neuropeptide-y and Peripheral-Nerve-Injuries

The dorsal root ganglion is widely recognized as a potential target to treat chronic pain. A fundamental understanding of quantitative molecular and genomic changes during the late phase of pain is therefore indispensable. The authors performed a systematic literature review on injury-induced pain in rodent dorsal root ganglions at minimally 3 weeks after injury. So far, slightly more than 300 molecules were quantified on the protein or messenger RNA level, of which about 60 were in more than one study. Only nine individual sequencing studies were performed in which the most up- or downregulated genes varied due to heterogeneity in study design. Neuropeptide Y and galanin were found to be consistently upregulated on both the gene and protein levels. The current knowledge regarding molecular changes in the dorsal root ganglion during the late phase of pain is limited. General conclusions are difficult to draw, making it hard to select specific molecules as a focus for treatment.

Topics: Animals; Galanin; Ganglia, Spinal; Mice; Neuropeptide Y; Pain Measurement; Peripheral Nerve Injuries; Rats; Rodentia; Sequence Analysis, RNA

The present study was designed to investigate involvement of angiotensin (Ang) II type 2 receptors (AT2R) in restoration of perivascular nerve innervation injured by topical phenol treatment. Male Wistar rats underwent in vivo topical application of 10% phenol around the superior mesenteric artery to induce nerve injure. Phenol treatment markedly reduced densities of both calcitonin gene-related peptide (CGRP)-like immunoreactivity (LI)- and neuropeptide Y (NPY)-LI-containing fibers. NGF restored densities of both nerve fibers to the Sham control level. Coadministration of Ang II and losartan (AT1R antagonist) significantly increased the density of CGRP-LI-fibers but not NPY-LI-fibers compared with saline control. The increase of the density of CGRP-LI-fibers by coadministration of Ang II and losartan was suppressed by adding PD123319 (AT2R antagonist). Furthermore, NGF-induced CGRP-LI nerve regeneration was inhibited by PD123319 treatment. NGF-induced increase of AT2R mRNA level was significantly suppressed by AT1R antagonist treatment in phenol treated rats dorsal root ganglia. These results suggest that selective stimulation of AT2R by Ang II facilitates reinnervation of mesenteric perivascular CGRP-containing nerves injured by topical phenol application in the rat.

Topics: Angiotensin II; Angiotensin II Type 1 Receptor Blockers; Animals; Calcitonin Gene-Related Peptide; Humans; Imidazoles; Losartan; Male; Mesenteric Arteries; Nerve Growth Factor; Nerve Regeneration; Neuropeptide Y; Peripheral Nerve Injuries; Peripheral Nerves; Phenol; Pyridines; Rats; Rats, Wistar; Receptor, Angiotensin, Type 2; Stimulation, Chemical

Neuropeptide Y targets the Y1 receptor (Y1) in the spinal dorsal horn (DH) to produce endogenous and exogenous analgesia. DH interneurons that express Y1 (Y1-INs; encoded by Npy1r) are necessary and sufficient for neuropathic hypersensitivity after peripheral nerve injury. However, as Y1-INs are heterogenous in composition in terms of morphology, neurophysiological characteristics, and gene expression, we hypothesized that a more precisely defined subpopulation mediates neuropathic hypersensitivity. Using fluorescence in situ hybridization, we found that Y1-INs segregate into 3 largely nonoverlapping subpopulations defined by the coexpression of Npy1r with gastrin-releasing peptide (Grp/Npy1r), neuropeptide FF (Npff/Npy1r), and cholecystokinin (Cck/Npy1r) in the superficial DH of mice, nonhuman primates, and humans. Next, we analyzed the functional significance of Grp/Npy1r, Npff/Npy1r, and Cck/Npy1r INs to neuropathic pain using a mouse model of peripheral nerve injury. We found that chemogenetic inhibition of Npff/Npy1r-INs did not change the behavioral signs of neuropathic pain. Further, inhibition of Y1-INs with an intrathecal Y1 agonist, [Leu31, Pro34]-NPY, reduced neuropathic hypersensitivity in mice with conditional deletion of Npy1r from CCK-INs and NPFF-INs but not from GRP-INs. We conclude that Grp/Npy1r-INs are conserved in higher order mammalian species and represent a promising and precise pharmacotherapeutic target for the treatment of neuropathic pain.

Topics: Animals; Humans; In Situ Hybridization, Fluorescence; Interneurons; Mammals; Neuralgia; Neuropeptide Y; Peripheral Nerve Injuries

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

Neuropeptide Y (NPY) modulates nociception in the spinal cord, but little is known about its mechanisms of release. We measured NPY release in situ using the internalization of its Y1 receptor in dorsal horn neurons. Y1 receptor immunoreactivity was normally localized to the cell surface, but addition of NPY to spinal cord slices increased the number of neurons with Y1 internalization in a biphasic fashion (EC

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Arginine; Calcium Channel Blockers; Capsaicin; Cyclic AMP-Dependent Protein Kinases; Excitatory Amino Acid Antagonists; Hyperalgesia; Indoles; Interneurons; Lidocaine; Male; Microscopy, Confocal; Neuropeptide Y; Nociception; omega-Conotoxins; Peripheral Nerve Injuries; Posterior Horn Cells; Potassium Chloride; Protein Transport; Protein Tyrosine Phosphatases; Rats; Receptors, N-Methyl-D-Aspartate; Receptors, Neuropeptide Y; Sensory System Agents; Spinal Cord; Spinal Nerve Roots; TRPV Cation Channels; Voltage-Gated Sodium Channel Blockers

Heterogeneity is increasingly recognized in clinical presentation of neuropathic pain (NP), but less often recognized in animal models. Neurochemical dysregulation in rodent dorsal root ganglia (DRG) is associated with peripheral nerve trauma, but poorly studied in non-traumatic NP conditions.. This study aimed to investigate the temporal expressions of activating transcription factor-3 (ATF-3), growth-associated protein-43 (GAP-43), neuropeptide Y (NPY) and galanin in traumatic and non-traumatic rat models of neuropathies associated with NP. Expressions of these markers were examined in the DRG at different time points following tibial nerve transection (TNT) injury and antiretroviral drug stavudine (d4T) administration using immunohistochemistry. The development of sensory gain following these insults was assessed by measuring limb withdrawal to a punctate mechanical stimulus.. Both TNT-injured and d4T-treated rats developed hindpaw mechanical hypersensitivity. Robust expressions of ATF-3, GAP-43, NPY and galanin in both small- and large-sized L5 DRG neurons were observed in the DRG from TNT-injured rats. In contrast, d4T-treated rats did not exhibit any significant neurochemical changes in the DRG.. Taken together, the results suggest that ATF-3, GAP-43, NPY and galanin are likely indicators of nerve trauma-associated processes and not generic markers for NP. These experiments also demonstrate distinct expression patterns of neurochemical markers in the DRG and emphasize the mechanistic difference between nerve trauma and antiretroviral drug-associated NP.

Topics: Animals; Anti-HIV Agents; Galanin; Ganglia, Spinal; Immunohistochemistry; Male; Neurons; Neuropeptide Y; Peripheral Nerve Injuries; Rats, Wistar; Stavudine

Previous investigations of the anatomical basis of the neuropathic-like manifestations in the spinal nerve ligation animal model have shown that the central terminations of the unmyelinated primary afferents of L5 spinal nerve are not restricted to the corresponding L5 spinal segment, and rather extend to two spinal segments rostrally and one segment caudally where they intermingle with primary afferents of the adjacent L4 spinal nerve. The aim of the present study was to investigate the neurochemical changes in the dorsal horn of the spinal cord and DRGs after L5 nerve injury in rats. In the first experiment, the right L5 nerve was ligated and sectioned for 14 days, and isolectin B4 (IB4, a tracer for unmyelinated primary afferents) was injected into the left L5 nerve. The results showed that the vasoactive intestinal peptide (VIP) was up-regulated in laminae I-II of L3-L6 spinal segments on the right side in exactly the same areas where IB4 labelled terminals were revealed on the left side. In the second experiment, L5 was ligated and sectioned and the spinal cord and DRGs were stained immunocytochemically with antibodies raised against various peptides known to be involved in pain transmission and hyperalgesia. The results showed that L5 nerve lesion caused down-regulation of substance P, calcitonin-gene related peptide and IB4 binding and up-regulation of neuropeptide Y and neurokinin-1 receptor in the dorsal horn of L4 and L5 spinal segments. Similar neurochemical changes were observed only in the corresponding L5 DRG with minimal effects observed in L3, L4 and L6 DRGs. Although, L5 nerve injury caused an up-regulation in NPY, no change in SP and CGRP immunoreactivity was observed in ipsilateral garcile nucleus. These neuroplastic changes in the dorsal horn of the spinal cord, in the adjacent uninjured territories of the central terminations of the adjacent uninjured nerves, might explain the mechanism of hyperalgesia after peripheral nerve injury.

Topics: Animals; Lumbar Vertebrae; Neuralgia; Neuropeptide Y; Peripheral Nerve Injuries; Rats; Rats, Wistar; Receptors, Neurokinin-1; Spinal Cord; Spinal Nerves; Substance P; Up-Regulation

In order to investigate whether cholera toxin B subunit (CTb) is transported by unmyelinated primary afferents following nerve injury, we transected the sciatic nerves of six rats, and injected the transected nerves (and in three cases also the intact contralateral nerves) with CTb, 2 weeks later. The relationship between CTb and two neuropeptides, vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY), was then examined in neurons in the ipsilateral L4 and L5 dorsal root ganglia, using immunofluorescence staining and confocal microscopy. We also immunostained sections of spinal cord and caudal medulla for CTb, NPY and VIP. Following nerve section, VIP immunoreactivity was increased in laminae I-II of the spinal cord while NPY immunoreactivity was increased in laminae III-IV of the spinal cord and in the gracile nucleus. On the contralateral side, CTb labelling was detected in laminae I and III-V of the dorsal horn of the L4 and L5 spinal segments, as well as in the gracile nucleus. CTb labelling was seen in the same areas on the lesioned side, but with a dramatic increase in lamina II. No VIP or NPY immunoreactivity was observed in L4 and L5 dorsal root ganglia on the side of the intact nerve, but on the lesioned side VIP was detected in many small neurons and NPY in numerous large neurons. In agreement with the report by Tong et al. [J. Comp. Neurol. 404 (1999) 143], we found that while CTb labelling in the dorsal root ganglion on the side of the intact nerve was mainly in large neurons, on the lesioned side CTb was present in dorsal root ganglion neurons of all sizes. The main finding of the present study was that almost all of the VIP- (96%) and NPY- (98%) positive neurons in the dorsal root ganglia on the lesioned side were also CTb-labelled. After nerve injury VIP is upregulated in fine afferents that terminate in laminae I and II, and most of these probably have unmyelinated axons. Since the cell bodies of these neurons were labelled with CTb that had been injected into the transected sciatic nerve, this suggests that many of these fine afferents, which do not normally transport CTb, are capable of doing so after injury.

Topics: Afferent Pathways; Animals; Cholera Toxin; Fluorescent Antibody Technique; Ganglia, Spinal; Immunoenzyme Techniques; Medulla Oblongata; Microscopy, Confocal; Nerve Fibers, Unmyelinated; Neuropeptide Y; Peripheral Nerve Injuries; Peripheral Nerves; Rats; Rats, Wistar; Sciatic Nerve; Spinal Cord; Vasoactive Intestinal Peptide

Several types of changes have been reported to occur in dorsal root ganglia following peripheral nerve injury, including loss of neurons and increases and decreases in peptide expression. However, with regard to loss of neurons, results have not been consistent, presumably due to different quantitative methodologies employed and species analyzed. So far, most studies have been conducted on rats; however, with the fast development of the transgenic techniques, the mouse has become a standard model animal in primary sensory research. Therefore we used stereological methods to determine the number of neurons, as well as the expression of galanin message-associated peptide, a marker for galanin-expressing neurons, neuropeptide Y, and calcitonin gene-related peptide in lumbar 5 dorsal root ganglia of both control C57 BL/6J mice and in mice subjected to a 'mid-thigh' sciatic nerve transection (axotomy). In control animals the total number of lumbar 5 dorsal root ganglion neurons was about 12000. Seven days after axotomy, 24% of the dorsal root ganglion neurons were lost (P<0.001), and 54% were lost 28 days after axotomy (P<0.001). With regard to the percentage of peptide-expressing neurons, the results obtained showed that both galanin message-associated peptide (from <1% to about 21%) and neuropeptide Y (from <1% to about 16%) are upregulated, whereas calcitonin gene-related peptide is downregulated (from about 41% to about 14%) following axotomy. Results obtained with retrograde labeling of the axotomized dorsal root ganglion neurons indicate that the neuropeptide regulations may be even more pronounced, if the analysis is confined to the axotomized dorsal root ganglion neurons rather than including the entire neuron population. We also applied conventional profile-based counting methods to compare with the stereological data and, although the results were comparable considering the trends of changes following axotomy, the actual percentage obtained with the two methods differed markedly, both for neuropeptide Y- and, especially, for galanin message-associated peptide-positive neurons. These present results demonstrate that marked species differences exist with regard to the effect of nerve injury on dorsal root ganglion neurons. Thus, whereas no neuron loss is seen in rat up to 4 weeks after a 'mid-thigh' transection [Tandrup et al. (2000) J. Comp. Neurol. 422, 172-180], the present results indicate a dramatic loss already after 1 week in mouse. It is sugges

Topics: Animals; Axotomy; Calcitonin Gene-Related Peptide; Cell Count; Cell Death; Cell Size; Fluorescent Dyes; Galanin; Ganglia, Spinal; Immunohistochemistry; Mice; Mice, Inbred C57BL; Nerve Degeneration; Neuropeptide Y; Neuropeptides; Peripheral Nerve Injuries; Peripheral Nerves; Peripheral Nervous System Diseases; Stilbamidines

Microprobes bearing immobilized antibodies to the C-terminus of neuropeptide Y were used to measure the release of this neuropeptide in the spinal cords of rats with a unilateral peripheral neuropathy and in sham-operated animals. All neuropathic animals showed the characteristic behavioural syndrome and were studied at 14 days postsciatic nerve loose-ligation. An extensive spontaneous release of immunoreactive neuropeptide Y was detected in the spinal cords of the neuropathic rats and, compared to sham-operated rats, a new zone of release was found in the deep dorsal horn. Electrical stimulation of large diameter primary afferents proximal to the nerve ligature produced widespread release of neuropeptide Y in the dorsal horn which persisted for up to 1 h poststimulation. It is possible that ectopic impulses arising in the injured nerve were responsible for the spontaneous central release of neuropeptide Y and this neuropeptide may play a role in the central response to peripheral nerve injury.

Topics: Animals; Electric Stimulation; Immunohistochemistry; Iontophoresis; Male; Microelectrodes; Neurons, Afferent; Neuropeptide Y; Peripheral Nerve Injuries; Peripheral Nerves; Presynaptic Terminals; Rats; Rats, Wistar

Peripheral nerve transection induces significant changes in neuropeptide expression and content in injured primary sensory neurons, possibly due to loss of target derived neurotrophic support. This study shows that neurotrophin-3 (NT-3) delivery to the injured nerve influences neuropeptide Y (NPY) expression within dorsal root ganglia (DRG) neurons. NT-3 was delivered by grafting impregnated fibronectin (500 ng/ml; NT group) in the axotomised sciatic nerve. Animals grafted with plain fibronectin mats (FN) or nerve grafts (NG) were used as controls. L4 and L5 DRG from operated and contralateral sides were harvested between 5 and 240 d. Using immunohistochemistry and computerised image analysis the percentage, diameter and optical density of neurons expressing calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal peptide (VIP) and NPY were quantified. Sciatic nerve axotomy resulted in significant reduction in expression of CGRP and SP, and significant upregulation of VIP and NPY (P < 0.05 for ipsilateral vs contralateral DRG). By d 30, exogenous NT-3 and nerve graft attenuated the upregulation of NPY (P < 0.05 for NT and NG vs FN). However, NT-3 administration did not influence the expression of CGRP, SP or VIP. The mean cell diameter of NPY immunoreactive neurons was significantly smaller in the NT-3 group (P < 0.05 for NT vs FN and NG) suggesting a differential influence of NT-3 on larger neurons. The optical densities of NPY immunoreactive neurons of equal size were the same in each group at any time point, indicating that the neurons responding to NT-3 downregulate NPY expression to levels not detectable by immunohistochemistry. These results demonstrate that targeted administration of NT-3 regulates the phenotype of a NPY-immunoreactive neuronal subpopulation in the dorsal root ganglia, a further evidence of the trophic role of neurotrophins on primary sensory neurons.

Topics: Analysis of Variance; Animals; Calcitonin Gene-Related Peptide; Fibronectins; Ganglia, Spinal; Image Processing, Computer-Assisted; Immunohistochemistry; Male; Nerve Growth Factors; Nerve Regeneration; Neurons, Afferent; Neuropeptide Y; Neurotrophin 3; Peripheral Nerve Injuries; Peripheral Nerves; Rats; Rats, Inbred Lew; Substance P; Time Factors; Vasoactive Intestinal Peptide

Combined retrograde neuronal tracing with FluoroGold (FG) and a double immunofluorescence method was performed to examine the effects of peripheral nerve injury of the masseteric nerve (MassN) on the levels of two calcium binding proteins (CaBPs), parvalbumin (PV) and calbindin D28k (CB), and neuropeptide Y (NPY) in the mesencephalic trigeminal nucleus (MesV) in the rat. In the normal MesV, many medium- to large-sized unipolar PV-like immunoreactive (-IR) cells were detected through the entire rostrocaudal extent, but CB-IR cells were rarely observed. No NPY-IR cells were observed in the normal MesV. The distributions of these three neurochemical markers in the MesV contralateral to the transection of Mass were almost identical to those observed in the normal MesV. Four days following transection and application of FG to the MassN, approximately 52% (572/1104) and 38% (414/1104) of FG-labeled cells (FG cells) in the MesV displayed PV-like immunoreactivity (-LI) and NPY-LI, respectively; Approximately 24% (265/1104) of FG cells showed both PV-LI and NPY-LI. Approximately 47% (265/572) of FG cells with PV-LI showed NPY-LI or 64% (265/414) of FG cells with NPY-LI displayed PV-LI. Fourteen days following transection and application of FG, the percentage of FG cells with PV-LI significantly decreased to 36% (365/1024) compared to that observed 4 days post-injury; approximately 44% (448/1024) of FG cells displayed NPY-LI; approximately 38% (141/365) of FG cells with PV-LI showed NPY-LI and approximately 31% (141/448) of FG cells with NPY-LI displayed PV-LI. In contrast, FG cells showing CB-LI were very rare on 4 days (1%; 15/1182) or 14 days (1%; 16/1085) following MassN transection. The present results indicate that the levels of PV in the MesV decreased 14 days following the MassN injury compared to those observed 4 days post-injury and rapid induction of NPY in the injured MesV neurons, and that the correlation between CaBP and NPY in the MesV following the MassN transection is different from that observed in the trigeminal ganglion, which is equivalent to the MesV, following peripheral nerve injury of the inferior alveolar nerve.

Topics: Animals; Calbindin 1; Calbindins; Calcium-Binding Proteins; Denervation; Male; Masseter Muscle; Mesencephalon; Neuropeptide Y; Parvalbumins; Peripheral Nerve Injuries; Rats; Rats, Sprague-Dawley; Reference Values; S100 Calcium Binding Protein G; Trigeminal Nuclei

By in situ hybridization and immunohistochemistry, we examined the expression of neuropeptides such as neuropeptide Y (NPY), galanin (Gal), substance P (SP), vasoactive intestinal polypeptide (VIP) and their mRNAs in the rat mesencephalic trigeminal nucleus (Mes5) following masseteric nerve transection. On the side contralateral to the nerve transection, none of the peptides examined were labeled in Mes5 cell bodies. However, on the side ipsilateral to the lesion, NPY, Gal and preprotachykinin (PPT) mRNAs appeared in Mes5 cell bodies. Double labeling for mRNAs by in situ hybridization and retrograde tracer fluoro-gold (FG) revealed that almost all (96-97%) the FG-labeled neurons which were cut expressed NPY and Gal mRNAs, whereas less neurons (87%) expressed PPT mRNA. NPY and Gal-like immunoreactivities were detected in Mes5 cell bodies ipsilateral to the axotomy. The results suggested that these neuropeptides play roles in adaptive processes after peripheral nerve injury in Mes5 neurons as they are thought to do so in dorsal root ganglion neurons.

Topics: Animals; Galanin; In Situ Hybridization; Male; Masseter Muscle; Mesencephalon; Neuropeptide Y; Neuropeptides; Peptide Biosynthesis; Peptides; Peripheral Nerve Injuries; Rats; Rats, Sprague-Dawley; RNA, Messenger; Substance P; Trigeminal Nerve; Vasoactive Intestinal Peptide