Calcitonin receptor-like receptor (CLR), receptor activity-modifying protein 1 (RAMP1), and calcitonin gene-related peptide (CGRP) immunoreactivity in the rat trigeminovascular system: differences between peripheral and central CGRP receptor distribution.

Calcitonin gene-related peptide (CGRP) is a key mediator in primary headaches including migraine. Animal models of meningeal nociception demonstrate both peripheral and central CGRP effects; however, the target structures remain unclear. To study the distribution of CGRP receptors in the rat trigeminovascular system we used antibodies recognizing two components of the CGRP receptor, the calcitonin receptor-like receptor (CLR) and the receptor activity-modifying protein 1 (RAMP1). In the cranial dura mater, CLR and RAMP1 immunoreactivity (-ir) was found within arterial blood vessels, mononuclear cells, and Schwann cells, but not sensory axons. In the trigeminal ganglion, besides Schwann and satellite cells, CLR- and RAMP1-ir was found in subpopulations of CGRP-ir neurons where colocalization of CGRP- and RAMP1-ir was very rare ( approximately 0.6%). CLR- and RAMP1-ir was present on central, but not peripheral, axons. In the spinal trigeminal nucleus, CLR- and RAMP1-ir was localized to "glomerular structures," partly colocalized with CGRP-ir. However, CLR- and RAMP1-ir was lacking in central glia and neuronal cell bodies. We conclude that CGRP receptors are associated with structural targets of known CGRP effects (vasodilation, mast cell degranulation) and targets of unknown function (Schwann cells). In the spinal trigeminal nucleus, CGRP receptors are probably located on neuronal processes, including primary afferent endings, suggesting involvement in presynaptic regulation of nociceptive transmission. Thus, in the trigeminovascular system CGRP receptor localization suggests multiple targets for CGRP in the pathogenesis of primary headaches.

Glutamatergic transmission in the trigeminal nucleus assessed with local blood flow.

Stimulation of the superior sagittal sinus in humans is pain-producing and in experimental animals leads to excitation of neurons in the caudal trigeminal nucleus and dorsal horns of the C(1/)C(2) cervical spinal cord: the trigeminocervical complex. Neuronal excitation is generally associated with an increase in local blood flow due to flow/metabolism coupling and we have used local blood flow in the trigeminocervical complex to examine the role of N-methyl-D-aspartate (NMDA)-mediated transmission in these neurons. Cats were anaesthetised with alpha-chloralose (60 mg/kg, ip; supplements 20 mg/kg iv) after surgical preparation under halothane (0.5-3%). Animals were paralysed with gallamine triethiodide to prevent possible movement artefact distorting the laser Doppler signals. The superior sagittal sinus was isolated for electrical stimulation (150 V; 250 microsec duration; 0.5, 1, 2, 5, 10 and 20 Hz) and the dorsal surface of the spinal cord exposed at the C(2) level. Blood flow was recorded from the region over the trigeminocervical complex by careful placement of a laser Doppler flow probe. Flow was recorded continuously by an online collection programme and NMDA-mediated transmission modulated by intravenous administration of MK-801 (0.4, 1 and 4 mg/kg, iv) at the stimulation frequency of 5 Hz. Stimulation of the superior sagittal sinus produced a stimulus-locked, frequency-dependent increase in blood flow in the region of the trigeminocervical complex. The mean maximum response was 39+/-4% at 20 Hz. MK-801 had no effect on the resting flow signal but markedly attenuated the SSS-evoked response in a dose-dependent manner. The mean maximum response after 4 mg/kg MK-801 was 13+/-2%. NMDA-mediated transmission is likely to be involved in nociceptive trigeminovascular transmission within the trigeminocervical complex and offers a possible target for both acute and preventative treatment of migraine.

Attenuation of Fos-like immunoreactivity in the trigeminal nucleus caudalis following trigeminovascular activation in the anaesthetised guinea-pig.

The present study has examined the involvement of sensory neurotransmitters in activating neurones in the trigeminal nucleus caudalis following stimulation of the trigeminovascular system in anaesthetised guinea-pigs. Electrical stimulation of the right trigeminal ganglion produced a unilateral expression of Fos-like immunoreactivity (Fos-LI) in the trigeminal nucleus caudalis. The tachykinin NK1 receptor antagonist, GR205171 (100 micrograms/kg i.v.) and the N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801 (1 mg/kg i.v.) each inhibited expression of Fos-LI following electrical stimulation. The calcitonin gene-related peptide (CGRP) receptor antagonist, CGRP8-37 (1.3 mg/kg i.v.), administered following rostral intracarotid infusion of mannitol to disrupt the blood-brain barrier, tended to reduce Fos-LI evoked by electrical stimulation, although this failed to reach statistical significance. Capsaicin (10 nmol in 0.1 ml), administered intracisternally, produced a bilateral expression of Fos-LI in the trigeminal nucleus caudalis. This expression was unaffected by the peripherally acting NK1 receptor antagonist, GR82334 (0.2 mg/kg i.v.) or CGRP8-37 (1.3 mg/kg i.v.). The centrally penetrant NK1 receptor antagonist, GR205171 (100 micrograms/kg i.v.), inhibited significantly Fos-LI evoked by intracisternal capsaicin administration. It is concluded that the sensory neurotransmitters, substance P and glutamate are released centrally following activation of the trigeminovascular system and that each may be involved in activation of cells in the trigeminal nucleus caudalis.

Trigeminal ganglion elicited increases in nucleus trigeminal caudalis blood flow: a novel migraine model.

We developed an assay which predicts the antimigraine efficacy of sumatriptan. Our assay is based on two assumptions: (1) electrical stimulation of the trigeminal ganglion mimics the neurogenic inflammatory process and (2) stimulation-induced increases in n. trigeminal caudalis blood flow reflect activation of a large population of neurons. Briefly, the trigeminal ganglion was electrically stimulated for 30 s periods at 1 and 10 Hz before and after administration of saline or the antimigraine compound sumatriptan in chloralose-anesthetized cats. Sumatriptan blunted the increase in blood flow following stimulation of the trigeminal ganglion. These data suggest that the n. trigeminal caudalis blood flow model may be useful in identifying antimigraine compounds.

Changes in nucleus tractus spinalis nervi trigemini after temporary occlusion of aorta thoracica in dog.

In a group of seven dogs the thoracic aorta was occluded by means of tourniquet for 30 minutes. After the release of aorta the animals were left living for 6-7 days. The spinal cord and brain stem were treated by Nauta-Gygax method after sacrificing the animals, then the preterminal and terminal degenerations of some ascending spinal systems were analysed. Distinct degenerations in nucleus tractus spinalis nervi trigemini were detected. They originated in lower thoracic and lumbosacral segments of spinal cord. Degenerations were localised in the caudal part of nuclei of trigeminal complex. It looks like the above mentioned projections terminating in nucleus tractus spinalis nervi trigemini are related to tractus spinocerebellaris posterior.

Effects of 7-ethoxycarbonyl-6,8-dimethyl-4-hydroxymethyl-1(2H)-phthalazinone (EG626) on the spinal trigeminal nucleus, ventral posteromedial nucleus, and sensory cortex.

Effects of 7-ethoxycarbonyl-6,8-dimethyl-4-hydroxymethyl-1 (2H)-phthalazinone (EG626) on the spinal trigeminal nucleus (STN), ventral posteromedial nucleus (VPM), and sensory cortex were examined in cats anesthetized with alpha-chloralose in comparison with the effects of morphine. EG626 produced a dose-dependent inhibition of the polysynaptic components of the cortical field potentials upon VPM stimulation and either facilitatory or inhibitory effects on the polysynaptic components of the VPM field potential upon stimulation of the medial lemniscus, while the drug failed to affect the STN field potential with trigeminal nerve stimulation. Morphine inhibited the postsynaptic components of the STN field potentials and to a lesser extent, the polysynaptic components of the cortical field potential; and the effects of morphine on the VPM field potential were similar to those seen with EG626. Pretreatment of the animal with naloxone antagonized the facilitatory effect on the VPM field potentials produced by morphine, but not those by EG626. Morphine and EG626 induced either a prolonged increase in the blood flow or transient increase followed by a decrease in the blood flow in the VPM. These results suggest that EG626 may impair the polysynaptic transmission and/or neuron excitability in the sensory cortex and the VPM at least partly due to the change in blood flow there as does morphine. Unlike morphine, however, EG626 did not produce any obvious effect on the STN.