scholarly journals CGRP measurements in human plasma – a methodological study

Cephalalgia ◽  
2021 ◽  
pp. 033310242110241
Author(s):  
Karl Messlinger ◽  
Birgit Vogler ◽  
Annette Kuhn ◽  
Julika Sertel-Nakajima ◽  
Florian Frank ◽  
...  
Keyword(s):  
Plasma Levels ◽  
Calcitonin Gene Related Peptide ◽  
Enzyme Linked Immunosorbent Assay ◽  
Published Data ◽  
Human Blood Plasma ◽  
Primary Headaches ◽  
Peptidase Inhibitors ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Standard Solutions

Background Calcitonin gene-related peptide plasma levels have frequently been determined as a biomarker for primary headaches. However, published data is often inconsistent resulting from different methods that are not precisely described in most studies. Methods We applied a well-proven enzyme-linked immunosorbent assay to measure calcitonin gene-related peptide concentrations in human blood plasma, we modified parameters of plasma preparation and protein purification and used calcitonin gene-related peptide-free plasma for standard solutions, which are described in detail. Results Calcitonin gene-related peptide levels are stable in plasma with peptidase inhibitors and after deep-freezing. Calcitonin gene-related peptide standard solutions based on synthetic intercellular fluid or pooled plasma with pre-absorbed calcitonin gene-related peptide influenced the measurements but yielded both comprehensible results. In a sample of 56 healthy subjects the calcitonin gene-related peptide plasma levels varied considerably from low (<50 pg/mL) to very high (>500 pg/mL) values. After a 12-hour exposure of these subjects to normobaric hypoxia the individual calcitonin gene-related peptide levels remained stable. Conclusion Buffering with peptidase inhibitors and immediate freezing or processing of plasma samples is essential to achieve reliable measurements. Individuals show considerable differences and partly high calcitonin gene-related peptide plasma levels without detectable pathological reason. Thus plasma measurements are suited particularly to follow calcitonin gene-related peptide levels in longitudinal studies. The use of data for this study was approved by the Ethics Committee of the Medical University of Innsbruck ( https://www.i-med.ac.at/ethikkommission/ ; EK Nr: 1242/2017).

scholarly journals Calcitonin Gene-Related Peptide in Tension-Type Headache

2002 ◽  
Vol 2 ◽  
pp. 1527-1531 ◽  
Author(s):  
M. Ashina
Keyword(s):  
Plasma Levels ◽  
Calcitonin Gene Related Peptide ◽  
Tension Type Headache ◽  
Nerve Fibers ◽  
Primary Headaches ◽  
Chronic Tension Type Headache ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Type Headache

In the last 10 years there has been increasing interest in the role of calcitonin gene-related peptide (CGRP) in primary headaches. Tension-type headache is one of the most common and important types of primary headaches, and ongoing nociception from myofascial tissues may play an important role in the pathophysiology of this disorder. CGRP sensory fibers are preferentially located in the walls of arteries, and nerve fibers containing CGRP accompany small blood vessels in human cranial muscles. It is well established that nociception may lead to release of CGRP from sensory nerve endings and from central terminals of sensory afferents into the spinal cord. It has also been shown that density of CGRP fibers around arteries is increased in persistently inflamed muscle. These findings indicate that ongoing activity in sensory neurons in the cranial muscles may be reflected in changes of plasma levels of neuropeptides in patients with chronic tension-type headache. To explore the possible role of CGRP in tension-type headache, plasma levels of CGRP were measured in patients with chronic tension-type headache. This study showed that plasma levels of CGRP are normal in patients and unrelated to headache state. However, the findings of normal plasma levels of CGRP do not exclude the possibility that abnormalities of this neuropeptide at the neuronal or peripheral (pericranial muscles) levels play a role in the pathophysiology of tension-type headache. Investigation of CGRP in other compartments with new sensitive methods of analysis is necessary to clarify its role in tension-type headache.

Plasma levels of calcitonin gene-related peptide in chronic tension-type headache

Neurology ◽  
2000 ◽  
Vol 55 (9) ◽  
pp. 1335-1340 ◽  
Author(s):  
M. Ashina ◽  
L. Bendtsen ◽  
R. Jensen ◽  
S. Schifter ◽  
I. Jansen-Olesen ◽  
...  
Keyword(s):  
Plasma Levels ◽  
Calcitonin Gene Related Peptide ◽  
Tension Type Headache ◽  
Chronic Tension Type Headache ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Type Headache

Calcitonin gene-related peptide elevates calcium and polarizes membrane potential in MG-63 cells by both cAMP-independent and -dependent mechanisms

2004 ◽  
Vol 287 (2) ◽  
pp. C457-C467 ◽  
Author(s):  
Douglas M. Burns ◽  
Lisa Stehno-Bittel ◽  
Tomoyuki Kawase
Keyword(s):  
Membrane Potential ◽  
Signaling Pathways ◽  
Secondary Phase ◽  
Calcitonin Gene Related Peptide ◽  
Published Data ◽  
Maximal Effect ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Intracellular Ca ◽  
20 Nm

Published data suggest that the neuropeptide calcitonin gene-related peptide (CGRP) can stimulate osteoblastic bone formation; however, interest has focused on activation of cAMP-dependent signaling pathways in osteogenic cells without full consideration of the importance of cAMP-independent signaling. We have now examined the effects of CGRP on intracellular Ca2+ concentration ([Ca2+]int) and membrane potential ( Em) in preosteoblastic human MG-63 cells by single-cell fluorescent confocal analysis using fluo 4-AM-fura red-AM and bis(1,3-dibarbituric acid)-trimethine oxanol [DiBAC4( 3 )] bis-oxonol assays. CGRP produced a two-stage change in [Ca2+]int: a rapid transient peak and a secondary sustained increase. Both responses were dose dependent with an EC50 of ∼0.30 nM, and the maximal effect (initially ∼3-fold over basal levels) was observed at 20 nM. The initial phase was sensitive to inhibition of Ca2+ mobilization with thapsigargin, whereas the secondary phase was eliminated only by blocking transmembrane Ca2+ influx with verapamil or inhibiting cAMP-dependent signaling with the Rp isomer of adenosine 3′,5′-cyclic monophosphorothioate (Rp-cAMPS). These data suggest that CGRP initially stimulates Ca2+ discharge from intracellular stores by a cAMP-independent mechanism and subsequently stimulates Ca2+ influx through L-type voltage-dependent Ca2+ channels by a cAMP-dependent mechanism. In addition, CGRP dose-dependently polarized cellular Em, with maximal effect at 20 nM and an EC50 of 0.30 nM. This effect was attenuated with charybdotoxin (−20%) or glyburide (glibenclamide; −80%), suggesting that Em hyperpolarization is induced by both Ca2+-activated and ATP-sensitive K+ channels. Thus CGRP signals strongly by both cAMP-dependent and cAMP-independent signaling pathways in preosteoblastic human MG-63 cells.

Differences in pituitary adenylate cyclase-activating peptide and calcitonin gene-related peptide release in the trigeminovascular system

Cephalalgia ◽  
2020 ◽  
Vol 40 (12) ◽  
pp. 1296-1309 ◽  
Author(s):  
Jacob Carl Alexander Edvinsson ◽  
Anne-Sofie Grell ◽  
Karin Warfvinge ◽  
Majid Sheykhzade ◽  
Lars Edvinsson ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Trigeminal Ganglion ◽  
Calcitonin Gene Related Peptide ◽  
Receptor Subtype ◽  
Primary Headaches ◽  
Satellite Glial Cells ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Pituitary Adenylate Cyclase ◽  
Peptide Release

Background Several neurotransmitters are expressed in the neurons of the trigeminal ganglion. One such signalling molecule is the pituitary adenylate cyclase-activating peptide (PACAP). PACAP signalling has been suggested to have a possible role in the pathophysiology of primary headaches. Objective The present study was designed to investigate the relationship between PACAP and calcitonin gene-related peptide, currently the two most relevant migraine peptides. Methods In the current study, we used ELISA to investigate PACAP and calcitonin gene-related peptide release in response to 60 mM K+ or capsaicin using a rat hemi-skull model. We combined this analysis with qPCR and immunohistochemistry to study the expression of PACAP and calcitonin gene-related peptide receptors and ligands. Results Calcitonin gene-related peptide (CGRP) is released from the trigeminal ganglion and dura mater. In contrast, PACAP is only released from the trigeminal ganglion. We observed a weak correlation between the stimulated release of the two neuropeptides. PACAP-38 immunoreactivity was expressed alone and in a subpopulation of neurons in the trigeminal ganglion that also store calcitonin gene-related peptide. The receptor subtype PAC1 was mainly expressed in the satellite glial cells (SGCs), which envelop the neurons in the trigeminal ganglion, in some neuronal processes, inside the Aδ-fibres and in the outermost layer of the myelin sheath that envelopes the Aδ-fibres. Conclusion Unlike CGRP, PACAP is only released within the trigeminal ganglion. This raises the question of whether a migraine therapy aimed at preventing peripheral PACAP signalling would be as successful as the CGRP signalling targeted treatments.

Cellular distribution of PACAP-38 and PACAP receptors in the rat brain: Relation to migraine activated regions

Cephalalgia ◽  
2019 ◽  
Vol 40 (6) ◽  
pp. 527-542 ◽  
Author(s):  
Karin Warfvinge ◽  
Lars Edvinsson
Keyword(s):  
Amino Acid ◽  
Rat Brain ◽  
Vasoactive Intestinal Polypeptide ◽  
Calcitonin Gene Related Peptide ◽  
Amino Acid Sequences ◽  
Primary Headaches ◽  
Functional Responses ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Intestinal Polypeptide

Background Pituitary adenylate cyclase-activating polypeptide (PACAP) occurs as either a 27- or 38-amino acid neuropeptide and belongs to the vasoactive intestinal polypeptide/glucagon/secretin family of peptides. PACAP and vasoactive intestinal polypeptide have a 68% homology of their amino acid sequences and share three B-type G-protein coupled receptors: VPAC1, VPAC2 and PAC1 receptors. Methods/results The distribution of PACAP-38 and its receptors in the brain is only partly described in the literature. Here, we have performed a study to provide the more general picture of this system in rat brain in order to understand a putative role in primary headaches and partly in relation to the calcitonin gene-related peptide system. We observed a rich expression of PACAP-38 and PAC1 receptor immunoreactivity in many regions throughout the cerebrum, cerebellum and brainstem. The expression pattern points to multiple functions, not least associated with pain and reactions to pain. The expression of VPAC1 and VPAC2 receptor immunoreactivity was very sparse. In several regions such as the cerebral cortex, trigeminal nucleus caudalis, hypothalamus and pons there was a close relation to calcitonin gene-related peptide expression. Conclusion The findings suggest that the rich supply of PACAP-38 and PAC1 receptors is associated with basic functional responses in the central nervous system (CNS), and there are important close anatomical relations with calcitonin gene-related peptide in CNS regions associated with migraine pathophysiology.

Ecabet sodium raises plasma levels of calcitonin gene-related peptide and substance P in healthy humans

2005 ◽  
Vol 57 (6) ◽  
pp. 799-805 ◽  
Author(s):  
Fumihiko Katagiri ◽  
Shin Inoue ◽  
Yuhki Sato ◽  
Hiroki Itoh ◽  
Masaharu Takeyama
Keyword(s):  
Substance P ◽  
Plasma Levels ◽  
Calcitonin Gene Related Peptide ◽  
Ecabet Sodium ◽  
Related Peptide ◽  
Healthy Humans ◽  
Calcitonin Gene

scholarly journals Calcitonin gene–related peptide inhibits Langerhans cell–mediated HIV-1 transmission

2013 ◽  
Vol 210 (11) ◽  
pp. 2161-2170 ◽  
Author(s):  
Yonatan Ganor ◽  
Anne-Sophie Drillet-Dangeard ◽  
Lucia Lopalco ◽  
Daniela Tudor ◽  
Giuseppe Tambussi ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Highly Active Antiretroviral Therapy ◽  
Plasma Levels ◽  
Calcitonin Gene Related Peptide ◽  
Cell Infection ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Hiv 1

Upon its mucosal entry, human immunodeficiency virus type 1 (HIV-1) is internalized by Langerhans cells (LCs) in stratified epithelia and transferred locally to T cells. In such epithelia, LCs are in direct contact with peripheral neurons secreting calcitonin gene–related peptide (CGRP). Although CGRP has immunomodulatory effects on LC functions, its potential influence on the interactions between LCs and HIV-1 is unknown. We show that CGRP acts via its receptor expressed by LCs and interferes with multiple steps of LC-mediated HIV-1 transmission. CGRP increases langerin expression, decreases selected integrins, and activates NF-κB, resulting in decreased HIV-1 intracellular content, limited formation of LC–T cell conjugates, and elevated secretion of the CCR5-binding chemokine CCL3/MIP-1α. These mechanisms cooperate to efficiently inhibit HIV-1 transfer from LCs to T cells and T cell infection. In vivo, HIV-1 infection decreases CGRP plasma levels in both vaginally SHIV-challenged macaques and HIV-1–infected individuals. CGRP plasma levels return to baseline after highly active antiretroviral therapy. Our results reveal a novel path by which a peripheral neuropeptide acts at the molecular and cellular levels to limit mucosal HIV-1 transmission and suggest that CGRP receptor agonists might be used therapeutically against HIV-1.

scholarly journals Comparative evaluation of serum calcitonin gene related peptide level in patients with migraine headache with and without aura

2021 ◽  
Author(s):  
Mostafa Rezaee ◽  
Nahid Ashja zadeh ◽  
Sadegh Izedi ◽  
Farinaz Fakhri
Keyword(s):  
Correlation Coefficient ◽  
Rank Correlation ◽  
Calcitonin Gene Related Peptide ◽  
Clinical Findings ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Serum Calcitonin ◽  
Related Peptide ◽  
Spearman’S Rank Correlation ◽  
Calcitonin Gene

Abstract Background During a migraine attack, trigeminal activation results in the release of calcitonin gene-related peptide (CGRP), which stimulates the release of inflammatory cytokines playing an important role in migraine. We analyze the serum level of CGRP between two groups of migrainous patients (with aura and without aura) Materials and Methods Thirty six migraine patients (included 18 patients with aura and 18 without aura) additionally 18 healthy volunteers consisted control group were selected from the clinic of Neurology, Shiraz University of Medical Sciences, Shiraz, Iran, between March 2020 and November 2020. The CGRP level were determined from the sera of patients with migraine and control subjects by enzyme-linked immunosorbent assay kits. Spearman's rank correlation coefficient was also determined to calculate the correlation between CGRP and clinical findings. Results The level of CGRP in groups were significantly different between groups (P = 0.00). Also, the level of CGRP in aura group were significantly higher than non-aura group (P = 0.045). The Spearman’s correlation coefficient revealed a positive and significant correlation between the CGRP concentration and age (p = 0.042, r = 0.172), BMI (p = 0.013, r = 0.08), VAS (P = 0.006 ,r = 0.09), frequency of attacks (p = 0.005, r = 0.9), duration of each attack (p = 0.016, r = 0.23), Migraine Disability Assessment Scale.(p = 0.00, r = 0.785), average of number of Medication (p = 0.00, r = 0.694). However, no significant correlation was observed with gender. (P > 0.05 ) Conclusions In our study, we found migraine patients had a higher CGRP level than healthy controls and the level of CGRP was related significantly with the duration, BMI, frequency of headache, age, number of headaches per day. In conclusion, our results confirmed that CGRP may be involved in the pathogenesis of migraine attacks and related with the multiple clinical characteristics.

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