Visualizing the Calcitonin Gene-Related Peptide Immunoreactive Innervation of the Rat Cranial Dura Mater with Immunofluorescence and Neural Tracing

Calcitonin gene-related peptide (CGRP) is regarded as a key mediator in the generation of primary headaches. CGRP receptor antagonists reduce migraine pain in clinical trials and spinal trigeminal activity in animal experiments. The site of CGRP receptor inhibition causing these effects is debated. Activation and inhibition of CGRP receptors in the trigeminal ganglion may influence the activity of trigeminal afferents and hence of spinal trigeminal neurons. In anesthetized rats extracellular activity was recorded from neurons with meningeal afferent input in the spinal trigeminal nucleus caudalis. Mechanical stimuli were applied at regular intervals to receptive fields located in the exposed cranial dura mater. α-CGRP (10−5 M), the CGRP receptor antagonist olcegepant (10−3 M), or vehicle was injected through the infraorbital canal into the trigeminal ganglion. The injection of volumes caused transient discharges, but vehicle, CGRP, or olcegepant injection was not followed by significant changes in ongoing or mechanically evoked activity. In animals pretreated intravenously with the nitric oxide donor glyceryl trinitrate (GTN, 250 μg/kg) the mechanically evoked activity decreased after injection of CGRP and increased after injection of olcegepant. In conclusion, the activity of spinal trigeminal neurons with meningeal afferent input is normally not controlled by CGRP receptor activation or inhibition in the trigeminal ganglion. CGRP receptors in the trigeminal ganglion may influence neuronal activity evoked by mechanical stimulation of meningeal afferents only after pretreatment with GTN. Since it has previously been shown that olcegepant applied to the cranial dura mater is ineffective, trigeminal activity driven by meningeal afferent input is more likely to be controlled by CGRP receptors located centrally to the trigeminal ganglion.

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Release of Histamine from Dural Mast Cells by Substance P and Calcitonin Gene-Related Peptide

Cephalalgia ◽

10.1046/j.1468-2982.1997.1703166.x ◽

1997 ◽

Vol 17 (3) ◽

pp. 166-174 ◽

Cited By ~ 133

Author(s):

A Ottosson ◽

L Edvinsson

Keyword(s):

Mast Cells ◽

Substance P ◽

Histamine Release ◽

Dura Mater ◽

Neurogenic Inflammation ◽

Calcitonin Gene Related Peptide ◽

Sampling Procedure ◽

Related Peptide ◽

Calcitonin Gene ◽

Peritoneal Mast Cells

The aim of the present study was to examine if the neuropeptides substance P (SP), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) can stimulate histamine release from mast cells in the dura mater and thereby play a role in cranial vasoregulation and local neurogenic inflammation. Dura mater mast cells were compared with peritoneal mast cells in the rat. Histamine was released from dura mater mast cells by compound 48/80, SP and CGRP but from peritoneal mast cells only by compound 8/80 and SP. NPY and VIP released quite small amounts of histamine from dural mast cells. The release on SP and CGRP from rat dura mater mast cells was blocked by the receptor antagonists FK888 and CGRP8-37 respectively, suggesting receptor mediated release mechanisms. None of the stimuli released histamine from human or porcine dural mast cells, possibly because the sampling procedure injures and incapacitates the cells.

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

Cephalalgia ◽

10.1177/0333102420929027 ◽

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.

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Effect of subarachnoid haemorrhage on trigeminovascular calcitonin-gene-related peptide and substance P of the rat dura mater versus cerebral vasculature

Acta Neurochirurgica ◽

10.1007/bf01808556 ◽

1994 ◽

Vol 127 (1-2) ◽

pp. 103-111 ◽

Cited By ~ 1

Author(s):

A. G. Arand ◽

M. Zuccarello ◽

B. A. Skidmore ◽

R. J. Berger ◽

J. T. Keller

Keyword(s):

Substance P ◽

Subarachnoid Haemorrhage ◽

Dura Mater ◽

Calcitonin Gene Related Peptide ◽

Cerebral Vasculature ◽

Related Peptide ◽

Calcitonin Gene

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Increase in Meningeal Blood Flow by Nitric Oxide - Interaction with Calcitonin Gene-Related Peptide Receptor and Prostaglandin Synthesis Inhibition

Cephalalgia ◽

10.1046/j.1468-2982.2002.00356.x ◽

2002 ◽

Vol 22 (3) ◽

pp. 233-241 ◽

Cited By ~ 34

Author(s):

T Strecker ◽

M Dux ◽

K Messlinger

Keyword(s):

Nitric Oxide ◽

Blood Flow ◽

Dura Mater ◽

Calcitonin Gene Related Peptide ◽

Cyclooxygenase Inhibitors ◽

No Production ◽

Doppler Flowmetry ◽

Related Peptide ◽

Calcitonin Gene ◽

Meningeal Blood Flow

This study addresses possible interactions of the vasodilators nitric oxide (NO), calcitonin gene-related peptide (CGRP) and prostaglandins, which may be implicated in the generation of vascular headaches. Local application of the NO donator diethylamine-NONOate (NONOate) to the exposed dura mater encephali of the rat caused dose-dependent increases in meningeal blood flow recorded by laser Doppler flowmetry. Pre-application of the CGRP receptor antagonist CGRP8-37 significantly attenuated the evoked blood flow increases, while the cyclooxygenase inhibitors acetylsalicylic acid and metamizol were only marginally effective. Stimulation of rat dura mater with NONOate in vitro caused increases in CGRP release. NADPH- diaphorase activity indicating NO production was restricted to the endothelium of dural arterial vessels. We conclude that increases in meningeal blood flow caused by NO depend partly on the release and vasodilatory action of CGRP from dural afferents, while prostaglandins are not significantly involved.

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Interaction of Calcitonin Gene-Related Peptide, Nitric Oxide and Histamine Release in Neurogenic Blood Flow and Afferent Activation in The Rat Cranial Dura Mater

Cephalalgia ◽

10.1111/j.1468-2982.2007.01321.x ◽

2007 ◽

Vol 27 (6) ◽

pp. 481-491 ◽

Cited By ~ 45

Author(s):

N Schwenger ◽

M Dux ◽

R de Col ◽

R Carr ◽

K Messlinger

Keyword(s):

Nitric Oxide ◽

Blood Flow ◽

Histamine Release ◽

Dura Mater ◽

Calcitonin Gene Related Peptide ◽

Receptor Blockade ◽

Cgrp Receptor ◽

Related Peptide ◽

Calcitonin Gene

Calcitonin gene-related peptide (CGRP), nitric oxide (NO) and histamine are implicated in primary headaches but their role in vascular and nociceptive events in the dura mater is not well described. In an in vitro preparation of the hemisected rat skull, CGRP and histamine release from the cranial dura was measured using enzyme-linked immunoassays. While the NO donator NONOate (10-4 M) was without effect, CGRP (10-5 M) induced considerable histamine release from the rat cranial dura, which was blocked by the CGRP receptor antagonist CGRP8-37 (10-5 M). Conversely, histamine (10-4 M) did not stimulate CGRP release. In vitro recordings from single rat meningeal afferents showed that only one of 12 mechanically identified units but several mechanically insensitive units responded to histamine (up to 10-5 M). Increases in meningeal blood flow after histamine application (10-4 M) to the rat cranial dura remained unchanged during CGRP receptor blockade with CGRP8-37, inhibition of NO synthesis with L-NAME (20 mg/kg i.v.) and H3 receptor blockade with thioperamide (10-4 M). We conclude that histamine produces direct vasodilatation and activates a subset of largely non-mechanically sensitive, non-CGRP containing afferents in the rat meninges. Histamine is released from meningeal mast cells which are stimulated by CGRP. Similar mechanisms may be involved in the pathogenesis of headaches.

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Calcitonin gene related peptide released from dural nerve fibers mediates increase of meningeal blood flow in the rat

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y95-143 ◽

1995 ◽

Vol 73 (7) ◽

pp. 1020-1024 ◽

Cited By ~ 52

Author(s):

K. Meßlinger ◽

U. Hanesch ◽

M. Kurosawa ◽

M. Pawlak ◽

R. F. Schmidt

Keyword(s):

Blood Flow ◽

Dura Mater ◽

Calcitonin Gene Related Peptide ◽

Nerve Fibers ◽

Dura Mater Encephali ◽

Related Peptide ◽

Calcitonin Gene ◽

Meningeal Blood Flow ◽

Meningeal Artery ◽

Stimulation Of

The parietal dura mater encephali of the rat was shown by immunohistochemistry to be densely innervated by calcitonin gene related peptide (CGRP) immunoreactive nerve fibers spreading around the medial meningeal artery and its branches. Electrical stimulation of the dural surface (10–20 V, 5–10 Hz, 10–30 min) caused a depletion of CGRP-immunopositive fibers, suggesting a release of CGRP. The dural blood flow around branches of the medial meningeal artery was also monitored with a laser Doppler flowmeter. Short periods (30 s) of electrical stimulation with parameters that presumably released CGRP from nerve fibers caused a repeatable and constant increase of the blood flow for 1–2 min. This evoked increase could dose dependently be inhibited by topical application of the CGRP antagonist hCGRP8–37. Accordingly, administration of hCGRP increased the basal blood flow. We conclude that stimulation of trigeminal afferents innervating the dura mater releases CGRP from peptidergic afferent terminals, thereby causing vasodilatation and increasing the meningeal blood flow, an important element of neurogenic inflammation.Key words: dura mater encephali, afferent nerve fibers, calcitonin gene related peptide, immunohistochemistry, laser Doppler flowmetry.

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ATP sensitive potassium (KATP) channel inhibition: A promising new drug target for migraine

Cephalalgia ◽

10.1177/0333102420925513 ◽

2020 ◽

Vol 40 (7) ◽

pp. 650-664

Author(s):

Sarah L Christensen ◽

Gordon Munro ◽

Steffen Petersen ◽

Anmool Shabir ◽

Inger Jansen-Olesen ◽

...

Keyword(s):

Potassium Channel ◽

Glyceryl Trinitrate ◽

Trigeminal Ganglion ◽

Dura Mater ◽

Calcitonin Gene Related Peptide ◽

Potassium Channel Opener ◽

Related Peptide ◽

Channel Opener ◽

Channel Inhibition ◽

Calcitonin Gene

Background Recently, the adenosine triphosphate (ATP) sensitive potassium channel opener levcromakalim was shown to induce migraine attacks with a far higher incidence than any previous provoking agent such as calcitonin gene-related peptide. Here, we show efficacy of ATP sensitive potassium channel inhibitors in two validated rodent models of migraine. Methods In female spontaneous trigeminal allodynic rats, the sensitivity of the frontal region of the head was tested by an electronic von Frey filament device. In mice, cutaneous hypersensitivity was induced by repeated glyceryl trinitrate or levcromakalim injections over nine days, as measured with von Frey filaments in the hindpaw. Release of calcitonin gene-related peptide from dura mater and trigeminal ganglion was studied ex vivo. Results The ATP sensitive potassium channel inhibitor glibenclamide attenuated the spontaneous cephalic hypersensitivity in spontaneous trigeminal allodynic rats and glyceryl trinitrate-induced hypersensitivity of the hindpaw in mice. It also inhibited CGRP release from dura mater and the trigeminal ganglion isolated from spontaneous trigeminal allodynic rats. The hypersensitivity was also diminished by the structurally different ATP sensitive potassium channel inhibitor gliquidone. Mice injected with the ATP sensitive potassium channel opener levcromakalim developed a progressive hypersensitivity that was completely blocked by glibenclamide, confirming target engagement. Conclusion The results suggest that ATP sensitive potassium channel inhibitors could be novel and highly effective drugs in the treatment of migraine.

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