presumed to play a leading role in the regulation of eating behavior. Stimulation of the NPY-ergic arcuate - paraventricular nucleus (ARC-PVN) pathway by exercise, fasting, energy loss (glucosuria) is followed by increased appetite and food intake and increased parasympathetic activity, but suppression of sympathetic activity and energy expenditure. The end result of this process is an increase of energy stores. Activity of the NPY-ergic ARC-PVN pathway is suppressed by leptin - a polypeptide produced by adipocytes. Although functioning of an NPY-leptin feedback was found in rodents, it seems likely that also in man the NPY-leptin axis is involved in the regulation of food intake and energy expenditure.     

Hypothalamic neuropeptide Y (NPY) in obese Zucker rats: implications in feeding and sexual behaviors

Neuropeptide Y (NPY), a peptide of the pancreatic polypeptide family, is actually considered to be the most potent stimulator of food intake in rats when centrally injected. It has also suppressive effects on several components of sexual behavior. It was measured in discrete microdissected brain nuclei in obese hyperphagic Zucker fa/fa rats also characterized by a deficient reproductive function, as well as in their lean homozygous (Fa/Fa) and heterozygous (Fa/fa) counterparts. When compared with the lean (Fa/Fa) rats, NPY concentrations were significantly increased in the obese rats in the arcuate nucleus-median eminence (ARCME, +300%), in the paraventricular (PVN, +60%), suprachiasmatic (SCH, +90%), accumbens (+100%) and supraoptic (+40%) nuclei, as well as in the median preoptic area (MPOA, +70%). As PVN is one of the most important nuclei involved in the control of food intake and one site of NPY action, the high levels found in this nucleus might be a major component at the origin of hyperphagia in the obese animals. Food intake might be overstimulated by a sustained production of NPY as shown by the high concentrations found in the ARCME. NPY might also intervene in the pattern of food intake, for NPY contents were also largely modified in the SCH, the nucleus regulating feeding periodicity and in the MPOA, which is possibly involved in the regulation of energy balance. Finally, as the MPOA is the only site of action of NPY on sexual behavior, the higher levels measured in this area might contribute to the defective reproductive function of the obese Zucker fa/fa rat.     

Expression of phenylethanolamine N-methyltransferase (PNMT) and neuropeptide Y (NPY) in embryonic rat medulla oblongata grown in Oculo

Expression and development of specific markers of the adrenergic phenotype were studied in central neurons grown in transplant system. Medulla oblongata from embryonic day 12.5 (E12.5) or E18 rat was grafted into the anterior chamber of the eye of adult rat hosts. After two months, grafts were examined for the presence of immunoreactivity (IR) and catalytic activity to the epinephrine-synthesizing enzyme, phenylethanolamine N-methyltransferase (PNMT, E.C. 2.1.1.28), a specific adrenergic marker. In addition, grafts were examined for immunoreactivity to neuropeptide Y (NPY) and tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis. In E12.5 grafts, PNMT was expressed de novo, enzyme activity developed to levels similar to those in adult rat brainstem and PNMT-IR neurons were observed. TH-IR and NPY-IR neurons were also observed. In contrast, PNMT-IR was not observed in E18 grafts even though these already contained PNMT-IR neurons at the time of grafting. This was not due to poor growth of E18 grafts, in general, since TH-IR neurons were present and the protein content of the grafts was similar to that of E12.5 grafts. These studies suggest that adrenergic neurons survive well in oculo if they are transplanted prior to the age when neuroblasts have initially expressed the adrenergic phenotype, migrated to their final positions and elaborated processes. In addition, these studies establish a transplant system in which factors required for the development of central adrenergic neurons can be more easily studied than in situ.     

Subcellular storage and axonal transport of neuropeptide Y (NPY) in relation to catecholamines in the cat

The subcellular storage of neuropeptide Y-like immunoreactivity (NPY-LI) in peripheral sympathetic neurons and adrenal gland as well as its axonal transport in the sciatic nerve was studied in relation to catecholamines in the cat. In the subcellular fractions from different parts of sympathetic neurons, i.e. cell bodies (coeliac ganglia), axons (sciatic nerve) and terminals (spleen), the NPY-LI was found together with noradrenaline (NA) in heavy fractions assumed to contain large dense-cored vesicles. In addition, minor lighter fractions in the coeliac ganglion contained NPY-LI. The molar ratio between vesicular NA and NPY was high in the terminal regions (150 to 1) and much lower in axons and cell bodies (10 to 1), thus reflecting the different mechanisms of resupply for classical transmitter and peptide. In the adrenal gland the NPY-LI was mainly located in the catecholamine-storing chromaffin-granule fraction and also to a smaller extent in lighter fractions. Using reversed-phase HPLC, one molecular form of NPY-LI corresponding to porcine NPY was found in the coeliac ganglion, while the adrenal medulla also contained minor peaks with NPY-LI in addition to the main form, which co-eluted with porcine NPY. NA was stored both in light and heavy fractions in the spleen, while it was mainly found in heavier fractions in the sciatic nerve. In the coeliac ganglion, most of the noradrenaline was present in a non-particulate form. The anterograde transport rate for NPY-LI in the sciatic nerve was estimated to be about 9 mm h-1. A minor retrograde transport of NPY-LI was also detected. In conclusion, the present data suggest that NPY, a peptide with sympathoactive actions, is co-stored with NA in heavy fractions corresponding to large dense-cored vesicles, while light fractions with small dense-cored vesicles probably contain NA but not NPY-LI. The main resupply of NPY to terminals is, in contrast to NA, most likely by axonal transport, which implicates differences in the storage, turnover and release of these co-existing substances in the sympathoadrenal system.     

Neuropeptide Y (NPY) inhibits dimethylphenylpiperazinium (DMPP)-induced gastric acid secretion in isolated rat stomach

The effect of neuropeptide Y (NPY) on gastric acid secretion was investigated on an everted preparation of isolated rat stomach. Perfusion with synthetic NPY did not modify the basal secretion of gastric acid. However, NPY reduced dimethylphenylpiperazinium (DMPP)-stimulated acid secretion at concentrations insufficient to affect acid secretion stimulated by muscarine, histamine or gastrin. The decrease in acetylcholine (ACh) release from postganglionic cholinergic neurons by NPY is therefore considered to be responsible. Determination of NPY content by radioimmunoassay, in mucosal and muscular layers of the stomach, indicates that NPY possibly produces cholinergic inhibition under physiological levels. The present study suggests, therefore, that NPY has the ability to inhibit the release of ACh from postganglionic cholinergic neurons, thus producing a decrease in gastric acid secretion.     

Galanin and neuropeptide Y in chromaffin granules from the guinea-pig

We have examined the subcellular distribution of galanin-like immunoreactivity, neuropeptide Y-like immunoreactivity and the catecholamines noradrenaline and adrenaline in the adrenal medulla from guinea-pigs. By differential centrifugation of the adrenal medulla homogenate the neuropeptides as well as the catecholamines sedimented in a 10000 g pellet. This pellet was resuspended and further examined in discontinuous and continuous density gradients. In the discontinuous gradient the catecholamines peaked in the heavy bottom fraction, assumed to contain chromaffin granules. Galanin-like immunoreactivity and neuropeptide Y-like immunoreactivity were also enriched in this fraction. However, both neuropeptides showed high levels of sedimentable material also in a fraction of intermediate density. In the continuous density gradient, the sum of sedimentable and soluble catecholamines showed peak values in two fractions corresponding to 1.07 and 1.47 m sucrose, respectively. The NA peak in the denser fraction was more pronounced than the corresponding A peak. Galanin-like immunoreactivity showed only one peak, in the fraction corresponding to 1.07 M sucrose. The data suggest that galanin-like immunoreactivity and neuropeptide Y-like immunoreactivity are partly stored with catecholamines in chromaffin granules. However, galanin-like immunoreactivity and neuropeptide Y-like immunoreactivity was also found in fractions lighter than those containing the bulk of the catecholamines.     

Differential co-existence of neuropeptide Y (NPY)-like immunoreactivity with catecholamines in the central nervous system of the rat

The distribution of neuropeptide Y immunoreactive cell bodies in relation to various types of catecholamine-containing cell bodies in the rat brain was analyzed immunohistochemically using antisera to tyrosine hydroxylase, dopamine β-hydroxylase and phenylethanolamine N-methyltransferase. Coexistence of the peptide in catecholamine cell bodies was established by using an elution-restaining procedure.Neuropeptide Y-like immunoreactivity was observed in most noradrenergic cell bodies of the Al/Cl cell groups in the ventro lateral medulla oblongata. Similarly this peptide immunoreactivity was also observed in the majority of the adrenergic cell bodies of the C2 group. In the dorsal and dorsal-lateral part of the nucleus of the solitary tract, where a group of small adrenergic cells is present, several small neuropeptide Y immunoreactive cells were also observed. The possibility of coexistence of adrenaline and neuropeptide Y in these cells remains to be established. The majority of the noradrenergic cell bodies of the A2 group, as well as the presumptive dopaminergic cells within its ventromedial part, seemed to lack neuropeptide Y-like immunoreactivity. Many noradrenergic cell bodies of the A6 group in the locus coeruleus proper were neuropeptide Y-immunoreactive, whereas the peptide could not be observed in the subcoeruleus group. Neither the A5 and A7 noradrenergic cells in the pons, nor any of the dopaminergic cell groups in the mesencephalon and forebrain (A8–A15) seemed to contain a neuropeptide Y-like peptide.The findings indicate that central catecholamine neurons can be subdivided into distinct sub-groups based upon the coexistence of a specific peptide.