scholarly journals Carbocyanine Postmortem Neuronal Tracing: Influence of Different Parameters on Tracing Distance and Combination with Immunocytochemistry

1998 ◽  
Vol 46 (8) ◽  
pp. 901-910 ◽  
Author(s):  
Julius-Robert Lukas ◽  
Martin Aigner ◽  
Michaela Denk ◽  
Harald Heinzl ◽  
Martin Burian ◽  
...  
Keyword(s):  
Cervical Spinal Cord ◽  
Trigeminal Ganglia ◽  
Detailed Data ◽  
Animal Tissues ◽  
Neuronal Tracing ◽  
Calcitonin Gene ◽  
Sciatic Nerves ◽  
Neuronal Tracers ◽  
Incubation Temperatures ◽  
Newborn Animals

SUMMARY Carbocyanines (DiI, DiA, DiO) are able to travel along membranes by diffusion and therefore have been used as postmortem neuronal tracers in aldehyde-fixed tissues. Surprisingly, detailed data on the influence of different parameters on tracing distances are still missing. This study was carried out to optimize tracing procedures and to reveal the validity of the combination of postmortem tracing with immunocytochemistry. Carbocyanine crystals were applied to the cervical spinal cord, sciatic nerves, and brachial plexuses of humans and guinea pigs. Incubation in the dark at 37C for 12-15 weeks proved optimal to achieve longest tracing distances (28.9 ± 2.2 mm) in human and animal tissues. Longer incubation times and incubation temperatures higher than 37C did not result in longer tracing distances. No differences were evident between adult and newborn animals and between central and peripheral nervous system. The diffusion coefficient for DiI was calculated to be 2.5 × 10-7 cm2sec-1. After application of DiI to nerves of guinea pig extraocular muscles, DiI-positive afferent perikarya were observed in the anteromedial part of the trigeminal ganglion. These perikarya were identified by calcitonin gene-related peptide immunoreactivity (CGRP-IR). The percentage of CGRP-IR neurons after tracing was concordant with the percentage of CGRP-IR in trigeminal ganglia exclusively processed for CGRP-IR without previous postmortem tracing. These results demonstrate carbocyanines to be specific tracers for exact neuronal mapping studies.

scholarly journals Targeted Regulation Of Cgrp Gene Expression

2001 ◽  
Vol 1 ◽  
pp. 5-5 ◽  
Author(s):  
A. F. Russo ◽  
P. L. Durham
Keyword(s):  
Map Kinase ◽  
Therapeutic Strategy ◽  
Adenoviral Vectors ◽  
Calcium Signal ◽  
Trigeminal Ganglia ◽  
Cerebral Vasculature ◽  
Trigeminal Neuron ◽  
Calcitonin Gene ◽  
Gene Enhancer ◽  
A Cell

The neuropeptide calcitonin gene-related peptide (CGRP) is a potent regulator of cerebral vascular tone and contributes to neurogenic inflammation. Clinical studies have shown that CGRP levels are elevated during the painful phase of migraine headache, then restored to baseline by antimigraine 5-HT1 drugs. Conversely, CGRP is depleted in perivascular nerve terminals from patients who have suffered vasospasm following subarachnoid hemorrhage. We have investigated the mechanisms controlling CGRP expression in the trigeminal ganglia neurons, which provide virtually all of the CGRP innervation to the cerebral vasculature. We found that nerve depolarization, inflammatory compounds, and nitric oxide can increase CGRP synthesis and secretion. Using both adenoviral vectors and transfection approaches, we have shown that the increased synthesis is due to activation of a cell-specific MAP kinase-responsive enhancer upstream of the CGRP gene. Interestingly, the 5-HT1 migraine drugs are able to block this up-regulation by a mechanism that involves a very prolonged elevation of calcium. We have shown that the duration of the calcium signal is a key determinant for whether a MAP kinase responsive gene will be stimulated or repressed by calcium-activated pathways. This observation supports the importance of a finely tuned balance of calcium in the trigeminal neuron, which is intriguing in light of genetic evidence for calcium channel mutations in a rare form of inherited migraine. These studies suggest that modulation of MAP kinase control of the cell-specific CGRP gene enhancer may be a useful therapeutic strategy for neurovascular disorders.

Oxytocin as a regulatory neuropeptide in the trigeminovascular system: Localization, expression and function of oxytocin and oxytocin receptors

Cephalalgia ◽  
2020 ◽  
Vol 40 (12) ◽  
pp. 1283-1295
Author(s):  
Karin Warfvinge ◽  
Diana N Krause ◽  
Aida Maddahi ◽  
Anne-Sofie Grell ◽  
Jacob CA Edvinsson ◽  
...  
Keyword(s):  
Dura Mater ◽  
Calcitonin Gene Related Peptide ◽  
Trigeminal Ganglia ◽  
Trigeminal Nucleus ◽  
Trigeminovascular System ◽  
Related Peptide ◽  
Oxytocin Receptors ◽  
Calcitonin Gene ◽  
And Function ◽  
Cranial Arteries

Background Recent clinical findings suggest that oxytocin could be a novel treatment for migraine. However, little is known about the role of this neuropeptide/hormone and its receptor in the trigeminovascular pathway. Here we determine expression, localization, and function of oxytocin and oxytocin receptors in rat trigeminal ganglia and targets of peripheral (dura mater and cranial arteries) and central (trigeminal nucleus caudalis) afferents. Methods The methods include immunohistochemistry, messenger RNA measurements, quantitative PCR, release of calcitonin gene-related peptide and myography of arterial segments. Results Oxytocin receptor mRNA was expressed in rat trigeminal ganglia and the receptor protein was localized in numerous small to medium-sized neurons and thick axons characteristic of A∂ sensory fibers. Double immunohistochemistry revealed only a small number of neurons expressing both oxytocin receptors and calcitonin gene-related peptide. In contrast, double immunostaining showed expression of the calcitonin gene-related peptide receptor component receptor activity-modifying protein 1 and oxytocin receptors in 23% of the small cells and in 47% of the medium-sized cells. Oxytocin immunofluorescence was observed only in trigeminal ganglia satellite glial cells. Oxytocin mRNA was below detection limit in the trigeminal ganglia. The trigeminal nucleus caudalis expressed mRNA for both oxytocin and its receptor. K+-evoked calcitonin gene-related peptide release from either isolated trigeminal ganglia or dura mater and it was not significantly affected by oxytocin (10 µM). Oxytocin directly constricted cranial arteries ex vivo (pEC50 ∼ 7); however, these effects were inhibited by the vasopressin V1A antagonist SR49059. Conclusion Oxytocin receptors are extensively expressed throughout the rat trigeminovascular system and in particular in trigeminal ganglia A∂ neurons and fibers, but no functional oxytocin receptors were demonstrated in the dura and cranial arteries. Thus, circulating oxytocin may act on oxytocin receptors in the trigeminal ganglia to affect nociception transmission. These effects may help explain hormonal influences in migraine and offer a novel way for treatment.

Expression of calcitonin gene-related peptide1 receptor mRNA in human trigeminal ganglia and cerebral arteries

1997 ◽  
Vol 229 (3) ◽  
pp. 209-211 ◽  
Author(s):  
Lars Edvinsson ◽  
Leonor Cantera ◽  
Inger Jansen-Olesen ◽  
Rolf Uddman
Keyword(s):  
Cerebral Arteries ◽  
Trigeminal Ganglia ◽  
Receptor Mrna ◽  
Calcitonin Gene

scholarly journals Effects of NSAIDs on the Release of Calcitonin Gene-Related Peptide and Prostaglandin E2from Rat Trigeminal Ganglia

2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Vittorio Vellani ◽  
Giorgia Moschetti ◽  
Silvia Franchi ◽  
Chiara Giacomoni ◽  
Paola Sacerdote ◽  
...  
Keyword(s):  
Oral Administration ◽  
Calcitonin Gene Related Peptide ◽  
Prostaglandin E ◽  
Trigeminal Ganglia ◽  
Related Peptide ◽  
Cgrp Release ◽  
Inflammatory Soup ◽  
Calcitonin Gene ◽  
Rapid Effect ◽  
Inflammatory Drugs

Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used to treat migraine, but the mechanisms of their effects in this pathology are not fully elucidated. The trigeminal ganglia and calcitonin gene-related peptide (CGRP) have been implicated in the pathophysiology of migraine. The release of CGRP and prostaglandin E2(PGE2) from freshly isolated rat trigeminal ganglia was evaluated after oral administration of nimesulide, etoricoxib, and ketoprofen, NSAIDs with different pharmacological features. Thirty minutes after oral administration, nimesulide, 10 mg/Kg, decreased the GCRP release induced by an inflammatory soup, while the other NSAIDs were ineffective at this point in time. Two hours after oral nimesulide (5 and 10 mg/Kg) and ketoprofen (10 mg/Kg), but not of etoricoxib, a significant decrease in the CGRP release was observed. All drugs reduced PGE2, although with some differences in timing and doses, and the action on CGRP does not seem to be related to PGE2inhibition. The reduction of CGRP release from rat trigeminal ganglia after nimesulide and ketoprofen may help to explain the mechanism of action of NSAIDs in migraine. Since at 30 minutes only nimesulide was effective in reducing CGRP release, these results suggest that this NSAID may exert a particularly rapid effect in patients with migraine.

scholarly journals N/OFQ modulates orofacial pain induced by tooth movement through CGRP-dependent pathways

2020 ◽  
Author(s):  
Xinyu Yan ◽  
Han Han ◽  
Shizhen Zhang ◽  
Yanzhu Lu ◽  
Linghuan Ren ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Tooth Movement ◽  
Orofacial Pain ◽  
Pain Modulation ◽  
Trigeminal Ganglia ◽  
Orphanin Fq ◽  
Pain Model ◽  
Calcitonin Gene ◽  
Agonist And Antagonist

Abstract Background: Nociceptin/orphanin FQ (N/OFQ) has been revealed to play bidirectional roles in orofacial pain modulation. Calcitonin gene-related peptide (CGRP) is a well-known pro-nociceptive molecule that participates in the modulation of orofacial pain. We aimed to determine the effects of N/OFQ on the modulation of orofacial pain and on the release of CGRP.Methods: Orofacial pain model was established by ligating springs between incisors and molars in rats for the simulation of tooth movement. The expression level of N/OFQ was determined and pain level scored in response to orofacial pain. Both agonist and antagonist of N/OFQ receptor were administered to examine their effects on pain and the expression of CGRP in trigeminal ganglia (TG). Moreover, gene therapy based on the overexpression of N/OFQ was delivered to validate the modulatory role of N/OFQ on pain and CGRP expression.Results: Tooth movement elicited orofacial pain and an elevation in N/OFQ expression. N/OFQ exacerbated orofacial pain and upregulated CGRP expression in TG, while UFP-101 alleviated pain and downregulated CGRP expression. N/OFQ-based gene therapy was successful in overexpressing N/OFQ in TG, which resulted in pain exacerbation and elevation of CGRP expression in TG.Conclusions: N/OFQ exacerbated orofacial pain possibly through upregulating CGRP.

scholarly journals N/OFQ modulates orofacial pain induced by tooth movement through CGRP-dependent pathways

2020 ◽  
Author(s):  
Xinyu Yan ◽  
Han Han ◽  
Shizhen Zhang ◽  
Yanzhu Lu ◽  
Linghuan Ren ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Tooth Movement ◽  
Orofacial Pain ◽  
Pain Modulation ◽  
Trigeminal Ganglia ◽  
Orphanin Fq ◽  
Pain Model ◽  
Calcitonin Gene ◽  
Agonist And Antagonist

Abstract Background: Nociceptin/orphanin FQ (N/OFQ) has been revealed to play bidirectional roles in orofacial pain modulation. Calcitonin gene-related peptide (CGRP) is a well-known pro-nociceptive molecule that participates in the modulation of orofacial pain. We aimed to determine the effects of N/OFQ on the modulation of orofacial pain and on the release of CGRP.Methods: Orofacial pain model was established by ligating springs between incisors and molars in rats for the simulation of tooth movement. The expression level of N/OFQ was determined and pain level scored in response to orofacial pain. Both agonist and antagonist of N/OFQ receptor were administered to examine their effects on pain and the expression of CGRP in trigeminal ganglia (TG). Moreover, gene therapy based on the overexpression of N/OFQ was delivered to validate the modulatory role of N/OFQ on pain and CGRP expression.Results: Tooth movement elicited orofacial pain and an elevation in N/OFQ expression. N/OFQ exacerbated orofacial pain and upregulated CGRP expression in TG, while UFP-101 alleviated pain and downregulated CGRP expression. N/OFQ-based gene therapy was successful in overexpressing N/OFQ in TG, which resulted in pain exacerbation and elevation of CGRP expression in TG.Conclusions: N/OFQ exacerbated orofacial pain possibly through upregulating CGRP.

scholarly journals Neuropeptide Y Y1 Receptor Effects on Pulpal Nociceptors

2008 ◽  
Vol 87 (10) ◽  
pp. 948-952 ◽  
Author(s):  
J.L. Gibbs ◽  
K.M. Hargreaves
Keyword(s):  
Neuropeptide Y ◽  
Sensory Neurons ◽  
Dental Pulp ◽  
Dental Pain ◽  
Trigeminal Ganglia ◽  
Y1 Receptor ◽  
Cgrp Release ◽  
Calcitonin Gene ◽  
Peptide Release ◽  
Physiological Systems

Neuropeptide Y (NPY) is an important modulatory neuropeptide that regulates several physiological systems, including the activity of sensory neurons. We evaluated whether activation of the NPY Y1 receptor could modulate the activity of capsaicin-sensitive nociceptors in trigeminal ganglia and dental pulp. We tested this hypothesis by measuring capsaicin-stimulated calcitonin gene-related peptide release (CGRP) as a measure of nociceptor activity. Capsaicin-evoked CGRP release was inhibited by 50% (p < 0.05) in trigeminal ganglia and by 26% (p < 0.05) in dental pulp when tissues were pre-treated with [Leu31,Pro34]NPY. The Y1 receptor was found to co-localize with the capsaicin receptor TRPV1 in trigeminal ganglia. These results demonstrate that activation of the Y1 receptor results in the inhibition of the activity of capsaicin-sensitive nociceptors in the trigeminal ganglia and dental pulp. These findings are relevant to the physiological modulation of dental nociceptors by endogenous NPY and demonstrate an important novel analgesic target for the treatment of dental pain.

Functional Calcitonin Gene-Related Peptide Type 1 and Adrenomedullin Receptors in Human Trigeminal Ganglia, Brain Vessels, and Cerebromicrovascular or Astroglial Cells in Culture

1999 ◽  
Vol 19 (11) ◽  
pp. 1270-1278 ◽  
Author(s):  
Maria Jesús Moreno ◽  
Zvi Cohen ◽  
Danica B. Stanimirovic ◽  
Edith Hamel
Keyword(s):  
Human Brain ◽  
Receptor Antagonist ◽  
Cell Cultures ◽  
Calcitonin Gene Related Peptide ◽  
Trigeminal Ganglia ◽  
Astroglial Cells ◽  
Brain Vessels ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Camp Formation

Calcitonin gene-related peptide (CGRP) and adrenomedullin (ADM) are potent dilators of human brain arteries, and they have been implicated in the neurogenic inflammation underlying migraine headache and in the evolution of stroke, respectively. However, little is known about the presynaptic and postsynaptic distribution of their respective receptors in the human cerebrovascular bed and trigeminovascular system. In the current study, the expression of mRNA for ADM and the two cloned human CGRP1 receptors (identified here as A-CGRP1 receptors [ Aiyar et al., 1996 ] and K-CGRP1 receptors) [ Kapas and Clark, 1995 ] were evaluated in human brain vessels and trigeminal ganglia. Further, the ability of CGRP and ADM to activate adenylate cyclase in cerebromicrovascular and astroglial cell cultures was determined, and the receptors involved were characterized pharmacologically. Isolated human pial vessels, intracortical microvessels, and capillaries, as well as cultures of brain endothelial (EC), smooth muscle (SMC), and astroglial (AST) cells, all expressed mRNA for the two cloned CGRP1 receptors; however, message for the K-CGRP1 receptor was barely detectable in microvascular tissues and cells. In contrast, only isolated capillaries and cultured AST exhibited message for the ADM receptor. In human trigeminal ganglia, mRNA for ADM and the two CGRP1 receptors was systematically present The CGRP dose-dependently increased (up to 50-fold) cAMP formation in cell cultures, an effect significantly blocked by 0.1 to 10 μmol/L of the CGRP1 receptor antagonist CGRP8–37. The ADM receptor agonist, ADM13–52 (1 μmol/L), similarly increased cAMP production in all cell types, and this response was virtually abolished by 1 μmol/L CGRP8–37. Low concentrations (1 to 10 μmol/L) of the ADM receptor antagonist ADM22–52 blocked the ADM13–52-induced cAMP formation in AST (26% at 10 μmol/L, P < 0.05), whereas they potentiated this response in brain EC and SMC (40% and 100%, P < 0.001, respectively). Even at a higher dose (50 μmol/L), ADM22–52 inhibited the ADM13–52 effect in vascular cells (45%) much less effectively than in AST (95%). These results indicate that both CGRP and ADM can affect human brain vessels through a CGRP1 receptor, and they further suggest the presence of functional ADM receptors in human astroglial cells.

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