neuropeptide-y and Pain

Microglial cells are responsible for immune surveillance within the CNS. They respond to noxious stimuli by releasing inflammatory mediators and mounting an effective inflammatory response. This is followed by release of anti-inflammatory mediators and resolution of the inflammatory response. Alterations to this delicate process may lead to tissue damage, neuroinflammation, and neurodegeneration. Chronic pain, such as inflammatory or neuropathic pain, is accompanied by neuroimmune activation, and the role of glial cells in the initiation and maintenance of chronic pain has been the subject of increasing research over the last two decades. Neuropeptides are small amino acidic molecules with the ability to regulate neuronal activity and thereby affect various functions such as thermoregulation, reproductive behavior, food and water intake, and circadian rhythms. Neuropeptides can also affect inflammatory responses and pain sensitivity by modulating the activity of glial cells. The last decade has witnessed growing interest in the study of microglial activation and its modulation by neuropeptides in the hope of developing new therapeutics for treating neurodegenerative diseases and chronic pain. This review summarizes the current literature on the way in which several neuropeptides modulate microglial activity and response to tissue damage and how this modulation may affect pain sensitivity.

Topics: Adrenomedullin; Animals; Calcitonin Gene-Related Peptide; Ghrelin; Humans; Inflammation; Inflammation Mediators; Leptin; Macrophage Activation; Microglia; Neuralgia; Neurodegenerative Diseases; Neuroglia; Neuropeptide Y; Neuropeptides; Pain; Pro-Opiomelanocortin; Tachykinins; Vasoactive Intestinal Peptide

Topics: Animals; Central Nervous System Diseases; Emotions; Hippocampus; Humans; Neurodegenerative Diseases; Neuropeptide Y; Pain; Seizures; Stress, Psychological; Synapses

We review previously published data, and present some new data, indicating that spinal application of neuropeptide Y (NPY) reduces behavioral and neurophysiological signs of acute and chronic pain. In models of acute pain, early behavioral studies showed that spinal (intrathecal) administration of NPY and Y2 receptor agonists decrease thermal nociception. Subsequent neurophysiological studies indicated that Y2-mediated inhibition of excitatory neurotransmitter release from primary afferent terminals in the substantia gelatinosa may contribute to the antinociceptive actions of NPY. As with acute pain, NPY reduced behavioral signs of inflammatory pain such as mechanical allodynia and thermal hyperalgesia; however, receptor antagonist studies indicate an important contribution of spinal Y1 rather than Y2 receptors. Interestingly, Y1 agonists suppress inhibitory synaptic events in dorsal horn neurons (indeed, well known mu-opioid analgesic drugs produce similar cellular actions). To resolve the behavioral and neurophysiological data, we propose that NPY/Y1 inhibits the spinal release of inhibitory neurotransmitters (GABA and glycine) onto inhibitory neurons, e.g. disinhibition of pain inhibition, resulting in hyporeflexia. The above mechanisms of Y1- and Y2-mediated analgesia may also operate in the setting of peripheral nerve injury, and new data indicate that NPY dose-dependently inhibits behavioral signs of neuropathic pain. Indeed, neurophysiological studies indicate that Y2-mediated inhibition of Ca(2+) channel currents in dorsal root ganglion neurons is actually increased after axotomy. We conclude that spinal delivery of Y1 agonists may be of use in the treatment of chronic inflammatory pain, and that the use of Y1 and Y2 agonists in neuropathic pain warrants further consideration.

Topics: Analgesics; Animals; Disease Models, Animal; Injections, Spinal; Neuropeptide Y; Pain

Research during the past two decades supports a complex role for neuropeptide tyrosine (NPY) and two of its associated receptors, the Y1 receptor and the Y2 receptor, in the modulation of pain, in addition to regeneration and survival mechanisms at the spinal level. Thus, NPY has been shown to both cause and reduce pain, in addition to having biphasic effects. Recent research has focused on the distribution of the spinal NPY-mediated system. Here, we propose various possible scenarios for the role of NPY in pain processing, based on its actions at different sites (axon versus cell body), through different receptors (Y1 receptor versus Y2 receptor) and/or types of neuron (ganglion neurons and intraganglionic cross-excitation versus interneurons versus projection neurons).

Topics: Animals; Galanin; Ganglia, Spinal; Humans; Neuropeptide Y; Pain; Posterior Horn Cells; Spinal Cord

The expression of neuropeptide tyrosine (NPY) and two of its receptors (Y1- and Y2Rs) in different types of rodent dorsal root ganglion (DRG) and spinal cord neurons, and their regulation by peripheral nerve injury, have suggested a role in neuropathic pain. Here we present the spinal NPYergic system from an immunohistochemical perspective based on recent studies using two specific antibodies recognizing the Y1- and Y2Rs, respectively, as well as on data from a study on a Y1R knock-out mouse. We have, for example, defined seven different neuron populations of Y1R-expressing neurons in the rat spinal cord, representing multiple targets for spinally released NPY. The differential distribution of NPY receptors probably explains both the pro- and antinociceptive effects of NPY previously reported in the literature. One system possibly responsible for antinociception is a group of Y1R-positive, presumably glutamatergic interneurons in the superficial dorsal horn laminae. We also discuss the possibility that NPY released within DRGs can act in a paracrine fashion on NPY receptors on adjacent neurons, perhaps contributing to the so-called cross excitation, a concept advanced by Devor, Amir and collaborators. Taken together with behavioral and electrophysiological results summarized by Smith et al. in this volume, histochemical analyses have advanced the knowledge on the role of NPY in pain processing.

Topics: Animals; Neuropeptide Y; Pain; Pain Management; Receptors, Neuropeptide Y; Rodentia

Concepts related to the pathophysiology of reflex sympathetic dystrophy syndrome (RSDS) are changing. Although sympathetic influences are still viewed as the most likely mechanism underlying the development and/or perpetuation of RSDS, these influences are no longer ascribed to an increase in sympathetic tone. Rather, the most likely mechanism may be increased sensitivity to catecholamines due to sympathetic denervation with an increase in the number and/or sensitivity of peripheral axonal adrenoceptors. Several other pathophysiological mechanisms have been suggested, including neurogenic inflammation with the release of neuropeptides by primary nociceptive afferents and sympathetic efferents. These neuromediators, particularly substance P, calcitonin gene-related peptide, and neuropeptide Y (NPY), may play a pivotal role in the genesis of pain in RSDS. They induce an inflammatory response (cutaneous erythema and edema) and lower the pain threshold. Neurogenic inflammation at the site of the lesion with neuromediator accumulation or depletion probably contributes to the pathophysiology of RSDS. However, no single neuromediator has been proved responsible, and other hypotheses continue to arouse interest.

Topics: Calcitonin Gene-Related Peptide; Humans; Neurogenic Inflammation; Neuropeptide Y; Neuropeptides; Pain; Reflex Sympathetic Dystrophy; Substance P; Sympathetic Nervous System; Syndrome

Vasovagal syncope is defined as a reflex loss of consciousness related to reaction to various stimuli as orthostatic stress, pain or emotions connected with loss of muscle postural tone. The aetiology of this disorder is still unknown. The imbalance between the parts of autonomic nervous system and other homeostasis-related systems as renin-angiotensin-aldosterone system, peptides as endothelin, neuropeptide Y, vasopressin, adrenomedullin and cAMP, adenosine and AMP can play an important role in the development of vasovagal syncope. In the first part of the paper the authors describe the mechanisms involved in the development of vasovagal reaction, pathophysiology of the head-up tilt test and the role of autonomic nervous system.

Topics: Adrenomedullin; Cyclic AMP; Endothelins; Humans; Muscle Hypotonia; Neuropeptide Y; Pain; Peptides; Posture; Receptors, Neurotransmitter; Renin-Angiotensin System; Syncope, Vasovagal; Vasopressins

In the present brief overview we summarize results from several studies focusing on two neuropeptides, galanin and neuropeptide Y (NPY) in discrete neuronal systems, where they coexist with classic transmitters. On the basis of studies in different animal models we propose that these peptides may be involved in regulation of certain CNS functions and that drugs acting on their receptors may be of use in new therapeutic strategies. At the spinal level galanin and NPY are regulated in DRG neurons by nerve injury and in dorsal horn neurons by inflammation. It is possible that this leads to attenuation of pain sensitivity. Moreover, both peptides may exert trophic effects, for example to enhance regeneration. In the hypothalamic arcuate nucleus NPY and its receptors are part of the feeding circuitry, and we suggest that derangement of these NPY neurons may at least in part underlay the lethal phenotype of anorectic mice, which die 22 days postnatally after showing decreased food intake and growth retardation. Expression of NPY and NPY receptors is changed in the hippocampus of mice comparatively early after prion inoculation, indicating that this peptide system is affected in this spongiform degenerative disease in a region of importance for learning and memory. Finally, galanin is co-localized with classic monoamine transmitters in two central systems, the dorsal raphe serotonin neurons and the locus coeruleus noradrenergic neurons. In both cases galanin causes hyperpolarization (at high concentrations) and prolongs monoamine-induced outward currents (at low concentrations), thus modulating activity in two systems of importance for many brain functions including mood regulation. It may therefore be interesting to analyse to what extent drugs affecting galaninergic transmission also may be efficient in the treatment of, for example, depression.

Topics: Animals; Antidepressive Agents; Eating; Galanin; Humans; Nervous System Physiological Phenomena; Neuropeptide Y; Pain; Prion Diseases

The NPY neurons play an important role in information handling in the CNS by their ability to interact in both wiring and volume transmission at the network, local circuit and synaptic level. The importance of NPY/alpha 2 receptor-receptor interactions in cardiovascular, neuroendocrine and vigilance control is emphasized. Alterations in these receptor-receptor interactions take place in the spontaneously hypertensive rats as well as in the ischemic brain, which may have profound consequences for the information handling and contribute to the functional alterations found in these pathophysiological states. Finally, in the aging brain there appears to exist a marked reduction in NPY transmission line, which may affect higher brain functions, such as learning and memory retrieval. The most impressive result is, however, the indications of a role for NPY in volume transmission, where NPY appears to produce syndromic actions via its conversion into biologically active fragments, which may have preferential actions at Y2 NPY receptors. These syndromic pathways may be altered in the spontaneously hypertensive rat and may be controlled by gonadal steroids and glucocorticoids. Glucocorticoid receptors have been demonstrated in all arcuate NPY neurons and all NA/NPY and A/NPY costoring neurons.

Topics: Aging; Animals; Arousal; beta-Endorphin; Brain; Catecholamines; Hemodynamics; Ischemia; Neuropeptide Y; Pain; Rats; Rats, Inbred SHR; Receptors, Adrenergic, alpha; Receptors, Glucocorticoid; Synapses; Synaptic Membranes; Synaptic Transmission

The aim of this experiment was to measure the concentration of neuropeptide Y (NPY), a vasoactive transmitter which co-exists with noradrenaline in sympathetic nerve terminals, in venous blood taken from the painful and contralateral limbs of 16 patients with features of reflex sympathetic dystrophy (RSD) or causalgia. In nine patients tapping the skin of the affected limb provoked pain (allodynia). In seven of the nine patients with allodynia the concentration of NPY was lower on the painful side; similar results were obtained in only two of seven patients without widespread allodynia. In addition, the concentration of NPY was generally lower in the painful limb if it was warmer than the contralateral limb. These findings suggest that a reduction in sympathetic activity might accompany allodynia and influence vasomotor disturbances in patients with causalgic pain.

Topics: Adolescent; Adult; Arm; Causalgia; Female; Functional Laterality; Humans; Leg; Male; Middle Aged; Neuropeptide Y; Pain

Neuropeptide Y is associated with stress in animal and human laboratory studies. However, data from clinical studies are scarce and no clinical longitudinal studies have been published. The aim of this clinical study was to assess the possible association between changes in the levels of pain, depression, and stress measures, on the one hand, and plasma neuropeptide Y levels, on the other.. Forty-four women with the fibromyalgia syndrome were exposed to a Cognitive Behavioral Therapy intervention. Levels of the plasma neuropeptide Y as well as pain, depression, and stress measures were obtained at the start and at the end of the intervention, and after a further six month follow-up. Based on these data, a before-and-after analysis was performed.. Almost all measures of pain, depression, and stress improved during the study; specifically, variables measuring life control (coping), depression, and stress-related time urgency improved significantly. Moreover, during the same time period, the mean plasma neuropeptide Y level was reduced from 93.2 ± 38.8 fmol/mL before the Cognitive Behavioral Therapy to 75.6 ± 42.9 fmol/mL (p<0.001) at the end of the study.. After exposure to a Cognitive Behavioral Therapy intervention, levels of most of the pain, depression, and stress measures improved, half of them significantly, as did the levels of neuropeptide Y. This circumstance indicates a possible functional relationship between pain-depression-stress and neuropeptide Y.

Topics: Adaptation, Psychological; Female; Fibromyalgia; Humans; Longitudinal Studies; Neuropeptide Y; Pain

Topics: Humans; Hydrocortisone; Infant; Infant, Newborn; Neurokinin A; Neuropeptide Y; Pain; Prospective Studies; Substance P

Interoceptive sensing and hunger neurons have a role in the control of behavior.

Topics: Agouti-Related Protein; Animals; Behavior, Animal; Brain; Eating; Food Deprivation; Gastrointestinal Tract; Hunger; Mice; Neural Pathways; Neurons; Neuropeptide Y; Pain; Signal Transduction

Low doses of histamine or H1R agonist 2-pyridylethylamine (2-PEA) into the knee-joint were found to decrease formalin-induced articular nociception in rats. In this study, we evaluated the participation of spinal NPY in the antinociceptive effect produced by 2-PEA. Injection of formalin (1.5%) into one of the knee-joints causes the limping of the respective limb due to nociception, which was registered each 5 min over 60 min. Neuropeptide Y1 receptor (Y1R) content in the spinal cord was evaluated by western-blotting. Intrathecal (i.t.) injection of Y1R agonist Leu31, Pro34-NPY (0.7-7 µmol) decreased nociception, while injection of the antagonist BIBO 3304 (4 μmol), increased nociception. Antinociception produced by 2-PEA was reversed by a sub-effective i.t. dose of the Y1R antagonist. Similarly, this antinociceptive effect was prevented by i.t. pretreatment with the neurotoxin NPY-saporin (750 ng), which also reduced immunoblotting for Y1R in spinal cord homogenates. These data support the idea that antinociception induced by H1R agonists in the knee-joint of rats may be mediated by the spinal release of NPY, and this peptide seems to be acting via Y1R.

Topics: Analgesics; Animals; Arthralgia; Hindlimb; Injections, Intra-Articular; Injections, Spinal; Knee Joint; Male; Neuropeptide Y; Nociception; Pain; Pain Measurement; Pyridines; Rats; Rats, Wistar; Receptors, Histamine; Receptors, Neuropeptide Y; Spinal Cord; Spine

Transection of the sural and common peroneal branches of the sciatic nerve produces cutaneous hypersensitivity at the tibial innervation territory of the mouse hindpaw that resolves within a few weeks. We report that interruption of endogenous neuropeptide Y (NPY) signaling during remission, with either conditional NPY knockdown in NPY

Topics: Adenylyl Cyclases; Analgesics; Animals; Hyperalgesia; Male; Mice; Neuralgia; Neuropeptide Y; Nociception; Pain; Peripheral Nerve Injuries; Posterior Horn Cells; Receptors, N-Methyl-D-Aspartate; Receptors, Neuropeptide Y

Pain is a critical component hindering recovery and regaining of function after surgery, particularly in the elderly. Understanding the role of pain signaling after surgery may lead to novel interventions for common complications such as delirium and postoperative cognitive dysfunction. Using a model of tibial fracture with intramedullary pinning in male mice, associated with cognitive deficits, we characterized the effects on the primary somatosensory system. Here we show that tibial fracture with pinning triggers cold allodynia and up-regulates nerve injury and inflammatory markers in dorsal root ganglia (DRGs) and spinal cord up to 2 wk after intervention. At 72 h after surgery, there is an increase in activating transcription factor 3 (ATF3), the neuropeptides galanin and neuropeptide Y (NPY), brain-derived neurotrophic factor (BDNF), as well as neuroinflammatory markers including ionized calcium-binding adaptor molecule 1 (Iba1), glial fibrillary acidic protein (GFAP), and the fractalkine receptor CX3CR1 in DRGs. Using an established model of complete transection of the sciatic nerve for comparison, we observed similar but more pronounced changes in these markers. However, protein levels of BDNF remained elevated for a longer period after fracture. In the hippocampus, BDNF protein levels were increased, yet there were no changes in Bdnf mRNA in the parent granule cell bodies. Further, c-Fos was down-regulated in the hippocampus, together with a reduction in neurogenesis in the subgranular zone. Taken together, our results suggest that attenuated BDNF release and signaling in the dentate gyrus may account for cognitive and mental deficits sometimes observed after surgery.

Topics: Activating Transcription Factor 3; Animals; Brain-Derived Neurotrophic Factor; Calcium-Binding Proteins; Cognitive Dysfunction; CX3C Chemokine Receptor 1; Dentate Gyrus; Fracture Fixation, Intramedullary; Galanin; Ganglia, Spinal; Gene Expression Regulation; Glial Fibrillary Acidic Protein; Hyperalgesia; Male; Mice; Mice, Inbred C57BL; Microfilament Proteins; Neuropeptide Y; Pain; Proto-Oncogene Proteins c-fos; Sciatic Nerve; Signal Transduction; Spinal Cord; Tibial Fractures

5-hydroxytryptamine (5-HT) released in inflammatory tissues plays a pivotal role in pain hypersensitivity. However, it is not clear whether 5-HT2A receptors in the inflamed tissues mediate this effect. The present study investigated the contribution of 5-HT2A receptors in the periphery to chronic inflammatory pain. Complete Freund's adjuvant (CFA) was injected subcutaneously in the hindpaw of rats. The selective 5-HT2A receptor antagonist ketanserin was given in the inflamed site. Paw withdrawal latency responding to heat or mechanical stimuli was measured. Expression of neuropeptide Y (NPY) in the spinal dorsal horn and dorsal root ganglia (DRG) was assayed using immunohistochemistry technique. The results showed that ketanserin administered in the inflamed site inhibited thermal hyperalgesia in a dose-dependent manner (20, 40 and 80 µg) induced by the intraplantar injection of CFA. Ketanserin given once per day at a dose of 80 µg abolished heat hyperalgesia and also attenuated mechanical allodynia on the third day. CFA injection increased the expression of NPY in superficial laminae of the spinal cord, but not in the DRG. The local treatment of ketanserin completely inhibited CFA-induced increase in NPY expression in superficial laminae of the spinal cord. These results indicated that activation of 5-HT2A receptors in the inflamed tissues was involved in the pathogenesis of inflammatory pain and the blockade of 5-HT2A receptors in the periphery could relieve pain hypersensitivity and normalize the cellular disorder in the spinal dorsal horn associated with pathological pain. The present study suggests that the peripheral 5-HT2A receptors can be a promising target for pharmaceutical therapy to treat chronic inflammatory pain without central nervous system side effects.

Topics: Animals; Freund's Adjuvant; Ganglia, Spinal; Hot Temperature; Hyperalgesia; Inflammation; Ketanserin; Neuropeptide Y; Pain; Pain Measurement; Rats; Receptor, Serotonin, 5-HT2A; Serotonin; Serotonin 5-HT2 Receptor Antagonists; Spinal Cord Dorsal Horn

Smoking deregulates neuroendocrine responses to pain supporting production of neuropeptide Y (NpY) by direct stimulation of nicotinic receptors or by inhibiting adipokine leptin. Present study addressed the effect of cigarette smoking on adipokines and pain parameters, in 62 women with fibromyalgia (FM) pain syndrome with unknown etiology. Pain was characterized by a visual analogue scale, tender point (TP) counts, pressure pain threshold, and neuroendocrine markers NpY and substance P (sP). Levels of IGF-1, leptin, resistin, visfatin, and adiponectin were measured in blood and cerebrospinal fluid. Current smokers (n = 18) had lower levels of leptin compared to ex-smokers (n = 25, P = 0.002), while the expected NpY increase was absent in FM patients. In smokers, this was transcribed in higher VAS-pain (P = 0.04) and TP count (P = 0.03), lower pain threshold (P = 0.01), since NpY levels were directly related to the pain threshold (rho = 0.414) and inversely related to TP counts (rho = -0.375). This study shows that patients with FM have no increase of NpY levels in response to smoking despite the low levels of leptin. Deregulation of the balance between leptin and neuropeptide Y may be one of the essential mechanisms of chronic pain in FM.

Topics: Adult; Female; Fibromyalgia; Humans; Leptin; Middle Aged; Neuropeptide Y; Pain; Smoking

The precise etiology of knee osteoarthritis (KOA) pain remains highly controversial and there is no known effective treatment. Due to the known and suggested effects of neuropeptide Y (NPY) on pain, we have sought to investigate the relationship between the concentration of NPY in synovial fluid of knee, pain of KOA, and structural severity of KOA.. One hundred KOA patients and twenty healthy participants (control group) were recruited. The pain and the radiographic grade of KOA were assessed separately by Hideo Watanabe's pain score and Tomihisa Koshino's scoring system. Synovial fluid of knee from all participants was collected with arthrocentesis. Radioimmunoassay was used to examine the concentration of NPY in synovial fluid of knee.. Concentrations of NPY in synovial fluid were significantly higher in KOA patients (124.7 ± 33.4 pg/mL) compared with controls (64.8 ± 26.3 pg/mL) (p = 0.0297). According to Hideo Watanabe's pain score, 100 KOA patients were divided into 5 subgroups: no pain (n = 12), mild pain (n = 25), moderate pain (n = 37), strong pain (n = 19) and severe pain (n = 7). Within the KOA group, significantly higher concentrations of NPY were found in each subgroup as pain intensified (no pain 81.4 ± 11.7 pg/mL, mild pain 99.1 ± 23.2 pg/mL, moderate pain 119.9 ± 31.5 pg/mL, strong pain 171.2 ± 37.3 pg/mL and severe pain 197.3 ± 41.9 pg/mL). Meanwhile, according to Tomihisa Koshino's scoring system, 100 KOA patients were divided into 3 subgroups: early stage (n = 30), middle stage (n = 53), advanced stage (n = 17). Concentrations of NPY in middle and advanced stage groups of KOA patients were significant higher than early stage group of KOA patients (early stage 96.4 ± 27.1 pg/mL, middle stage 153.3 ± 16.9 pg/mL, advanced stage 149.5 ± 36.7 pg/mL) (p = 0.0163, p = 0.0352). Concentrations of NPY in advanced stage group of KOA patients has no significant difference compare with middle stage group of KOA patients (p = 0. 2175).. This study demonstrated the presence and variation of concentrations of NPY in the KOA joint fluid, suggesting a role for NPY as a putative regulator of pain transmission and perception in KOA pain.

Topics: Adult; Aged; Biomarkers; Female; Humans; Male; Middle Aged; Neuropeptide Y; Osteoarthritis, Knee; Pain; Pain Measurement; Synovial Fluid

Pain is a complex experience composed of sensory and affective components. Although the neural systems of the sensory component of pain have been studied extensively, those of its affective component remain to be determined. In the present study, we examined the effects of corticotropin-releasing factor (CRF) and neuropeptide Y (NPY) injected into the dorsolateral bed nucleus of the stria terminalis (dlBNST) on pain-induced aversion and nociceptive behaviors in rats to examine the roles of these peptides in affective and sensory components of pain, respectively. In vivo microdialysis showed that formalin-evoked pain enhanced the release of CRF in this brain region. Using a conditioned place aversion (CPA) test, we found that intra-dlBNST injection of a CRF1 or CRF2 receptor antagonist suppressed pain-induced aversion. Intra-dlBNST CRF injection induced CPA even in the absence of pain stimulation. On the other hand, intra-dlBNST NPY injection suppressed pain-induced aversion. Coadministration of NPY inhibited CRF-induced CPA. This inhibitory effect of NPY was blocked by coadministration of a Y1 or Y5 receptor antagonist. Furthermore, whole-cell patch-clamp electrophysiology in dlBNST slices revealed that CRF increased neuronal excitability specifically in type II dlBNST neurons, whereas NPY decreased it in these neurons. Excitatory effects of CRF on type II dlBNST neurons were suppressed by NPY. These results have uncovered some of the neuronal mechanisms underlying the affective component of pain by showing opposing roles of intra-dlBNST CRF and NPY in pain-induced aversion and opposing actions of these peptides on neuronal excitability converging on the same target, type II neurons, within the dlBNST.

Topics: Action Potentials; Affective Symptoms; Analysis of Variance; Aniline Compounds; Animals; Arginine; Corticotropin-Releasing Hormone; Cyclohexanes; Disease Models, Animal; Excitatory Amino Acid Antagonists; Formaldehyde; GABA Antagonists; Hormone Antagonists; Hormones; In Vitro Techniques; Kynurenic Acid; Male; Membrane Potentials; Microdialysis; Neurons; Neuropeptide Y; Pain; Pain Measurement; Peptide Fragments; Pyridazines; Pyrimidines; Rats; Rats, Sprague-Dawley; Septal Nuclei; Xanthenes

This study aimed to investigate temporal changes in glycine and its receptor expressions in cuneate neurons after median nerve transection (MNT), and the effects of glycine on neuropeptide Y (NPY) release and c-Fos expression in the cuneate nucleus (CN).. Immunohistochemistry methods were used to appraise changes of glycine- and GlyR-like immunoreactive (LI) neurons in the CN after MNT. The alterations in NPY and c-Fos expressions were used to assess the effects of saline, glycine or strychnine treatment. The CatWalk method was used to assess the efficiency of glycine treatment on the neuropathic signs of rats with MNT.. Approximately half of GlyR-LI neurons were fluorogold-labeled cuneothalamic projection neurons in the CN. Following MNT, the number of GlyR-LI neurons significantly decreased in the injured side of CN at 2 and 4 weeks, but the number of glycine-LI neurons remained unchanged. Four weeks after MNT given with electrical stimulation, strychnine significantly decreased the NPY reduction level in the stimulated side CN compared to that of the saline group. However, numbers of c-Fos-LI neurons in the glycine and strychnine groups were both significantly less than that in the saline group. But the paw print width and area in CatWalk analysis showed only a moderate recovery.. We conjecture that glycine increases glycine-mediated postsynaptic inhibition of cuneate neurons, and also blocks GABAergic neurons containing GlyRs which mediate presynaptic inhibition causing temperate NPY release. Consequently, the compromise results showed a weak reduction in c-Fos expression and a slight amelioration of neuropathic behaviors.

Topics: Animals; Gene Expression Regulation; Glycine; Immunohistochemistry; Median Nerve; Medulla Oblongata; Neuropeptide Y; Pain; Proto-Oncogene Proteins c-fos; Rats; Rats, Sprague-Dawley; Receptors, Glycine

The spinal Neuropeptide Y (NPY) system is a potential target for development of new pain therapeutics. NPY and two of its receptors (Y1 and Y2) are found in the superficial dorsal horn of the spinal cord, a key area of nociceptive gating and modulation. Lumbar intrathecal injection of (NPY) is antinociceptive, reducing hyper-reflexia to thermal and mechanical stimulation, particularly after nerve injury and inflammation. We have also shown that intrathecal injection of the targeted cytotoxin, Neuropeptide Y-sap (NPY-sap), is also antinociceptive, reducing nocifensive reflex responses to noxious heat and formalin. In the present study, we sought to determine the role of dorsal horn Y1R-expressing neurons in pain by destroying them with NPY-sap and testing the rats on three operant tasks. Lumbar intrathecal NPY-sap (1) reduced Complete Freund's Adjuvant (CFA)-induced hyper-reflexia on the 10°C cold plate, (2) reduced cold aversion on the thermal preference and escape tasks, (3) was analgesic to noxious heat on the escape task, (4) reduced the CFA-induced allodynia to cold temperatures experienced on the thermal preference, feeding interference, and escape tasks, and (5) did not inhibit or interfere with morphine analgesia.

Topics: Analgesics; Animals; Behavior, Animal; Cold Temperature; Female; Freund's Adjuvant; Hyperalgesia; Inflammation; Morphine; Neuropeptide Y; Pain; Posterior Horn Cells; Rats; Rats, Long-Evans; Receptors, Neuropeptide Y; Reflex; Spinal Cord

Despite recent progress in describing the common neural circuitry of emotion and stress processing, the bases of individual variation are less well understood. Genetic variants that underlie psychiatric disease have proven particularly difficult to elucidate. Functional genetic variation of neuropeptide Y (NPY) was recently identified as a source of individual differences in emotion. Low NPY levels have been reported in major depressive disorder (MDD).. To determine whether low-expression NPY genotypes are associated with negative emotional processing at 3 levels of analysis.. Cross-sectional, case-control study.. Academic medical center.. Among 44 individuals with MDD and 137 healthy controls, 152 (84%) had an NPY genotype classified as low, intermediate, or high expression according to previously established haplotype-based expression data.. Healthy subjects participated in functional magnetic resonance imaging while viewing negative (vs neutral) words (n = 58) and rated positive and negative affect during a pain-stress challenge (n = 78). Genotype distribution was compared between 113 control subjects and 39 subjects with MDD.. Among healthy individuals, negatively valenced words activated the medial prefrontal cortex. Activation within this region was inversely related to genotype-predicted NPY expression (P = .03). Whole-brain regression of responses to negative words showed that the rostral anterior cingulate cortex activated in the low-expression group and deactivated in the high-expression group (P < .05). During the stress challenge, individuals with low-expression NPY genotypes reported more negative affective experience before and after pain (P = .002). Low-expression NPY genotypes were overrepresented in subjects with MDD after controlling for age and sex (P = .004). Population stratification did not account for the results.. These findings support a model in which NPY genetic variation predisposes certain individuals to low NPY expression, thereby increasing neural responsivity to negative stimuli within key affective circuit elements, including the medial prefrontal and anterior cingulate cortices. These genetically influenced neural response patterns appear to mediate risk for some forms of MDD.

Topics: Adult; Amygdala; Arousal; Brain; Brain Mapping; Case-Control Studies; Cross-Sectional Studies; Depressive Disorder, Major; Emotions; Female; Gene Expression; Genetic Predisposition to Disease; Genetic Variation; Genotype; Gyrus Cinguli; Haplotypes; Humans; Magnetic Resonance Imaging; Male; Middle Aged; Nerve Net; Neuropeptide Y; Pain; Phenotype; Polymorphism, Single Nucleotide; Prefrontal Cortex; Reference Values; Young Adult

Dramatically up-regulated in the dorsal horn of the mammalian spinal cord following inflammation or nerve injury, neuropeptide Y (NPY) is poised to regulate the transmission of sensory signals. We found that doxycycline-induced conditional in vivo (Npy(tet/tet)) knockdown of NPY produced rapid, reversible, and repeatable increases in the intensity and duration of tactile and thermal hypersensitivity. Remarkably, when allowed to resolve for several weeks, behavioral hypersensitivity could be dramatically reinstated with NPY knockdown or intrathecal administration of Y1 or Y2 receptor antagonists. In addition, Y2 antagonism increased dorsal horn expression of Fos and phosphorylated form of extracellular signal-related kinase. Taken together, these data establish spinal NPY receptor systems as an endogenous braking mechanism that exerts a tonic, long-lasting, broad-spectrum inhibitory control of spinal nociceptive transmission, thus impeding the transition from acute to chronic pain. NPY and its receptors appear to be part of a mechanism whereby mammals naturally recover from the hyperalgesia associated with inflammation or nerve injury.

Topics: Animals; Arginine; Behavior, Animal; Benzazepines; Chronic Disease; Gene Expression Regulation; Mice; Mice, Knockout; Neuropeptide Y; Nociceptors; Pain; Posterior Horn Cells; Proto-Oncogene Proteins c-fos; Receptors, Neuropeptide Y; Synaptic Transmission

Chronic inflammation associated with osteoarthritis (OA) alters normal responses and modifies the functionality of the articular vasculature. Altered responsiveness of the vasculature may be due to excessive neural activity associated with chronic pain and inflammation, or from the production of inflammatory mediators which induce vasodilation. Using laser speckle perfusion imaging (LSPI), blood flow to the medial collateral ligament (MCL) of adult rabbits was measured in denervated ACL transected knees (n = 6) and compared to unoperated control (n = 6) and 6-week anterial cruciate ligament (ACL)-transected knees (n = 6). Phenylephrine and neuropeptide Y were applied to the MCL vasculature in topical boluses of 100 microL (dose range 10(-14) to 10(-8) mol and 10(-14) to 10(-9) mol, respectively). Denervation diminished vasoconstrictive responsiveness to phenylephrine compared to both control and ACL-transected knees. Denervation minimally enhanced vascular responses to neuropeptide Y (NPY) compared to ACL deficiency alone, which nevertheless remained significantly diminished from control responses. To evaluate the potential role of inflammatory dilators in the diminished contractile responses, phenylephrine was coadministered with histamine, substance P, and prostaglandin E(2). High-dose histamine, and low-dose substance P and PGE(2) were able to inhibit contractile responses in the MCL of control knees. Excessive neural input does not mediate diminished vasoconstrictive responses in the ACL transected knee; inflammatory mediators may play a role in the deficient vascular responsiveness of the ACL transected knee.

Topics: Adrenergic alpha-Agonists; Animals; Anterior Cruciate Ligament Injuries; Denervation; Disease Models, Animal; Dose-Response Relationship, Drug; Inflammation; Inflammation Mediators; Medial Collateral Ligament, Knee; Neuropeptide Y; Osteoarthritis, Knee; Pain; Phenylephrine; Physical Stimulation; Rabbits; Regional Blood Flow; Vasoconstriction

We used a new computer-assisted method to precisely localize and efficiently quantify increases in neuropeptide Y immunoreactivity (NPY-ir) along the mediolateral axis of the L4 dorsal horn (DH) following transection of either the tibial and common peroneal nerves (thus sparing the sural branch, spared nerve injury (SNI)), the tibial nerve, or the common peroneal and sural nerves. Two weeks after SNI, NPY-ir increased within the tibial and peroneal innervation territories; however, NPY-ir in the central-lateral region (innervated by the spared sural nerve) was indistinguishable from that of sham. Conversely, transection of the sural and common peroneal nerves induced an increase in NPY-ir in the central-lateral region, while leaving the medial region (innervated by the tibial nerve) unaffected. All nerve injuries increased NPY-ir in dorsal root ganglia (DRG) and nucleus gracilis (NG). By 24 weeks, both NPY-ir upregulation in the DH and hyper-responsivity to cold and noxious mechanical stimuli had resolved. Conversely, NPY-ir in DRG and NG, and hypersensitivity to non-noxious static mechanical stimuli, did not resolve within 24 weeks. Over this time course, the average cross-sectional area of NPY-immunoreactive DRG neurons increased by 151 mum(2). We conclude that the upregulation of NPY after SNI is restricted to medial zones of the DH, and therefore cannot act directly upon synapses within the more lateral (sural) zones to control sural nerve hypersensitivity. Instead, we suggest that NPY in the medial DH tonically inhibits hypersensitivity by interrupting mechanisms of central sensitization and integration of sensory signals at the spinal and supraspinal levels.

Topics: Animals; Cold Temperature; Ganglia, Spinal; Lumbar Vertebrae; Male; Medulla Oblongata; Neurons; Neuropeptide Y; Pain; Peroneal Nerve; Physical Stimulation; Posterior Horn Cells; Rats; Rats, Sprague-Dawley; Sciatic Nerve; Spinal Cord; Sural Nerve; Tibial Nerve; Time Factors

Fibromyalgia (FM) is thought to occur because of the combination of interactions among neurotransmitters, such as neuropeptide Y (NPY), stressors, hormones, cytokines, and both the immune and sympathetic nervous systems. The aim of this study was to evaluate serum concentrations of cytokines, antipolymer antibodies (APA), and NPY in 51 patients with FM, 25 with tension-type headache (TTH), and 15 healthy controls. Serum concentrations of eight different cytokines, APA and NPY, were measured. Interleukin (IL)-1RA, IL-6, IL-10, and tumor necrosis factor-alpha were higher in serum of FM patients compared with TTH patients and a significant correlation between IL-10 and Fibromyalgia Impact Questionnaire score was observed. There was a significant difference between FM and TTH versus controls in NPY levels, but not in APA levels. Cytokines and NPY take part in pain modulation and even if they are altered in FM they cannot be considered as measurable biomarkers of disease.

Topics: Adult; Aged; Antibodies; Case-Control Studies; Cytokines; Female; Fibromyalgia; Humans; Interviews as Topic; Male; Middle Aged; Neuropeptide Y; Pain; Surveys and Questionnaires; Syndrome; Tension-Type Headache

GPR103 is one of the orphan G protein-coupled receptors. Recently, an endogenous ligand for GPR103, 26RFa, was identified. Many 26RFa binding sites have been observed in various nuclei of the brain involved in the processing of pain such as the parafascicular thalamic nucleus, the locus coeruleus, the dorsal raphe nucleus, and the parabrachial nucleus. In the present study, the effects of intracerebroventricular injection of 26RFa were tested in the rat. Intracerebroventricular injection of 26RFa significantly decreased the number of both phase 1 and phase 2 agitation behaviors induced by paw formalin injection. This analgesic effect of 26RFa on the phase 1 response, but not phase 2 response, was antagonized by BIBP3226, a mixed antagonist of neuropeptide Y Y1 and neuropeptide FF receptors. Intracerebroventricular injection of 26RFa has no effect in the 52.5 degrees C hot plate test. Intracerebroventricular injection of 26RFa had no effect on the expression of Fos-like immunoreactivity induced by paw formalin injection in the superficial layers of the spinal dorsal horn. These data suggest that (1) 26RFa modulates nociceptive transmission at the supraspinal site during a formalin test, (2) the mechanism 26RFa uses to produce an analgesic effect on the phase 1 response is different from that on the phase 2 response, and (3) intracerebroventricularly injected 26RFa dose not directly inhibit the nociceptive input to the spinal cord.

Topics: Analgesics, Non-Narcotic; Animals; Anti-Anxiety Agents; Arginine; Formaldehyde; Hot Temperature; Injections, Intraventricular; Male; Narcotic Antagonists; Neuropeptide Y; Neuropeptides; Oligopeptides; Pain; Pain Measurement; Proto-Oncogene Proteins c-fos; Rats; Rats, Sprague-Dawley; Reaction Time; Receptors, G-Protein-Coupled; Spinal Cord

Topics: Animals; Comorbidity; Depressive Disorder; Disease Models, Animal; Gene Expression Regulation; Humans; Neuropeptide Y; Pain; Pain Management; Peripheral Nervous System Diseases; Rats; Reproducibility of Results; Research Design; Synaptic Transmission; Translational Research, Biomedical

cis-4-(Piperazin-1-yl)-5,6,7a,8,9,10,11,11a-octahydrobenzofuro[2,3-h]quinazolin-2-amine, 4 (A-987306) is a new histamine H(4) antagonist. The compound is potent in H(4) receptor binding assays (rat H(4), K(i) = 3.4 nM, human H(4) K(i) = 5.8 nM) and demonstrated potent functional antagonism in vitro at human, rat, and mouse H(4) receptors in cell-based FLIPR assays. Compound 4 also demonstrated H(4) antagonism in vivo in mice, blocking H(4)-agonist induced scratch responses, and showed anti-inflammatory activity in mice in a peritonitis model. Most interesting was the high potency and efficacy of this compound in blocking pain responses, where it showed an ED(50) of 42 mumol/kg (ip) in a rat post-carrageenan thermal hyperalgesia model of inflammatory pain.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Benzofurans; Carrageenan; Disease Models, Animal; Drug Design; Drug Evaluation, Preclinical; Humans; Hyperalgesia; Ligands; Mice; Molecular Structure; Pain; Peritonitis; Quinazolines; Rats; Receptors, G-Protein-Coupled; Receptors, Histamine; Receptors, Histamine H4; Stereoisomerism; Structure-Activity Relationship

Recent pharmacologic studies in our laboratory have suggested that the spinal neuropeptide Y (NPY) Y1 receptor contributes to pain inhibition and to the analgesic effects of NPY. To rule out off-target effects, the present study used Y1-receptor-deficient (-/-) mice to further explore the contribution of Y1 receptors to pain modulation.. Y1(-/-) mice exhibited reduced latency in the hotplate test of acute pain and a longer-lasting heat allodynia in the complete Freund's adjuvant (CFA) model of inflammatory pain. Y1 deletion did not change CFA-induced inflammation. Upon targeting the spinal NPY systems with intrathecal drug delivery, NPY reduced tactile and heat allodynia in the CFA model and the partial sciatic nerve ligation model of neuropathic pain. Importantly, we show for the first time that NPY does not exert these anti-allodynic effects in Y1(-/-) mice. Furthermore, in nerve-injured CD1 mice, concomitant injection of the potent Y1 antagonist BIBO3304 prevented the anti-allodynic actions of NPY. Neither NPY nor BIBO3304 altered performance on the Rotorod test, arguing against an indirect effect of motor function.. The Y1 receptor contributes to pain inhibition and to the analgesic effects of NPY.

Topics: Analgesics; Animals; Anti-Inflammatory Agents; Arginine; Behavior, Animal; Disease Models, Animal; Hot Temperature; Hyperalgesia; Inflammation; Male; Mice; Mice, Knockout; Neuropeptide Y; Pain; Pain Measurement; Receptors, Neuropeptide Y; Sciatic Neuropathy

Understanding inter-individual differences in stress response requires the explanation of genetic influences at multiple phenotypic levels, including complex behaviours and the metabolic responses of brain regions to emotional stimuli. Neuropeptide Y (NPY) is anxiolytic and its release is induced by stress. NPY is abundantly expressed in regions of the limbic system that are implicated in arousal and in the assignment of emotional valences to stimuli and memories. Here we show that haplotype-driven NPY expression predicts brain responses to emotional and stress challenges and also inversely correlates with trait anxiety. NPY haplotypes predicted levels of NPY messenger RNA in post-mortem brain and lymphoblasts, and levels of plasma NPY. Lower haplotype-driven NPY expression predicted higher emotion-induced activation of the amygdala, as well as diminished resiliency as assessed by pain/stress-induced activations of endogenous opioid neurotransmission in various brain regions. A single nucleotide polymorphism (SNP rs16147) located in the promoter region alters NPY expression in vitro and seems to account for more than half of the variation in expression in vivo. These convergent findings are consistent with the function of NPY as an anxiolytic peptide and help to explain inter-individual variation in resiliency to stress, a risk factor for many diseases.

Topics: Alleles; Anxiety; Anxiety Disorders; Brain; Emotions; Facial Expression; Finland; Gene Expression Regulation; Genetic Variation; Haplotypes; Humans; Lymphocytes; Magnetic Resonance Imaging; Male; Neuropeptide Y; Opioid Peptides; Pain; Polymorphism, Single Nucleotide; RNA, Messenger; Stress, Physiological; United States; White People

Neuropeptide Y (NPY) induces analgesic-like effects after central administration across diverse pain models in rodents. In spinal pain models, previous studies indicate a prominent role for Y(1) receptors at mediating this effect of NPY. In supraspinal pain models like the hot plate test, the NPY receptors involved have not been thoroughly explored. By intracerebroventricular (i.c.v.) administration of selective NPY receptor ligands, the possible involvement of Y(5) receptors in analgesic-like mechanisms was investigated using the hot plate test in rats. Both NPY and selective Y(5) agonists induced analgesic-like effects as revealed by prolonged hot plate latencies. Further consistent with a role for Y(5) receptors, pretreatment with a selective Y(5) receptor antagonist blocked the Y(5) agonist-induced analgesic-like effect. The present study indicates involvement of Y(5) receptors probably at the supraspinal level in mediation of NPY agonist-induced analgesic-like effects in the hot plate test.

Topics: Analgesics; Animals; Disease Models, Animal; Hot Temperature; Injections, Intraventricular; Neuropeptide Y; Pain; Peptide Fragments; Rats; Receptors, Neuropeptide Y

In this study, we examined characteristics of the neuropeptide Y-like immunoreactive (NPY-LI) dorsal root ganglion (DRG) neurons after complete median nerve transection (CMNT). With fluorogold (FG) injection into normal median nerves, numerous FG-labeled DRG neurons were localized predominantly in the C6 and C7 DRGs, where the focal regions were examined after CMNT. With NPY immunohistochemistry, a few NPY-LI neurons were detected in the ipsilateral but not contralateral DRGs after FG injection into the nerve. As early as 3 days after CMNT a few NPY-LI neurons could be detected, reaching a maximum in the DRGs at 4 weeks, subsiding thereafter over 20 weeks. The NPY-LI DRG neurons were primarily medium-sized and large neurons. With FG injection into the transected median nerve, we found that approximately 99% of NPY-LI neurons were labeled for FG, suggesting that they were derived from the injured but not intact DRG neurons. Using double fluorescent dyes tracing, we detected that some of the injured DRG neurons were NPY-LI neurons that projected to the cuneate nucleus (CN). Following dorsal rhizotomy, our data indicated that after CMNT the induced NPY-LI fibers in the ipsilateral CN originated exclusively from the injured DRG neurons. Taken together, these findings suggest that injury-induced NPY-LI fibers in the CN may originate from the injured DRG neurons via the median primary afferent fibers, affect the excitability of cuneothalamic projection neurons (CTNs), and involve neuropathic sensation following CMNT.

Topics: Animals; Axotomy; Fluorescent Dyes; Ganglia, Spinal; Immunohistochemistry; Male; Median Nerve; Medulla Oblongata; Neural Pathways; Neurons, Afferent; Neuropeptide Y; Pain; Rats; Rats, Wistar

The localization of the neuropeptide tyrosine Y1 receptor was studied with immunohistochemistry in parasagittal and transverse, free-floating sections of the rat lumbar spinal cord. At least seven distinct Y1 receptor-positive populations could tentatively be recognized: Type 1) abundant small, fusiform Y1 receptor-positive neurons in laminae I-II, producing a profuse neuropil; Type 2) Y1 receptor-positive projection neurons in lamina I; Type 3) small Y1 receptor-positive neurons in lamina III, similar to Type 1 neurons, but less densely packed; Type 4) a number of large, multipolar Y1 receptor-positive neurons in the border area between laminae III-IV, with dendrites projecting toward laminae I-II; Type 5) a considerable number of large, multipolar Y1 receptor-positive neurons in laminae V-VI; Type 6) many large Y1 receptor-positive neurons around the central canal (area X); and Type 7) a small number of large Y1 receptor-positive neurons in the medial aspect of the ventral horns (lamina VIII). Many of the neurons present in laminae V-VI and area X produce craniocaudal processes extending for several hundred micrometers. Retrograde tracing using cholera toxin B subunit injected at the 9th thoracic spinal cord level shows that several Type 5 neurons in laminae V-VI, and at least a few Type 2 in lamina I and Type 6 in area X have projections extending to the lower segments of the thoracic spinal cord (and perhaps to supraspinal levels). The present results define distinct subpopulations of neuropeptide tyrosine-sensitive neurons, localized in superficial and deep layers of the dorsal, in the ventral horns and in area X. The lamina II neurons express somatostatin [The neuropeptide Y Y1 receptor is a somatic receptor on dorsal root ganglion neurons and a postsynaptic receptor on somatostatin dorsal horn neurons. Eur J Neurosci 11:2211-2225] and are presumably glutamatergic [Todd AJ, Hughes DI, Polgar E, Nagy GG, Mackie M, Ottersen OP, Maxwell DJ (2003) The expression of vesicular glutamate transporters VGLUT1 and VGLUT2 in neurochemically defined axonal populations in the rat spinal cord with emphasis on the dorsal horn. Eur J Neurosci 17:13-27], that is they are excitatory interneurons under a Y1 receptor-mediated inhibitory influence. The remaining Y1 receptor-positive spinal neurons need to be phenotyped, for example if the large Y1 receptor-positive laminae III-IV neurons (Type 5) are identical to the neurokinin (NK)1R-positive neurons previously shown t

Topics: Animals; Calcitonin Gene-Related Peptide; Cell Shape; Cholera Toxin; Dendrites; Immunohistochemistry; Interneurons; Male; Neural Pathways; Neuropeptide Y; Neuropeptides; Neuropil; Neurotransmitter Agents; Pain; Posterior Horn Cells; Presynaptic Terminals; Rats; Rats, Sprague-Dawley; Receptors, Neuropeptide Y; Tyrosine

To detect neuropeptides in human skeletal muscle at rest and after eccentric exercise.. Eight healthy subjects participated in the study. Microdialysis of the distal part of the vastus lateralis of the quadriceps muscle and pain evaluation were performed immediately after eccentric exercise, after two days, and at rest. Calcitonin gene related peptide (CGRP) and neuropeptide Y (NPY), representatives of the sensory and autonomic nervous system, were analysed by radioimmunoassay.. Overall, the measured concentrations were low, some even below the limit of detection. At rest, CGRP was detected in two of seven samples, but after eccentric exercise it was detected in 27 of 30 samples. At rest, all NPY concentrations were below the limit of detection, but after exercise it was found in six of 30 samples.. The significant increase in detectability of CGRP after eccentric exercise may be related to the increased experience of pain. Therefore the occurrence of CGRP after heavy eccentric exercise may be associated with the regulation of delayed onset muscle soreness and possibly also the stimulation of tissue regeneration.

Topics: Adult; Calcitonin Gene-Related Peptide; Exercise; Female; Humans; Male; Microdialysis; Muscle, Skeletal; Neuropeptide Y; Pain; Pain Measurement; Radioimmunoassay

The release of neuropeptide Y (NPY) is reported to increase in ischemic conditions and may thus be involved in chronic myalgia. The purpose of this study was to investigate the effect of isometric contraction on intramuscular levels of NPY in relation to local pain development.. Intramuscular microdialysis was performed in the masseter and trapezius muscles to determine NPY levels before, during, and after isometric contraction in 16 healthy females. Pain intensity was assessed simultaneously with VAS. Repeated measures ANOVA, t-test, and Pearson correlation analysis were used for statistical analyses.. The level of NPY in the trapezius muscle was increased during and after contraction, while there was no change in the masseter muscle. The level of NPY before contraction was higher in the masseter muscle than in the trapezius muscle, and the levels in the two muscles were correlated before and during contraction. Low-level pain in both muscles after probe insertion increased significantly during contraction, but the pain was not correlated to the NPY level.. Pain is developed in the trapezius and masseter muscles during repeated isometric contraction. The NPY level is increased in the trapezius muscle but is not associated with the pain development.

Topics: Bite Force; Female; Humans; Isometric Contraction; Masseter Muscle; Microdialysis; Middle Aged; Muscle, Skeletal; Neuropeptide Y; Pain; Pain Measurement; Pain Threshold

Neuropeptide Y (NPY) plays an important role in pain modulation at different levels in the central nervous system. In the brain, NPY and NPY receptors distribute abundantly in the arcuate nucleus of hypothalamus (ARC), a structure involved in pain processing. The present study was undertaken to investigate the role of NPY in nociceptive modulation in the ARC of intact rats and rats with carrageenan-induced inflammation. Intra-ARC administration of NPY induced dose-dependent increases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation in intact rats, which was attenuated by the Y1 receptor antagonist NPY28-36. Intra-ARC administration of NPY also induced dose-dependent increases in HWLs to noxious stimulation in rats with inflammation. Furthermore, intra-ARC injection of either the antiserum against NPY or NPY28-36 induced decreases in HWLs in rats with inflammation, while both of them produced no effects in intact ones. Additionally, there were marked increases of Y1 receptor in the bilateral ARC of rats with inflammation tested by immunohistochemistry, while no significant changes of NPY were observed, implicating that the increased Y1 receptor has an important effect in the NPY-induced antinociception. We also found that intra-ARC injection of Y2 receptor agonist NPY3-36 produced no significant antinociception in either intact rats or rats with inflammation. Together, we demonstrate that NPY exerts an antinociceptive effect in the ARC of intact rats and rats with inflammation. Both Y1 receptor and endogenous released NPY in the ARC are involved in the nociceptive modulation during inflammation.

Topics: Animals; Arcuate Nucleus of Hypothalamus; Behavior, Animal; Carrageenan; Hindlimb; Hyperalgesia; Immunohistochemistry; Inflammation; Male; Neuropeptide Y; Nociceptors; Pain; Pain Measurement; Peptide Fragments; Rats; Rats, Wistar; Reaction Time; Receptors, Neuropeptide Y

In order to clarify the physiologic role of NPY in sensory processing, we obtained intracellular recordings of DRG neurons from wild type (WT) and NPY overexpressing transgenic rats (NPY-TG) before and after injury. We investigated medium and large diameter DRG neurons since upregulation of NPY peptide following the nerve injury occurs primarily in those cells. Neurons were classified as Aalpha/beta and Adelta using conduction velocity and action potential duration. Prior to the injury, Aalpha/beta neurons of NPY-TG rats conducted more slowly and had a more brief AHP than similar cells from the WT group. Adelta neurons at baseline conducted faster in TG animals compared to WT. Ligation of the 5th lumbar spinal nerve (SNL) produced certain changes in Aalpha/beta cells that were evident only in the TG group. These include increased refractory period, increased input resistance, AHP prolongation and a depolarizing shift in threshold for AP initiation. The expected injury-induced CV slowing was not seen in NPY-TG Aalpha/beta cells. In the Adelta cell group, injury produced a depolarizing shift in the resting membrane potential, an increase in AP duration and decrease in AHP and refractory period duration only in WT rats, while NPY-TG cells lacked these injury-induced changes. Behavior tests showed diminished sensory response to nerve injury in NPY-TG rats, i.e., shorter duration of enhanced pain-related behavior and attenuation of contralateral effect. In conclusion, our observations suggest that NPY overexpression leads to reduced neuronal activity following nerve injury in a cell-specific manner.

Topics: Animals; Animals, Genetically Modified; Axotomy; Electrophysiology; Ganglia, Spinal; Immunohistochemistry; Motor Activity; Neurons, Afferent; Neuropeptide Y; Pain; Pain Threshold; Patch-Clamp Techniques; Rats; Rats, Sprague-Dawley; Spinal Nerves

Microdialysis of neuropeptide Y (NPY) has been reported to be difficult, which partly may be due to the adhesive nature of the molecule. The aim of this study was to determine the optimal probe and perfusion medium for microdialysis of NPY and to investigate if microdialysis can be used to sample NPY from human muscle tissue. Three different probe types with a 10mm membrane were used for experiments in vitro. They were perfused with a modified Krebs-Henseleit buffer (KHB), with or without the addition of 0.5% human serum albumin (HSA). Dialysate samples were collected at different flow rates. Ten healthy subjects participated in the clinical microdialysis. Microdialysis samples were obtained by probes inserted intramuscularly in the right masseter and trapezius muscle and perfused with KHB+HSA at a flow-rate of 4 microl/min. The relative recovery of NPY was significantly higher in the dialysates with HSA added to the perfusion medium, while there was no difference between the three probe types. NPY was detectable in 90% of the dialysates from the masseter and in 40% of the dialysates from the trapezius muscle. In conclusion, this study shows that NPY can be detected in microdialysis samples from human skeletal muscle and that addition of HSA to the perfusion medium increases the relative recovery in vitro.

Topics: Adult; Albumins; Buffers; Electrodes; Extracellular Fluid; Female; Humans; Masseter Muscle; Microcirculation; Microdialysis; Muscle, Skeletal; Neuropeptide Y; Pain; Perfusion

To test the hypothesis that administration of neuropeptide Y (NPY) to the spinal cord reduces inflammatory pain, we evaluated the effects of intrathecal NPY on behavioral and cardiovascular markers of the nociception associated with intraplantar formalin injection in rats. Before the administration of formalin, NPY dose dependently increased blood pressure, an effect that could be prevented with the coadministration of the Y2 antagonist, BIIE0246. This effect lasted only 20 min, and thus was over before initiation of the formalin test. NPY dose dependently inhibited the flinching, licking, pressor, and tachycardia responses associated with formalin injection. The Y1 receptor antagonist BIBO 3304 partially reversed the antinociceptive effect of NPY at a dose that did not by itself have an effect (3 microg). We conclude that intrathecal NPY acts in part via Y1 receptors to inhibit ongoing inflammatory nociception.

Topics: Animals; Behavior, Animal; Blood Pressure; Dose-Response Relationship, Drug; Inflammation; Injections, Spinal; Male; Neuropeptide Y; Pain; Perception; Rats; Rats, Sprague-Dawley; Receptors, Neuropeptide Y

The present study was carried out to investigate the effects of neuropeptide Y (NPY) in the midbrain periaqueductal grey (PAG) on the nociceptive modulation in mononeuropathic rats. NPY was microinjected into the PAG. The latency of paw withdrawal (PWL), assessed by the hot-plate (52 ) and the Randall Selitto test, was measured. Intra-PAG administration of 0.05, 0.1 and 0.2 nmol of NPY significantly increased the PWLs in a dose-dependent manner. Co-administration of 0.2 nmol of NPY(28-36) and 5.5 nmol of naloxone significantly attenuated the NPY-induced increase in PWLs. The results suggest that Y(1) receptor may mediate NPY-induced anti-nociception, and that the opioid receptors in PAG may also be involved in this process in mononeuropathic rats.

Topics: Analgesics; Animals; Male; Microinjections; Mononeuropathies; Neuropeptide Y; Nociceptors; Pain; Pain Threshold; Periaqueductal Gray; Rats; Rats, Sprague-Dawley

In the present study, the rat sciatic nerve was constricted to varying degrees using only one ligature with a very thin polyethylene sheath placed between nerve and ligature thread. Complete nerve transection was studied for comparison. With a 40-80% constriction of the nerve we observed allodynia to a similar extent as in the so-called Bennett model based on four loose ligatures. We also monitored changes in the expression of neuropeptide Y (NPY) and the NPY Y1 receptor (Y1R) in the lumbar 4-5 dorsal root ganglia (DRG) and dorsal horn and found upregulation of NPY and downregulation of the Y1R in DRG neurons after injury. These results indicate that similar peptide and receptor changes occur in this model as after axotomy and in other nerve injury models, although the immunohistochemical and behavioral changes seem to be dependent on the degree of constriction of the nerve. Thus, it seems relevant to monitor the degree of constriction when evaluating pain and other post-injury events. The possibility that some of the changes in NPY-ergic neurotransmission are related to the generation of allodynia is discussed; as well as the possibility to use this mononeuropathic model based on a single ligature nerve constriction (SLNC) as a complementary approach to other widely used pain models.

Topics: Animals; Behavior, Animal; Cell Count; Functional Laterality; Ganglia, Spinal; Immunohistochemistry; Ligation; Male; Neurons; Neuropeptide Y; Pain; Pain Measurement; Rats; Rats, Sprague-Dawley; Rats, Wistar; Receptors, G-Protein-Coupled; Receptors, Neuropeptide; Sciatic Nerve; Sciatic Neuropathy; Spinal Cord; Time Factors

Define covariates of cerebrospinal corticotropin-releasing hormone (CRH) levels in normal humans. CRHCSF was measured in 9 normal subjects as part of an intensive study of physiological responses stressors in chronic pain and fatigue states. CRHCSF was first correlated with demographic, vital sign, HPA axis, validated questionnaire domains, baseline and maximal responses to pain, exercise and other stressors. Significant factors were used for linear regression modeling.. Highly significant correlations were found despite the small number of subjects. Three models were defined: (a) CRHCSF with blood glucose and sodium (explained variance = 0.979, adjusted R2 = 0.958, p = 0.02 by 2-tailed testing); (b) CRHCSF with resting respiratory and heart rates (R2 = 0.963, adjusted R2 = 0.939, p = 0.007); and (c) CRHCSF with SF-36 Vitality and Multidimensional Fatigue Inventory Physical Fatigue domains (R2 = 0.859, adjusted R2 = 0.789, p = 0.02).. Low CRHCSF was predicted by lower glucose, respiratory and heart rates, and higher sodium and psychometric constructs of well being. Responses at peak exercise and to other acute stressors were not correlated. CRHCSF may have reflected an overall, or chronic, set-point for physiological responses, but did not predict the reserves available to respond to immediate stressors.

Topics: Adult; Blood Glucose; Confidence Intervals; Corticotropin-Releasing Hormone; Demography; Fatigue; Female; Heart Rate; Humans; Male; Models, Statistical; Neuropeptide Y; Pain; Radioimmunoassay; Regression Analysis; Respiration; Sex Factors; Sodium; Statistics as Topic; Stress, Physiological

Demyelinating diseases can be associated with painful sensory phenomena such as tactile allodynia and hyperalgesia. To study the mechanisms underlying demyelination-induced pain, we have characterized a novel model of demyelination of the sciatic or saphenous nerve. Topical lysolecithin application causes focal demyelination of afferent nerve A-fibers without axonal loss, as assessed either by electron and light microscopy or by immunohistochemical analysis of dorsal root ganglia (DRG) for a neuronal injury marker, activating transcription factor 3. Focal demyelination is accompanied by spontaneous action potentials in afferents and increased expression of neuropeptide Y and Na(v)1.3 sodium channels specifically in DRG neurons that coexpress a specific marker of myelinated afferents. In contrast, expression of tetrodotoxin-resistant, Na(v)1.8 sodium channels is specifically decreased in the same subgroup of DRG cells. Central sensitization of somatosensory processing is also induced, with increased behavioral reflex responsiveness to thermal and mechanical stimuli. These changes are reversed by intrathecal administration of an NMDA receptor antagonist or cannabinoid (CB) receptor agonist, but not by a mu-opioid receptor agonist. Recovery of behavioral reflexes occurred approximately 3 weeks after lysolecithin treatment. This is the first time that demyelination of afferent A-fibers has been shown to specifically induce neuropathic pain and indicates that axonal damage is not a prerequisite for development of the pain state. The profile of phenotypic changes in DRG is distinct from other pain models and displays a sensitivity to NMDA and CB receptor agents that may be exploitable therapeutically.

Topics: Action Potentials; Animals; Behavior, Animal; Cannabinoids; Demyelinating Diseases; Disease Models, Animal; Drug Administration Routes; Excitatory Amino Acid Antagonists; Ganglia, Spinal; Immunohistochemistry; Lysophosphatidylcholines; Mice; Mice, Inbred C57BL; Mononeuropathies; Nerve Fibers, Myelinated; Neurons, Afferent; Neuropeptide Y; Pain; Peripheral Nerves; Receptors, N-Methyl-D-Aspartate; Receptors, Opioid, mu; Reflex; Sciatic Nerve; Sodium Channels

We tested the hypothesis that increases in the spinal levels of vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) were related to the development of neuropathic pain. To this aim, we compared two groups of rats. One group showed well-developed neuropathic pain in the tail following unilateral transection of the inferior and superior caudal trunks between the S1 and S2 spinal nerves, and the other group showed poorly-developed neuropathic pain despite the same nerve injury. The increases in immunoreactivity of VIP and NPY in the S1 dorsal horn (injured segment) were not significantly different between the two groups. These results suggested that increases in the spinal levels of VIP and NPY after peripheral nerve injury were not sufficient for the development of neuropathic pain.

Topics: Animals; Cold Temperature; Hot Temperature; Immunohistochemistry; Male; Neuropeptide Y; Pain; Rats; Rats, Sprague-Dawley; Spinal Cord; Spinal Nerves; Vasoactive Intestinal Peptide

Glial cell line-derived neurotrophic factor (GDNF) is necessary for the development of sensory neurons, and appears to be critical for the survival of dorsal root ganglion (DRG) cells that bind the lectin IB4. Intrathecal infusion of GDNF has been shown to prevent and reverse the behavioral expression of experimental neuropathic pain arising from injury to spinal nerves. This effect of GDNF has been attributed to a blockade of the expression of the voltage gated, tetrodotoxin-sensitive sodium channel subtype, Na(V)1.3, in the injured DRG. Here we report that GDNF given intrathecally via osmotic-pump to nerve-injured rats (L5/L6 spinal nerve ligation) prevented the changes in a variety of neurochemical markers in the DRG upon injury. They include a loss of binding of IB4, downregulation of the purinergic receptor P2X(3), upregulation of galanin and neuropeptide Y immunoreactivity in large diameter DRG cells, and expression of the transcription factor ATF3. GDNF infusion concomitantly prevented the development of spinal nerve ligation-induced tactile hypersensitivity and thermal hyperalgesia. These observations suggest that high dose, exogenous GDNF has a broad neuroprotective role in injured primary afferent. The receptor(s) that mediates these effects of GDNF is not known. GDNF's ability to block neuropathic pain states is not likely to be specific to Na(V)1.3 expression.

Topics: Activating Transcription Factors; Animals; Blood Proteins; Disease Models, Animal; Functional Laterality; Galanin; Ganglia, Spinal; Glial Cell Line-Derived Neurotrophic Factor; Humans; Immunohistochemistry; Lectins; Male; Nerve Growth Factors; Neurons; Neuropeptide Y; Pain; Pain Threshold; Rats; Rats, Sprague-Dawley; Receptors, Purinergic P2; Receptors, Purinergic P2X3; Time Factors; Transcription Factors

Three subtypes of neurotensin receptor have been described, two members of the heptahelical transmembrane domain G protein-coupled receptor superfamily NT-1R and NT-2R, and NT-3R unrelated to this family. We have generated NT-1R deficient (NT-1R(-/-)) mice. NT-1R(-/-) mice were born at the expected Mendelian frequency without obvious abnormalities and they were fertile. The NT-induced analgesia on the writhing induced by phenyl-p-benzoquinone administration remained at wild-type levels in the NT-1R(-/-) mice demonstrating that the NT-1R is not implicated in the analgesic effect of NT in this test. The NT-1R(-/-) mice were hyperthermic; their body temperature was not affected by intracerebroventricular (i.c.v.) administration of NT, contrasting with the hypothermia induced in NT-1R(+/+) mice. NT-1R(-/-) mice showed a small significant increase in body weight compared to the NT-1R(+/+) congeners as early as 10 weeks after birth, correlated with a higher food intake. NT-1R(-/-) mice showed similar spontaneous locomotion to the control littermates, but did not respond to i.c.v. NT-induced hypolocomotion. I.c.v. injection of NT inhibited feeding in fasted wild-type mice, but had no effect on feeding of the NT-1R(-/-) mice. I.c.v. administration of the orexigenic neuropeptide Y (NPY) stimulated feeding to the same extent in both wild-type and NT-1R(-/-) mice. This analysis of NT-1R-deficient mice shows that the NT-1R does not play a role in NT-induced analgesia, but that it is clearly implicated in thermal and feeding regulation, weight control, and NT-induced hypolocomotion.

Topics: Analgesics; Animals; Body Temperature Regulation; Feeding Behavior; Female; Gene Deletion; Hypothermia; Injections, Intraventricular; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Motor Activity; Neuropeptide Y; Neurotensin; Pain; Receptors, Neurotensin

The present experiments investigated the behavioral and immunocytchemical (ICC) effects of applying complete Freund's adjuvant (CFA) to the orbital portion of the infraorbital nerve (IOn). Two control groups, the first had saline applied to the IOn and the second underwent sham operation, were included in the study. In the CFA group, significant hyper-responsiveness to von Frey (analysis of variance <0.05) and to pinprick stimulation (Kruskal Wallis <0.05) in the vibrissal pad was observed on the fourth and the fifth days post-operative (dpo). This was accompanied by a reduced bite force and altered bite patterns of similar duration. Histology of the IOn in CFA rats revealed immune cell infiltration and edema around and in the nerve trunk with only mild axonal damage confirmed by neuropeptide Y immunoreactivity in trigeminal ganglion. Histological areas of inconsistent and mild inflammation were observed in the saline group that were accompanied by similarly attenuated behavioral and ICC changes. This model of inflammation-induced neuropathic pain is highly applicable to the study of neuroinflammatory orofacial pain.

Topics: Animals; Facial Pain; Freund's Adjuvant; Hyperalgesia; Inflammation; Inflammation Mediators; Male; Neuropeptide Y; Orbit; Pain; Pain Threshold; Rats; Rats, Sprague-Dawley; Trigeminal Nerve

Histamine H1 receptor mRNA-expressing sensory neurons in guinea pigs are unmyelinated and are not immunoreactive to substance P and calcitonin gene-related peptide (CGRP) [Mol. Brain Res. 66 (1999) 24], which are implicated in the nociceptive transmission of the primary sensory system. In this study, we examined whether these H1 receptor mRNA-expressing neurons are sensitive to capsaicin by using in situ hybridization histochemistry. Of lumbar dorsal root ganglion (DRG) neurons in control animals, 17% were positive for CGRP. In guinea pigs neonatally treated with capsaicin (50 mg/kg), few CGRP-immunoreactive neurons were seen in the DRGs. However, the percentages of H1 receptor mRNA-expressing neurons (15-20%) and the intensity of the mRNA signals in these neurons were not affected by neonatal capsaicin treatment. We also revealed the presence of both capsaicin-sensitive and insensitive neuropeptide Y (NPY)-immunoreactive neurons in the DRGs. These neurons were exclusively small. H1 receptor mRNA was expressed in NPY-immunoreactive neurons in naive guinea pig DRGs. These results suggest that H1 receptor mRNA is expressed in capsaicin-insensitive DRG neurons with NPY-immunoreactivity in guinea pigs.

Topics: Animals; Calcitonin Gene-Related Peptide; Capsaicin; Ganglia, Spinal; Guinea Pigs; Histamine; Immunohistochemistry; Male; Nerve Fibers; Neurons, Afferent; Neuropeptide Y; Nociceptors; Pain; Posterior Horn Cells; Receptors, Histamine H1; RNA, Messenger

Expression of C-terminal flanking peptide of neuropeptide Y (CPON) in DRG and cell proliferation (incorporation of BrdU) in sciatic nerve of rats following chronic nerve compression (silicone tubes with different internal diameters) was studied by immunocytochemistry. An increased number of CPON-positive neurons and cells incorporating BrdU was induced on the compressed side, most pronounced when a tight tube was used, while no cells expressed CPON or BrdU in intact nerves. The increase was transient and declined with time. Nerve compression induces transient cell proliferation in the nerve and expression of CPON in nerve cell bodies, but this is of a lesser magnitude than those following nerve transection.

Topics: Animals; Axons; Bromodeoxyuridine; Cell Count; Cell Division; Chronic Disease; Denervation; Disease Models, Animal; Female; Ganglia, Spinal; Immunohistochemistry; Nerve Compression Syndromes; Nerve Degeneration; Neurons, Afferent; Neuropeptide Y; Pain; Peptide Fragments; Rats; Rats, Wistar; Schwann Cells; Sciatic Nerve; Up-Regulation

Hindpaw withdrawal latency (HWL) to thermal and mechanical stimulation increased dose-dependently after intra-periaqueductal grey (PAG) injection of neuropeptide Y (NPY). Furthermore, the NPY-induced increases in HWLs were attenuated by intra-PAG injection of the Y1 receptor antagonist NPY28-36. The results demonstrated that NPY plays an important role in antinociception in PAG, in which Y1 receptor is involved in.

Topics: Analgesia; Animals; Male; Neuropeptide Y; Nociceptors; Pain; Pain Measurement; Periaqueductal Gray; Rats; Rats, Sprague-Dawley

We examined the expression of three neuropeptides that have been implicated in nociceptive transmission, and the sympathetic nerve fiber marker tyrosine hydroxylase, in 11 painful human Morton's neuromas, using immunohistochemistry. Antibodies against the neural markers RT97 and PGP 9.5 were used to map the general nerve fiber organization of the neuromas. Four specimens of normal human peripheral nerves were used as controls. Substance P, calcitonin gene-related peptide, and neuropeptide Y immunoreactivities were more pronounced in neuroma tissue than in control nerves. Neuropeptide immunofluorescence was seen both in larger nerve fiber trunks and in masses of disorganized axon profiles dispersed in loose connective tissue. Tyrosine hydroxylase immunoreactivity was present at varying levels of expression in neuroma nerve fiber trunks, in connective tissue nerve fiber bundles, and around some blood vessels. Our findings suggest that neuropeptides are involved in the response to injury in Morton's neuromas and that they could play a role in initiation or modulation of pain. In addition, pain from Morton's neuromas could be influenced by sympathetic nerve fibers.

Topics: Adult; Aged; Antibodies; Calcitonin Gene-Related Peptide; Cell Communication; Female; Foot Diseases; Humans; Male; Middle Aged; Nerve Fibers; Neuroma; Neurons, Afferent; Neuropeptide Y; Neuropeptides; Pain; Substance P; Sympathetic Nervous System; Thiolester Hydrolases; Tyrosine 3-Monooxygenase; Ubiquitin Thiolesterase

The nociceptive response latencies increased significantly after intra-nucleus raphe magnus administration of 0.1 or 0.4 nmol of neuropeptide Y, but not 0.04 nmol, in rats. The neuropeptide Y-induced increases in hindpaw withdrawal latency were reversed by following injection of 0.42 nmol of the Y1 antagonist, NPY(28-36). The results indicate that NPY plays an antinociceptive role in nucleus raphe magnus in rats, which is mediated by the Y1 receptor. Furthermore, the neuropeptide Y-induced increases in hindpaw withdrawal latency were attenuated by following intra-nucleus raphe magnus injection of 6 nmol of the opioid antagonist naloxone, indicating that there is an interaction between NPY and opioids in nucleus raphe magnus.

Topics: Animals; Hot Temperature; Male; Microinjections; Naloxone; Neuropeptide Y; Pain; Peptide Fragments; Raphe Nuclei; Rats; Rats, Sprague-Dawley; Reaction Time; Receptors, Neuropeptide Y

Recent animal models of experimental nerve injury have proven useful in evaluating potential sympathetic involvement in neuropathic pain syndromes. We have employed a widely adopted unilateral L5/L6 spinal nerve ligation model to compare the development of mechanical allodynia with neurochemical changes both at the site of peripheral nerve injury and in the dorsal root ganglia (DRG). We have focused on the expression of neuropeptide Y (NPY), a well-studied regulatory peptide and phenotypic marker of sympathetic neurons, and functionally related Y-receptor binding sites following nerve injury. In sympathetic neurons, NPY is colocalized and coreleased with norepinephrine (NE) at peripheral sites of action. Furthermore, NPY gene expression is induced within the population of medium- and large-diameter DRG neurons of the A beta-fiber class after experimental nerve injury. We therefore hypothesized that concurrent alterations in NPY and NE expression by sympathetic and sensory neurons may be a contributing factor to sympathetically-maintained neuropathic conditions. Animals with unilateral L5/L6 spinal nerve ligation developed mechanical allodynia of the hind paw ipsilateral to the site of injury that persisted until sacrifice at postoperative day 10. A significant induction of preproneuropeptide Y-encoding (PPNPY) mRNA, as detected by in situ hybridization histochemistry (ISHH), occurred in populations of medium- and large-diameter DRG neurons ipsilateral to the site of injury. Immunohistochemical analysis indicated a marked decline in the number of labeled sympathetic axons positive for tyrosine hydroxylase-like and NPY-like immunoreactivities (TH-LI and NPY-LI, respectively) proximal to the site of nerve injury and almost complete elimination of immunopositive fibers distal to the site of ligation. Whereas, the extent of colocalization of NPY-LI to TH-LI-positive sympathetic axons in unaffected L4 or L5 nerve segments exceeded 80%, this figure declined to approximately 50% in regenerating axons of ligated spinal nerve L5. The portion of NPY-LI that was not colocalized to sympathetic TH-LI-positive fibers was most likely contributed by regenerating sensory axons, consistent with marked de novo synthesis of NPY by DRG neurons. In end bulb axon terminals, i.e. morphological profiles characteristic of neuromas, NPY-LI-positive elements that were not colocalized to TH-LI-positive sympathetic elements appeared to be spatially segregated from those of sympathetic

Topics: Animals; Autoradiography; Axons; Ganglia, Spinal; Immunohistochemistry; In Situ Hybridization; Ligation; Male; Neurons; Neuropeptide Y; Pain; Physical Stimulation; Protein Precursors; Rats; Rats, Sprague-Dawley; Receptors, Neuropeptide Y; RNA, Messenger; Spinal Nerves; Tyrosine 3-Monooxygenase

We have examined the effects of intrathecal (i.t.) administration of [Leu31,Pro34]-neuropeptide Y (NPY) or NPY-(13-36), selective agonists of NPY Y1 or Y2 receptors, respectively, on the excitability of the flexor reflex in normal rats and after unilateral transection of the sciatic nerve. In rats with intact and sectioned sciatic nerves, i.t. [Leu31,Pro34]-NPY induced a similar biphasic effect on the flexor reflex with facilitation at low doses and facilitation followed by depression at high doses. In contrast, i.t. NPY-(13-36) only facilitated the flexor reflex in normal rats, and at high dose it caused ongoing discharges in the electromyogram. NPY-(13-36) caused dose-dependent depression of the flexor reflex in rats after sciatic nerve transection, in addition to its facilitatory effect. Topical application of [Leu31,Pro34]-NPY or NPY-(13-36) caused a moderate and brief reduction in spinal cord blood flow. No difference was noted between the vasoconstrictive effect of [Leu31,Pro34]-NPY and NPY-(13-36). It is suggested that activation of Y1 receptors may be primarily responsible for the reflex depressive effect of i.t. neuropeptide Y in rats with intact sciatic nerves, whereas both Y1 and Y2 receptors may be involved in mediating the depressive effect of NPY after axotomy.

Topics: Animals; Axotomy; Dose-Response Relationship, Drug; Female; Injections, Spinal; Neuropeptide Y; Nociceptors; Pain; Peptide Fragments; Peroneal Nerve; Rats; Rats, Sprague-Dawley; Receptors, Neuropeptide Y; Reflex; Spinal Cord; Tibial Nerve

Topics: Anesthesiology; Humans; Hyperalgesia; Nerve Growth Factors; Neuropeptide Y; Neurotrophin 3; Nociceptors; Pain; Pain Management; Receptors, N-Methyl-D-Aspartate

The expression of galanin and neuropeptide Y in rat lumbar 5 (L5) dorsal root ganglia and dorsal horn (L4-5) was studied after four types of peripheral nerve injury using immunohistochemistry and in situ hybridization. The possible correlation between these two peptides and tactile allodynia-like behaviour was analysed as well. The models employed were the Gazelius (photochemical lesion) and Seltzer and Bennett (constriction lesions) models, as well as complete sciatic nerve transection (axotomy). Two weeks after surgery, the Gazelius model rats more frequently displayed a greater tactile allodynia than the rats from the Seltzer and Bennett models. Tactile allodynia was not observed in any of the axotomized rats. A marked increase in the number of galanin-immunoreactive and galanin messenger RNA-positive neuron profiles was observed in ipsilateral dorsal root ganglia in all types of models. The increase in allodynic rats (Gazelius, Seltzer and Bennett models) was less pronounced than that after axotomy. In addition, in the Bennett model the number of galanin-immunoreactive neurons was significantly lower in allodynic rats as compared to non-allodynic rats, and the same tendency, but less obvious was found in the Seltzer model. Furthermore, an increase in galanin-immunoreactive fibres was found in the superficial laminae of the ipsilateral dorsal horn in all lesion models, especially in lamina II. A dramatic increase in the number of neuropeptide Y and neuropeptide Y messenger RNA-positive neuron profiles was also found in the ipsilateral dorsal root ganglia in all models, but no significant difference was found in peptide levels between allodynic and non-allodynic rats in any of the models. The present results suggest that the levels of endogenous galanin may play a role in whether or not allodynia develops in the Bennett model.

Topics: Animals; Axotomy; Behavior, Animal; Cell Size; DNA Probes; Galanin; Ganglia, Spinal; Immunohistochemistry; In Situ Hybridization; Male; Neurons; Neuropeptide Y; Pain; Peripheral Nervous System Diseases; Physical Stimulation; Rats; Rats, Sprague-Dawley; RNA, Messenger; Spinal Cord

Neuropathic pain resulting from peripheral nerve injury can often be relieved by administration of alpha-adrenergic receptor antagonists. Tonic activation of alpha-adrenergic receptors may therefore facilitate the hyperalgesia and allodynia associated with neuropathic pain. It is currently unclear whether alpha2A- or alpha2c-adrenergic receptor subtypes are involved in the pro-nociceptive actions of alpha-adrenergic receptors under neuropathic conditions. We therefore investigated the effects of peripheral nerve injury on the expression of these subtypes in rat spinal cord using immunohistochemical techniques. In addition, neuropeptide Y immunoreactivity was examined as an internal control because it has previously been shown to be up-regulated following nerve injury. We observed a decrease in alpha2A-adrenergic receptor immunoreactivity in the spinal cord ipsilateral to three models of neuropathic pain: complete sciatic nerve transection, chronic constriction injury of the sciatic nerve and L5/L6 spinal nerve ligation. The extent of this down-regulation was significantly correlated with the magnitude of injury-induced changes in mechanical sensitivity. In contrast, alpha2c-adrenergic receptor immunoreactivity was only increased in the spinal nerve ligation model; these increases did not correlate with changes in mechanical sensitivity. Neuropeptide Y immunoreactivity was up-regulated in all models examined. Increased expression of neuropeptide Y correlated with changes in mechanical sensitivity. The decrease in alpha2A-adrenergic receptor immunoreactivity and the lack of consistent changes in alpha2C-adrenergic receptor immunoreactivity suggest that neither of these receptor subtypes is likely to be responsible for the abnormal adrenergic sensitivity observed following nerve injury. On the contrary, the decrease in alpha2A-adrenergic receptor immunoreactivity following nerve injury may result in an attenuation of the influence of descending inhibitory noradrenergic input into the spinal cord resulting in increased excitatory transmitter release following peripheral stimuli.

Topics: Animals; Chronic Disease; Hyperalgesia; Immunohistochemistry; Ligation; Male; Nerve Compression Syndromes; Neuropeptide Y; Pain; Physical Stimulation; Rats; Rats, Sprague-Dawley; Receptors, Adrenergic, alpha-2; Sciatic Nerve; Spinal Cord; Spinal Nerves

Neuropeptide Y (NPY) increases the excitability of 'small', nociceptive, dorsal root ganglion (DRG) neurons. This effect, which may contribute to the etiology of 'neuropathic' pain, has been attributed to attenuation of Ca2+-sensitive K+ conductance(s) (gK,Ca) following suppression of Ca2+ entry via N-type Ca2+ channels. A problem arises with this conclusion because rat DRG neurons normally contain high intracellular Cl- and some of them express a Ca2+-dependent Cl- conductance (gCl,Ca). In this study, we find that in rat DRG neurons increasing intracellular Cl- does not attenuate the effect of 1 microM NPY because gCl,Ca is not found in 'small' DRG cells and the peptide failed to affect the gCl,Ca found in 'large' cells. Thus, the presence of gCl,Ca in a subpopulation of 'large' DRG neurons does not alter the conclusion that excitatory effects of NPY result from attenuation of gK,Ca.

Topics: Action Potentials; Animals; Axotomy; Calcium; Cell Size; Cells, Cultured; Chloride Channels; Chlorides; Electrophysiology; Ganglia, Spinal; Ion Channel Gating; Nerve Compression Syndromes; Neurons, Afferent; Neuropeptide Y; Nodose Ganglion; Pain; Rats; Rats, Sprague-Dawley; Sympathetic Nervous System; Trigeminal Ganglion

The central nervous system regulates gut functions via complex interactions between the enteric nervous and immune systems. The aim of this study was to investigate whether mast cell mediators are released into the human jejunal lumen during stress.. A closed-segment perfusion technique was used to investigate jejunal release of tryptase, histamine, prostaglandin D2, and water flux in response to the cold pressor test in 8 healthy subjects and 9 patients with food allergy. In 6 food-allergic patients, jejunal biochemical responses to cold pain stress were compared with those induced by food intraluminal challenge.. Cold pain stress elevated heart rate and blood pressure and increased luminal release of mast cell mediators and jejunal water secretion in both groups. Stress-induced release of tryptase and histamine, but not of prostaglandin D2 and water flux, was greater in food-allergic patients than in healthy volunteers. In food-allergic patients, jejunal biochemical responses induced by cold pain stress were similar to those induced by antigen challenge.. These results show the ability of the central nervous system to modulate intestinal mast cell activity and suggest that mast cells have a role in stress-related gut dysfunction.

Topics: Adult; Body Water; Chymases; Cold Temperature; Female; Histamine Release; Humans; Jejunum; Male; Mast Cells; Neuropeptide Y; Ovalbumin; Pain; Serine Endopeptidases; Stress, Physiological; Tryptases

Neuropeptide Y (NPY) and endogenous opioids (EOPs) such as methionine-enkephalin (Met-enk) regulate similar physiological responses, but it is not known whether nociceptive and immune responses also show analogy after intracerebroventricular (i.c.v.) application. Dose-response studies show that Met-enk stimulates the blood granulocyte and splenic natural killer (NK) cell function of Lewis rats at a low dose (10(2) ng/kg, i.c.v.), whereas a high dose (10(5) ng/kg) causes suppression of innate immune functions associated with analgesia in the hot-plate test. At 15 min, 1 h and 24 h after i.c.v. application, both Met-enk (10(2) ng/kg) and NPY (1 ng/kg) produced similar effects: An initial suppression of innate immune function was followed by a long lasting stimulatory action on cell functions and serum interleukin-6 (sIL-6) levels. Thus, central NPY application resembles Met-enk-induced immunostimulation at doses not affecting nociception, suggesting an involvement of both peptides in shaping stress-induced immunomodulation of the non-analgetic form, possibly via activation of a common immunomodulatory effector mechanism.

Topics: Adjuvants, Immunologic; Analgesics; Animals; Dose-Response Relationship, Drug; Enkephalin, Methionine; Injections, Intraventricular; Male; Neuropeptide Y; Pain; Rats; Rats, Inbred Lew

The levels of dorsal root ganglia and paravertebral sympathetic ganglia innervating the lumbar facet joint were investigated in rats using the retrograde transport method. The pathways and functions of the nerve fibers supplying the lumbar facet joint were determined immunohistochemically.. To study lumbar facet pain in relation to its innervation.. The lumbar facet joints have been reported to be innervated segmentally. Little is known, however, about the origins and functions of the nerve fibers.. Cholera toxin B subunit, a neural tracer, was placed in the L5-L6 facet joint, and the bilateral dorsal root ganglia and paravertebral sympathetic ganglia were examined immunohistochemically. The serial sections of lumbar vertebrae of newborn rats and the sections of the facet joint capsules, dorsal root ganglia, and paravertebral sympathetic ganglia of adult rats were investigated immunohistochemically. The pathways of the nerve fibers supplying the facet joint were reconstituted.. Labeled neurons existed in ipsilateral dorsal root ganglia from L1 to L5 and in paravertebral sympathetic ganglia from T12 to L6. The dorsal ramus of the spinal nerve and rami communicantes were connected to each other by calcitonin gene-related peptide immunoreactive fibers and dopamine beta-hydroxylase immunoreactive fibers.. The L5-L6 facet joint was innervated by ipsilateral dorsal root ganglia and paravertebral sympathetic ganglia, segmentally and nonsegmentally. Some of the sensory fibers from the facet joint may pass through the paravertebral sympathetic trunk, reaching L1 and/or L2 dorsal root ganglia. Inguinal and/or anterior thigh pain with lower lumbar facet joint lesions may be explained as referred pain.

Topics: Animals; Antibody Specificity; Calcitonin Gene-Related Peptide; Cell Count; Cholera Toxin; Dopamine beta-Hydroxylase; Ganglia, Spinal; Ganglia, Sympathetic; Immunohistochemistry; Lumbar Vertebrae; Male; Nerve Fibers; Neurons, Afferent; Neuropeptide Y; Pain; Rats; Rats, Sprague-Dawley; Substance P; Vasoactive Intestinal Peptide

In normal animals, spinal administration of neuropeptide Y induces analgesia to thermal stimuli, but has no effect on mechanical thresholds. Recent anatomical studies, however, have shown that following nerve injury there is an altered expression of neuropeptide Y and its receptors. The aim of this behavioural study, therefore, is to examine the effect of intrathecal administration of neuropeptide Y, its agonists and an antagonist on mechanical nociceptive thresholds in rats with partial injury to the sciatic nerve. Test agents were administered for 14 days via osmotic pumps (0.5 microliter/day) attached to intrathecal catheters and the nociceptive flexion reflex was quantified using an Ugo Basile Analgesymeter. Partial injury to the sciatic nerve, in animals treated intrathecally with saline, induces a significant decrease in mechanical threshold as compared to the sham operated, contralateral paw. The nerve injury-induced hyperalgesia is exacerbated by 2 microM neuropeptide Y and by 2 microM [Leu31,Pro34]-neuropeptide Y, a Y1 receptor agonist. The Y2 receptor agonist, N-acetyl-[Leu28,Leu31]-neuropeptide Y24-36 (2 microM), had no effect on the nerve injury-induced hyperalgesia. The putative neuropeptide Y antagonist, alpha-trinositol (10 microM), significantly attenuated the nerve injury-induced hyperalgesia. This study suggests that neuropeptide Y may contribute to nerve injury-induced mechanical hyperalgesia via the Y1 receptor and provides further insight into the possible mechanisms underlying nerve injury-induced hyperalgesia to mechanical stimuli.

Topics: Analysis of Variance; Animals; Anti-Inflammatory Agents, Non-Steroidal; Functional Laterality; Hyperalgesia; Injections, Spinal; Inositol Phosphates; Neuropeptide Y; Pain; Rats; Rats, Wistar; Receptors, Neuropeptide Y; Sciatic Nerve; Sensory Thresholds; Spinal Cord

The release of immunoreactive (ir-) neuropeptide Y (NYP) was studied in the anaesthetized rat and cat by means of microprobes bearing immobilized antibodies to the C terminus of NPY. An extensive basal release of ir-NYP was detected throughout the dorsal and upper ventral horn of the rat. This spontaneous release was not significantly altered by sectioning the spinal cord at the thoraco-lumbar junction nor by electrical stimulation of peripheral nerves. Since NPY is virtually absent in primary afferents it is probable that spontaneous release within the spinal cord comes from active NPY-containing intrinsic spinal neurones. In the spinal cat spontaneous release of ir-NPY was detected in the mid-dorsal horn and this was unaltered by peripheral noxious thermal or noxious mechanical stimuli. As in the rat, release from intrinsic spinal neurones is most probable. The extensive spontaneous release of ir-NPY in both species suggests a widespread role in spinal cord function.

Topics: Anesthesia, General; Animals; Cats; Electric Stimulation; Male; Neuropeptide Y; Pain; Peripheral Nerves; Physical Stimulation; Rats; Rats, Wistar; Spinal Cord

In normal animals, peripheral nerve injury produces a persistent, neuropathic pain state in which pain is exaggerated and can be produced by nonpainful stimuli. Here, mice that lack protein kinase C gamma (PKCgamma) displayed normal responses to acute pain stimuli, but they almost completely failed to develop a neuropathic pain syndrome after partial sciatic nerve section, and the neurochemical changes that occurred in the spinal cord after nerve injury were blunted. Also, PKCgamma was shown to be restricted to a small subset of dorsal horn neurons, thus identifying a potential biochemical target for the prevention and therapy of persistent pain.

Topics: Animals; Ganglia, Spinal; Gene Deletion; Hyperalgesia; Inflammation; Interneurons; Isoenzymes; Ligation; Mice; Mice, Knockout; Neuropeptide Y; Pain; Pain Management; Pain Threshold; Protein Kinase C; Receptors, Neurokinin-1; Sciatic Nerve; Signal Transduction; Spinal Cord; Substance P

The present study shows further evidence about the implication of neuropeptide Y (NPY) in nociception. The effect of NPY (1-36), when intracerebroventricularly administered, has been studied using two physical models of acute pain (hot plate test and electrical tail stimulation) and the formalin test. The animal response to these three pain models has been shown to be integrated at different levels in the CNS. A decrease in pain threshold was exhibited in both the hot plate test (10, 30, 60, 120 and 480 pmol of NPY i.c.v.) and the electrical tail simulation test (10, 30 and 60 pmol of NPY i.c.v.), while in the formalin test (10, 30, 60 and 120 pmol of NPY icv) the licking response decreased in phase I but not in phase 2. In these three tests NPY showed hyperalgesic or analgesic effects when administered at low doses, while at high doses it failed to induce any effect. Results show that the effect of NPY on nociception is clearly test-dependent and is only observed at low doses.

Topics: Analgesia; Animals; Electric Stimulation; Hyperalgesia; Injections, Intraventricular; Male; Mice; Neuropeptide Y; Pain

The contribution of the nervous system to the pathophysiology of rheumatoid arthritis has been proposed to be mediated by certain neuropeptides. Neuropeptide Y, calcitonin gene-related peptide, substance P, and neurokinin A are considered modulators of inflammatory joint disease. Parameters of pain, as well as occlusal signs of tissue destruction from the arthritic TMJ and the corresponding neuropeptide concentrations in TMJ synovial fluid, were investigated in patients with various inflammatory joint diseases. The patients with rheumatoid arthritis were also examined in a separate diagnostic group. Visual analog scale, palpatory tenderness, maximal voluntary mouth opening, and anterior open bite were correlated to neuropeptide-like immunoreactivities of the above four neuropeptides. It was found that high concentrations of calcitonin gene-related peptide and neuropeptide Y in TMJ fluid are associated with pain, impairment of mandibular mobility, and occlusal signs of TMJ destruction in patients with rheumatoid arthritis. The results indicated neuropeptide involvement in rheumatoid arthritis, proposing a potentiation of the symptoms and signs by the inflammatory action of calcitonin gene-related peptide and neuropeptide Y.

Topics: Adult; Arthritis; Arthritis, Rheumatoid; Calcitonin Gene-Related Peptide; Female; Humans; Inflammation Mediators; Male; Middle Aged; Neuroimmunomodulation; Neurokinin A; Neuropeptide Y; Neuropeptides; Pain; Pain Measurement; Range of Motion, Articular; Statistics, Nonparametric; Substance P; Synovial Fluid; Temporomandibular Joint Disorders

We sought to identify characteristics of peptidergic innervation that altered in patients with chronic pancreatitis. Pancreatic tissue removed from patients with chronic pancreatitis was analyzed by immunohistochemistry using antisera against neuropeptide Y, tyrosine hydroxylase, vasoactive intestinal polypeptide, peptide histidine isoleucine, calcitonin gene-related peptide, and substance P, respectively. In accordance with recent findings, the number and diameter of intralobular and interlobular nerve bundles were found to be increased as compared with control pancreas from organ donors. The striking change in the peptidergic innervation pattern in chronic pancreatitis concerned these altered nerves. It consisted of an intensification of the immunostaining for calcitonin gene-related peptide and substance P in numerous fibers contained in these nerves. Adjacent sections showed that immunoreactive substance P and immunoreactive calcitonin gene-related peptide coexisted in these fibers. Because both of these peptides are generally regarded as pain transmitter candidates, our findings provide further evidence that changes in pancreatic nerves themselves might be responsible for the long-lasting pain syndrome in chronic pancreatitis.

Topics: Adult; Calcitonin Gene-Related Peptide; Chronic Disease; Female; Humans; Male; Nerve Fibers; Neuropeptide Y; Neuropeptides; Pain; Pancreas; Pancreatitis; Peptide PHI; Substance P; Tyrosine 3-Monooxygenase; Vasoactive Intestinal Peptide

Changes in neuropeptide Y-like immunoreactivity (NPYir) in the rat L4 and L5 spinal cord and dorsal root ganglia (DRG) were examined after different sciatic nerve injuries (transection, loose ligation, and crush) and a localized, painful inflammation of the hind paw. Inflammation had no effect on NPYir. All the nerve injuries produced comparable increases in NPYir in ipsilateral laminae III-V axons and varicosities, and induction of NPYir in many DRG cells. Most NPYir DRG cells were medium to large (mean diameters: 40-45 microns); less than 2% of the cells had diameters of 25 microns or less. We conclude that the nerve injury-evoked increase in NPYir occurs mostly in the somata and intraspinal arbors of low-threshold mechanoreceptors; very few, if any, C-fiber afferents are involved. Nerve injury, rather than a painful condition, appears to be the stimulus for the induction of NPYir synthesis.

Topics: Animals; Ganglia, Spinal; Inflammation; Male; Neurons, Afferent; Neuropeptide Y; Pain; Rats; Rats, Sprague-Dawley; Sciatic Nerve; Spinal Cord

Neuropeptide Y (NPY) is a 36-amino-acid, C-terminal amidated peptide that is found in bulbospinal pathways and can inhibit the release of the primary afferent C-fiber neurotransmitter, substance P. Based on these observations, the present studies examined the possible antinociceptive effects of this peptide and several NPY fragments after intrathecal administration in rats prepared with chronic intrathecal catheters. In the 52 degrees C hot plate test, NPY produced a dose-dependent elevation in the nociceptive threshold with a median effective dose of 1.1 nmol. The ordering of fragments' activity was: NPY greater than NPY16-36 greater than or equal to NPY19-36 greater than or equal to NPY14-36 greater than or equal to NPY18-36 much greater than NPY1-36-OH = NPY18-36-OH = 0. In the paw pressure test, NPY was not active, even at the highest doses examined (median effective dose greater than 20 nmol), whereas the C-terminal fragments retained their potency and produced significant increases in the pressure required to evoke escape (NPY18-36: median effective dose = 18.7 nmol). The rank ordering of activity in the paw pressure test was: NPY19-36 greater than or equal to NPY14-36 greater than or equal to NPY18-36 greater than or equal to NPY16-36 much greater than NPY = NPY18-36-OH = 0. Peptide YY, human pancreatic polypeptide and avian pancreatic polypeptide behave similarly to NPY.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adrenergic alpha-Antagonists; Animals; Behavior, Animal; Clonidine; Dioxanes; Dose-Response Relationship, Drug; Idazoxan; Injections, Spinal; Male; Motor Activity; Naloxone; Neuropeptide Y; Pain; Parasympathomimetics; Peptide Fragments; Rats; Rats, Inbred Strains; Structure-Activity Relationship

Neuropeptide Y (NPY) is colocalised with noradrenaline in post-ganglionic sympathetic neurons. In order to examine the possibility that activation of the sympathetic nervous system might cause release of NPY into the plasma NPY levels were measured in 16 patients undergoing exercise tests for investigation of chest pain. Plasma NPY concentrations rose in 14 out of the 16 patients, and the mean level of plasma NPY increased from 335 (s.e.m. = 37) to 455 (s.e.m. = 41) pg/ml. Plasma noradrenaline and adrenaline levels increased four- and two-fold respectively. The increase in NPY correlated with the increase in noradrenaline, suggesting that NPY may be released with noradrenaline when sympathetic noradrenergic nerves are activated.

Topics: Adult; Exercise Test; Female; Humans; Male; Middle Aged; Nerve Tissue Proteins; Neuropeptide Y; Pain; Physical Exertion; Thorax