Effects of superfusion of morphine and enkephalins on the activity of single units in the spinal trigeminal nucleus and cuneate nucleus of cat

The effects of superfusion of morphine, met-enkephalin and D-ala2-met5-enkephalinamide on the spontaneous neural discharge rates of units in the spinal trigeminal nucleus and cuneate nucleus of decerebrate cats were studied. The drugs were superfused onto the dorsum of the exposed surface of the caudal medulla overlying these nuclei. Some of these neurons were identified by their response to innocuous mechanical stimuli delivered to the skin. In the caudal spinal trigeminal nucleus, morphine caused a dose-dependent suppression of the spontaneous discharge rate in the majority of the neurons studied. Endogenous opiate peptide, met-enkephalin or its synthetic analogue, D-ala2-met5-enkephalinamide caused an initial reduction, followed by a rebound of the discharge rate to the control value. These depressant effects of morphine and enkephalins were antagonized by concomitant superfusion of the opiate antagonist naloxone. In the main cuneate nucleus, however, similar doses of morphine, met-enkephalin and D-ala2-met5-enkephalinamide have little if any significant effect on the spontaneous activity of the neurons studied. These results provide electrophysiological evidence for the presence of opiate receptors in the caudal spinal trigeminal nucleus and the relative lack of such receptors in the main cuneate nucleus.     

Effect of migrepin on activity of trigeminal nucleus caudalis neurons

Neurophysiological experiments on anesthetized rats were used to study the effects of various doses (12.5, 25, 37.5 mg/kg, i.v.) of drug composition migrepin (representing a combination of potassium-2,4-dichlorobenzoate, carbamazepine, and caffeine) on background firing of the trigeminal nucleus caudalis neurons and their responses to electrical stimulation of the dura mater. It was found that migrepin produces direct, dose-dependent inhibitory action on functional activity of TNC neurons. The results confirmed anti-migraine properties of the drug but did not exclude the necessity to study its action in clinical trials.     

Effects of phaclofen and the enantiomers of baclofen on cardiovascular responses to intrathecal administration of L- and D-baclofen in the rat.

In a previous study it was found that i.t. administration of L-baclofen decreased arterial pressure and heart rate while D-baclofen differentially increased arterial pressure. The objective of the present study was to determine which of these effects was blocked by prior administration of the GABAB receptor antagonist, phaclofen, and whether the effect of one enantiomer of baclofen could be blocked by prior administration of the other. The decreases in systolic and diastolic arterial pressures and in heart rate produced by i.t. administration of 70 nmol of L-baclofen were unaffected by i.t. administration of 7, 70 or 700 nmol of D-baclofen 10 min prior to administration of L-baclofen, but were blocked by administration of 5 mumol of phaclofen given 3-5 min prior to L-baclofen. On the other hand, the increases in systolic and diastolic arterial pressures induced by i.t. administration of 700 nmol of D-baclofen were blocked by 70 nmol but not by 7 nmol of L-baclofen, as well as by 2.5 mumol of phaclofen; the effect of L-baclofen cannot be attributed to a desensitization of D-baclofen-sensitive receptors as two successive doses of D-baclofen given 7 min apart had quantitatively similar effects. Phaclofen alone increased systolic and diastolic arterial pressures and heart rate. The results are interpreted as indicating that D-baclofen is not an antagonist of L-baclofen in this paradigm; rather, they suggest that L-baclofen reduces the effects of D-baclofen.(ABSTRACT TRUNCATED AT 250 WORDS)     

The action of GABAB antagonists in the trigeminal nucleus of the rat.

The iontophoretic administration of the GABAB antagonists (P-(3-aminopropyl)-P-diethoxymethyl-phosphinic acid (CGP 35348) and 2-hydroxy-saclofen blocked the action of iontophoretically applied L-baclofen on neurons in the trigeminal nucleus of rats, anesthetized with halothane. The substance CGP 35348 appeared to be more potent than 2-hydroxy-saclofen. The iontophoretic administration of GABA resembled L-baclofen in depressing excitatory transmission and facilitating segmental inhibition in the trigeminal nucleus. The depression of excitatory transmission was also blocked by CGP 35348 and the facilitation of segmental inhibition produced by GABA was partially blocked. These observations indicate that CGP 35348 is not only a baclofen antagonist but actually a GABAB receptor antagonist and the baclofen was acting at GABAB receptors in the trigeminal nucleus. The portion of the effect of GABA, not blocked by CGP 35348, was probably mediated by GABAA receptors, since it was previously found that segmental inhibition in the trigeminal nucleus could be modulated by GABAA agonists and antagonists as well.     

Cardiovascular responses to intrathecal administration of L- and D-baclofen in the rat.

D- and L-baclofen were given intrathecally at the T2 spinal level in the anaesthetized rat. D-Baclofen, in doses of 7, 35 and 70 nmol produced graded increases in arterial pressure but heart rate remained unaffected. Responses appeared within 30 s, peaked at 2 min and decayed over the next 5 min. Injection i.v. of 70 nmol of D-baclofen failed to alter arterial pressure or heart rate. In contrast, intrathecal administration of L-baclofen decreased both arterial pressure and heart rate. The amplitude and time course of the effects depended on the dose used; 700 nmol of L-baclofen had stronger and longer effects than those induced by 70 nmol, while 7 nmol had no effect. (I.v. injection of 70 nmol of L-baclofen had similar effects to intrathecal administration but with different time course and amplitude.) When given at the T9 level at doses of 70 nmol, D- and L-baclofen had effects similar to those observed at the second thoracic level. Effects of intrathecal administration of D- and L-baclofen at T2 were prevented by pretreatment with either hexamethonium (10 mg/kg i.v.) or lidocaine (25 microliters of a 1% solution, intrathecally). The results suggest that D- and L-baclofen-sensitive receptors in the spinal cord are involved in regulating sympathetic output in pathways to the vessels and/or to the heart. In addition, our results suggest that D- or L-baclofen may not act via classical GABAB receptors or that two types of GABAB receptor exist in spinal sympathetic pathways.     

Effects of opioids and opioid peptide on the release of substance P-like material induced by tooth pulp stimulation in the trigeminal nucleus caudalis of the rabbit

The superficial layer in subnucleus caudalis of the brain-stem trigeminal sensory nuclear complex (SpVc) in the rabbit was perfused with artificial cerebrospinal fluid using a push-pull perfusion cannula system. Immunoreactive substance P (iSP) and [Met5]enkephalin (iME) released into the perfusates following electrical stimulation of the lower incisor pulp were measured. An increase in the release of iSP and iME lasting for 1 h or more was observed following electrical stimulation with 40 V. The increase in iSP release depended on the intensity of stimulation. Systemic morphine (10 mg/kg i.v.) completely inhibited the stimulus-evoked iSP release and this inhibition was antagonized by pretreatment with naloxone (5 mg/kg i.v.). The stimulus-evoked iSP release was also inhibited by local application of morphine (10(-6) M) or the opioid peptide [D-Ala2,Met5]enkephalinamide (10(-4) M). However, the local application of naloxone (5 X 10(-7) M) only partially antagonized the inhibitory effects of locally applied morphine and the opioid peptide. These results suggest that there is a functional interaction between SP and enkephalin systems in the superficial layer of SpVc for the regulation of dental pain transmission.     

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.     

Antagonizing effects of beta-adrenergic blockers on lucus coeruleus-induced inhibition of trigeminal nucleus neurons.

Effects of alpha- and beta-adrenergic blocking agents applied into the lateral ventricle were studied on the relay neuron in the rostral part of spinal trigeminal nucleus (STN) of cats. Conditioning stimulation of the locus coeruleus (LC) and sensory cortex (SC) inhibited the orthodromic spike generation in STN relay neuron without affecting the antidromic spike, as already reported, and re-confirmed herein. The LC-induced inhibition of orthodromic spike was significantly reduced by intraventricular administration of beta-blockers, MJ 1999 (5 mg) and propranolol (0.5 mg), while the SC-induced inhibition of orthodromic one was not modified by the beta-blockers. The antidromic spike in STN relay neuron per se remained unaffected by these treatments. Intraventricular administration of alpha-blockers such as phentolamine and phenoxybezamine produced no alterations of the LC- and SC-induced inhibition of orthodromic spike. As the beta-blockers produced a selective antagonism, noradrenaline originating in LC probably acts as an inhibitory transmitter on the STN relay neuron and is mediated by beta-receptor.     

Impariment by 6-hydroxydopamine of locus coeruleus-induced monosynaptic potential in the spinal trigeminal nucleus

All experiments were performed on cats immobilized with gallamine. In the rostral part of spinal trigeminal nucleus (STN), single fiber action potential with a consistent and short latency was elicited by electrical stimulation of the locus coeruleus (LC). When 2 mg of 6-hydroxydopamine (6-OHDA) dissolved in 5% ascorbic acid solution was applied into the lateral ventricle, the STN field potential produced by LC stimulation was reduced in amplitude within 20 hr. The STN potential elicited by trigeminal nerve stimulation, however, was unaffected until 24 hr after 6-OHDA. The drug also blocked the inhibitory effect of LC conditioning stimulation on the STN potential elicited by trigeminal nerve stimulation, whereas it did not modify the inhibitory effect of conditioning stimulation of the sensory cortex on the STN potential. Ascorbic acid solution, a solvent, affected neither the STN potential by LC stimulation, nor the inhibitory effect of LC neurons on the STN potential elicited by trigeminal nerve stimulation. These results strongly suggest the existence of noradrenergic fiber from the LC to STN, through which inhibition of the STN neurons is produced.     

Suppressive effects of oxcarbazepine on tooth pulp-evoked potentials recorded at the trigeminal spinal tract nucleus in cats

1. The purpose of the present study was to evaluate the antinociceptive effect of oxcarbazepine, a keto derivitive of carbamazepine (an anticonvulsant), in an animal model. To evoke a nociceptive response, we electrically stimulated the maxillary canine tooth pulp (MCTP) in anaesthetized (allobarbital-urethane), spontaneously breathing cats. 2. The evoked potentials were recorded from the superficial layers of the caudal part of the trigeminal spinal tract nucleus (5ST). We examined a slow component with a large amplitude (the P3 component) in evoked compound potentials; its mean conduction velocity was 1.7 m/s, suggesting a response mediated by C-fibres. 3. To confirm that the P3 component was related to pain sensation, we used morphine, a most efficacious antinociceptive agent, in the present study. The P3 component was significantly suppressed by intravenous administration of morphine (3 mg/kg) and was also suppressed by microinjection of morphine (2 microg) into the recording site of the 5ST. These results suggest that the P3 component is involved in the transmission of nociceptive information. 4. We compared the effect of oxcarbazepine with mexiletine; both are known to block neuronal Na+ channels. Intravenous administration of mexiletine suppressed the P3 component at a dose of 5 mg/kg. Microinjection of mexiletine (10 microg) into the recording site of the 5ST tended to suppress the P3 component, but this effect was not significant. 5. Intravenous administration of oxcarbazepine (1-10 mg/kg) caused a dose-dependent inhibition of the P3 component, which was significantly suppressed at 10 mg/kg oxcarbazepine. Intravenous administration of 10,11-dihydro-10-hydroxy-5H-dibenz[b,f]azepine-5-carboxamide (MHD), a metabolite of oxcarbazepine, at doses of 3-30 mg/kg caused a dose-dependent inhibition of the P3 component. Oxcarbazepine was not available for the microinjection study because it is not water soluble. We used MHD for the microinjection study instead of oxcarbazepine, because MHD can be dissolved in water up to 3 mg/mL. Microinjections of MHD (6 microg) into the recording site of the 5ST suppressed the P3 component. These results indicate that oxcarbazepine has an antinociceptive action.     

A dose-response analysis of the effects of L-baclofen on chronic tinnitus caused by acoustic trauma in rats

Subjective tinnitus is a chronic neurological disorder in which phantom sounds are perceived. Drugs that increase GABAergic neurotransmission in the CNS are sometimes used as a treatment. One such drug is the GABA(B) receptor agonist L-baclofen. The aim of this study was to investigate the effects of L-baclofen on the psychophysical attributes of tinnitus in rats.The effects of 1, 3 or 5 mg/kg L-baclofen (s.c.) on the psychophysical attributes of tinnitus were investigated using a conditioned lick suppression model, following acoustic trauma (a 16 kHz, 110 dB pure tone presented unilaterally for 1 h) in rats. In pre-drug testing, acoustic trauma resulted in a significant increase in the auditory brainstem-evoked response (ABR) threshold in the affected ear (P < 0.008) and a significant decrease in the suppression ratio (SR) compared to sham controls in response to the 20 kHz tones, but not the broadband noise or the 10 kHz tones (P < 0.002). The 3 and 5 mg/kg doses of L-baclofen significantly reversed the frequency-specific decrease in the SR in the acoustic trauma group, indicating that the drug reduced tinnitus. Following washout from the 3 mg/kg dose, but not the 5 mg/kg dose, the significant decrease in the SR for the acoustic trauma group returned, suggesting a return of the tinnitus. These results suggest that L-baclofen should be reconsidered as a drug treatment for tinnitus. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.     

Disturbed microvascular permeability in the eye caused by capsaicin microinjections into the trigeminal caudalis nucleus

Capsaicin microinjection into the trigeminal caudalis nucleus (the central projection area of trigeminal capsaicin-sensitive nerve) increase extravasation of proteins in rat eye. The effect was inhibited by ruthenium red introduction (a capsaicin receptor antagonist) and by blocking the effector functions of capsaicin-sensitive nerve endings. It is suggested that capsaicin stimulation of central terminations of trigeminal capsaicin-sensitive afferents induce an increase in the microvascular permeability of the eye, which is mediated through the effector function of capsaicin-sensitive nerves.     

Effects of afloqualone on vestibular nystagmus and the lateral vestibular nucleus

To clarify the antivertiginous effect of afloqualone, an antispastic drug, we examined its action on the vestibular nervous system in cats. The results suggest that afloqualone inhibits vestibular nystagmus probably due to both inhibition of selective polysynaptic transmission and enhancement of the effects of GABA and glycine in the lateral vestibular nucleus (LVN), and its GABA-enhancing effect is thought to be attributable to the increased sensitivity of GABA receptors of the LVN neuron site.     

NMDA receptor antagonists block cardiovascular responses to intrathecal administration of D-baclofen in the rat.

In previous studies we found that D and L-baclofen have different effects on sympathetic output when administered intrathecally, yet the actions of both enantiomers are blocked by intrathecal administration of phaclofen. The present experiments were done to determine the mechanism by which D-baclofen expresses its effects. In urethane-anaesthetized Sprague-Dawley rats, when D-baclofen was given intrathecally at the T9 spinal level following pretreatment with 2 nmol of the NMDA receptor antagonist, DL-2-amino-5-phosphonovaleric acid (APV), it increased systolic and diastolic arterial pressures (n = 7), as in the previous studies. However, after intrathecal administration of 10 nmol of APV, administration of D-baclofen had no effect on these parameters (n = 7). Intravenous administration of ketamine (7.5 mg/kg), another NMDA receptor antagonist, also blocked the effect of D-baclofen (n = 6) but it had no effect on the pressor responses produced by intrathecal administration of carbachol (27.4 nmol; n = 6). In additional experiments, L-baclofen (70 nmol) had no effect on the increases in heart rate and arterial pressure produced by N-methyl-D-aspartic acid (NMDA) (2 nmol; n = 8). These results indicate that D-baclofen increases arterial pressure via an NMDA receptor-mediated mechanism, perhaps by provoking the release of an endogenous ligand which activates these receptors.     

Differentiation of excitatory amino acid receptors in the rat caudal trigeminal nucleus: a microiontophoretic study

The excitatory amino acid antagonist, D-alpha-aminoadipate, was found to be a potent antagonist of responses to N-methyl-D-aspartate and L-aspartate, but less potent against L-glutamate responses of neurones in the rat caudal trigeminal nucleus. The responses to quisqualate and kainate were relatively unaffected by D-alpha-aminoadipate, but were antagonized by the two antagonists cis-2,3-piperidine dicarboxylate and gamma-D-glutamylglycine. These two antagonists had effectively similar spectra of activity. It is concluded that there is a distinct N-methyl-D-aspartate receptor in the trigeminal nucleus caudalis, and that kainate and quisqualate may act on a different, but common, receptor. Exogenously applied L-glutamate appears to act at both of these receptors, but exogenously applied L-aspartate appears to act almost exclusively on a D-alpha-aminoadipate sensitive site that may be the N-methyl-D-aspartate receptor. The kainate/quisqualate receptor is thought to be involved in the synaptic excitation of neurones in the caudal trigeminal nucleus by non-noxious mechanical stimuli.     

不同剂量甘丙肽拮抗剂在三叉神经主核对甘丙肽外向电流的双向作用

目的　在三叉神经主核 (PrV)上检测不同剂量甘丙肽拮抗剂M 35和M 4 0对甘丙肽外向电流的作用。方法　全细胞膜片钳记录技术。结果　 0 .0 1、0 .0 3及 0 .1μmol·L- 1的M 35 ,使甘丙肽引发的外向电流分别抑制 5 3.1%、 4 0 .4 %及 2 7.2 % ;但 1、3μmol·L- 1M 35使甘丙肽外向电流分别增大16 .2 %及 38.4 %。而给予 0 .0 1及 0 .0 3μmol·L- 1M 4 0时 ,甘丙肽外向电流增大 93.7%及 6 8.4 % ;0 .3、1及 3μmol·L- 1M 4 0使甘丙肽外向电流分别抑制 4 9.2 %、6 9.9%及 87.2 %。结论　低浓度M 35为甘丙肽拮抗剂 ,高浓度M 35有甘丙肽激动剂作用 ;低浓度M 4 0为甘丙肽激动剂 ,高浓度M 4 0为甘丙肽拮抗剂 ,PrV同一神经元上可能存在不同甘丙肽亚型受体     

Non-NMDA glutamate receptors modulate capsaicin induced c-fos expression within trigeminal nucleus caudalis.

1. We examined the effects of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzol[f]quinoxaline-7-sulpho namide (NBQX), the kainate receptor antagonists gamma-(R-)-glutamylaminomethanesulphonic acid (GAMS) and 6,7,8,9-tetrahydro-5-nitro-1H-benz[g]indole-2,3-dione-3-oxime (NS-102), and the group III metabotropic glutamate receptor (mGluR) agonist 2-amino-4-phosphono-S-butanoic acid (L-AP4) on c-fos-like immunoreactivity (c-fos LI) in trigeminal caudalis (Sp5C), lateral reticular (LRt), medullary reticular (Md) and solitary tract (Sol) nuclei, after intracisternal injection of capsaicin in urethane anaesthetized Sprague-Dawley rats. 2. Few c-fos labelled cells were observed within Sp5C in capsaicin-vehicle treated animals. The number of positive c-fos cells increased by 17 fold after intracisternal capsaicin (5 nmol) administration. 3. Pretreatment with CNQX (0.02, 0.1, 0.6, 3 and 15 mg kg-1) or NBQX (0.01, 0.1 and 1 mg kg-1), administered intraperitoneally 15 min before capsaicin, significantly reduced labelled cells within Sp5C by a maximum of 45 and 34%, respectively. The number of c-fox LI cells within LRt, Md and Sol was not affected. Pretreatment with L-AP4 (1, 3 and 10 mg kg-1) decreased the number of Sp5C c-fos LI cells by a maximum of 30%, whereas GAMS (1 and 10 mg kg-1) and NS-102 (1 and 5 mg kg-1) did not show any significant effect. 4. These results suggest that blockade of AMPA receptors, but not kainate receptors, or the activation of group III mGluRs, decrease the response of Sp5C neurons to trigeminovascular activation. Thus, in addition to NMDA receptors, mGluRs and AMPA receptors may modulate cephalic pain and may provide a potential therapeutic target for antimigraine drugs.