Spontaneous activity in C-fibres after partial damage to the saphenous nerve in mice: Effects of retigabine.

BACKGROUND: Spontaneous pain is the most devastating positive symptom in neuropathic pain patients. Recent data show a direct relationship between spontaneous discharges in C-fibres and spontaneous pain in neuropathic patients. Unfortunately, to date there is a lack of experimental animal models for drug testing. METHODS: We recorded afferent fibres from a new experimental model in vitro. The preparation contains a neuroma formed in a peripheral branch of the saphenous nerve together with the undamaged branches, which maintain intact terminals in a skin flap. RESULTS: Fibres with stable rates of ectopic spontaneous discharges were found among axotomized (5 A- and 18 C-fibres, mean discharge 0.48 ± 0.08 Hz) and 'putative intact' fibres (12 C-fibres, mean discharge 0.28 ± 0.08 Hz). A proportion (~9%) of axotomized fibres had mechanical receptive fields in the skin far beyond the site of injury. Collision experiments demonstrated that action potentials evoked from neuroma and skin travelled by the same fibre, indicating functional cross-talk between neuromatose and putative intact fibres. Retigabine, the specific Kv7 channel opener, depressed spontaneous discharges by 70% in 15/18 units tested. In contrast, responses to mechanical stimulation of the skin were unaltered by retigabine. CONCLUSIONS: Partial damage to a peripheral nerve may increase the incidence of spontaneous activity in C-fibres. Retigabine reduced spontaneous activity but not stimulus-evoked activity, suggesting an important role for ion channels in the control of spontaneous pain and demonstrating the utility of the model for the testing of compounds in clinically relevant variables. WHAT DOES THIS STUDY ADD?: Our in vitro experimental model of peripheral neuropathy allows for pharmacological characterization of spontaneously active fibres. Using this model, we show that retigabine inhibits aberrant spontaneous discharges without altering physiological responses in primary afferents.

Effects of centrally acting analgesics on spinal segmental reflexes and wind-up.

BACKGROUND: The spinal cord is a prime site of action for analgesia. Here we characterize the effects of established analgesics on segmental spinal reflexes. The aim of the study was to look for the pattern of action or signature of analgesic effects on these reflexes. METHODS: We used a spinal cord in vitro preparation of neonate mice to record ventral root responses to dorsal root stimulation. Pregabalin, clonidine, morphine and duloxetine and an experimental sigma-1 receptor antagonist (S1RA) were applied to the preparation in a cumulative concentration protocol. Drug effects on the wind-up produced by repetitive stimulation of C-fibres and on responses to single A- and C-fibre intensity stimuli were analysed. RESULTS: All compounds produced a concentration-dependent inhibition of total spikes elicited by repetitive stimulation. Concentrations producing ∼50% reduction in this parameter were (in µM) clonidine (0.01), morphine (0.1), pregabalin (1), duloxetine (10) and S1RA (30). At these concentrations clonidine, pregabalin and S1RA had significant effects on the wind-up index and little depressant effects on responses to single stimuli. Morphine and duloxetine did not depress wind-up index and showed large effects on responses to single stimuli. None of the compounds had strong effects on the amplitude of the non-nociceptive monosynaptic reflex. CONCLUSIONS: morphine and duloxetine had general depressant effects on spinal reflexes, whereas the effects of clonidine, pregabalin and S1RA appeared to be restricted to signals originated by strong repetitive activation of C-fibres. Results are discussed in the context of reported behavioural effects of the compounds studied.

GABAA α5 subunit-containing receptors do not contribute to reversal of inflammatory-induced spinal sensitization as indicated by the unique selectivity profile of the GABAA receptor allosteric modulator NS16085.

GABAA receptor positive allosteric modulators (PAMs) mediate robust analgesia in animal models of pathological pain. Restoration of diminished spinal GABAA-α2 and -α3 subunit-containing receptor function is a principal contributor to this analgesia, albeit involvement of GABAA-α5-receptors has not been excluded. Thus, we compared NS11394 and TPA023 (PAMs with selectivity/efficacy at GABAA-α2/α3/α5 receptors) with TP003 (a reportedly GABAA-α3 selective PAM) against spinal sensitization. However, in-house electrophysiology studies designed to confirm the selectivity of TPA023 and TP003 for human GABAA receptors did not corroborate published data, with TP003 displaying considerable GABAA-α5 receptor efficacy. Therefore, we identified a novel PAM, NS16085, which possesses negligible efficacy at GABAA-α5 receptors, but with GABAA-α2/α3 efficacy equivalent to NS11394. At the GABAA-α1 receptor the compound gives low level of negative modulation further separating it from the other compounds. Rat pups with carrageenan-induced hindpaw inflammatory hyperalgesia were used to make ex vivo spinal dorsal root-evoked ventral root recordings. Some spontaneous activity and large numbers of spikes to repetitive stimulation of dorsal roots at C-fibre intensity, indicative of wind-up and sensitization were observed. Equimolar concentrations of NS11394, TP003 and NS16085 all attenuated wind-up to a similar degree; TPA023 was clearly less effective. In adult rats, NS16085 (3-30 mg/kg, p.o.) dose-dependently reduced formalin-induced hindpaw flinching with efficacy comparable to NS11394. Thus, potentiation of GABAA-α2 and-α3 receptors is sufficient to depress spinal sensitization and mediate analgesia after inflammatory injury. Positive modulation at GABAA-α5-receptors is apparently dispensable for this process, an important consideration given the role of this receptor subtype in cognitive function.

Inflammation-induced hyperalgesia and spinal microglia reactivity in neonatal rats.

BACKGROUND: Peripheral inflammation and nerve injury evoke pain behaviours in adult rodents mediated by sensitization, a process that involves the activation of microglia in the spinal cord. In neonates, however, peripheral inflammation, but not nerve injury, induces a lasting hyperalgesia. It is known that microglia does not activate after nerve injury in young pups; however, changes in microglia associated with inflammation in neonatal animals have not been studied. METHODS: Inflammation was induced by unilateral intraplantar injection of carrageenan, complete Freund's adjuvant or zymosan in 10-day-old rats. Rats were tested for mechanical sensitivity in response to punctuate stimulation of the dorsal surface of the hind paw using calibrated von Frey filaments. Immunohistochemistry was used to detect changes in size and density of microglial cells using the specific marker Iba-1. The effects of minocycline applications (120 µg, i.t.) on spinal microglia and behaviour induced by zymosan inflammation were studied. RESULTS: Hind paw inflammation in young P10 rats, with either of the agents used, produced an immediate hyperalgesia, which lasted more than 7 days. A concomitant and significant increase in cell size and density in Iba-1-positive cells was observed in the spinal dorsal horn. These morphological changes in spinal microglia were observed as early as 1-h post-inflammation. Intrathecal and systemic administration of minocycline blocked the hyperalgesia and the changes in spinal microglia produced by zymosan. CONCLUSIONS: Results suggest a key role for spinal microglia activation in the development of hyperalgesia following inflammation in neonatal animals.

[Utility of diffusion-weighted magnetic resonance imaging in severe focal traumatic brain injuries]. / Utilidad de la resonancia magnética potenciada en difusión en pacientes con lesiones focales por traumatismo craneoencefálico grave.

OBJECTIVE: To describe the apparent diffusion coefficient (ADC) in a series of severe traumatic brain injuries, their clinical and outcome features, and possible implications. DESIGN: A descriptive, observational case-series study was carried out. PATIENTS AND INTERVENTIONS: Patients with severe traumatic brain injuries (TBIs) admitted to the ICU were subjected to MRI study using a 1.5 T scanner. Diffusion-weighted images (DWMR) were obtained using the following echo-planar pulse sequence: TR 10000 ms, TE 126.9 ms, with b values 1000 s/mm2 in the three spatial dimensions. Combining the three sets of images, an isotropic image conforming a map of the mean ADCs was obtained. RESULTS: DWMR was performed in 23 patients with severe TBI admitted to the ICU between 2001 and 2004. In the MR images we selected 26 regions of interest (ROIs) where ADC was recorded. We observed a clear increase in diffusion in non-treated space-occupying lesions versus other types of injuries and the normal values. A poorer outcome was recorded in patients with lower ADC values. CONCLUSIONS: Mean ADC in the lesions was greater than the normal values and greater in contusions than in other types of injuries, as an expression of extracellular edema. ADCs were decreased in patients with a poor outcome, suggesting an association between ischemia and the patient prognosis.

Pharmacological properties of S1RA, a new sigma-1 receptor antagonist that inhibits neuropathic pain and activity-induced spinal sensitization.

BACKGROUND AND PURPOSE: The sigma-1 (σ(1) ) receptor is a ligand-regulated molecular chaperone that has been involved in pain, but there is limited understanding of the actions associated with its pharmacological modulation. Indeed, the selectivity and pharmacological properties of σ(1) receptor ligands used as pharmacological tools are unclear and the demonstration that σ(1) receptor antagonists have efficacy in reversing central sensitization-related pain sensitivity is still missing. EXPERIMENTAL APPROACH: The pharmacological properties of a novel σ(1) receptor antagonist (S1RA) were first characterized. S1RA was then used to investigate the effect of pharmacological antagonism of σ(1) receptors on in vivo nociception in sensitizing conditions and on in vitro spinal cord sensitization in mice. Drug levels and autoradiographic, ex vivo binding for σ(1) receptor occupancy were measured to substantiate behavioural data. KEY RESULTS: Formalin-induced nociception (both phases), capsaicin-induced mechanical hypersensitivity and sciatic nerve injury-induced mechanical and thermal hypersensitivity were dose-dependently inhibited by systemic administration of S1RA. Occupancy of σ(1) receptors in the CNS was significantly correlated with the antinociceptive effects. No pharmacodynamic tolerance to the antiallodynic and antihyperalgesic effect developed following repeated administration of S1RA to nerve-injured mice. As a mechanistic correlate, electrophysiological recordings demonstrated that pharmacological antagonism of σ(1) receptors attenuated the wind-up responses in spinal cords sensitized by repetitive nociceptive stimulation. CONCLUSIONS AND IMPLICATIONS: These findings contribute to evidence identifying the σ(1) receptor as a modulator of activity-induced spinal sensitization and pain hypersensitivity, and suggest σ(1) receptor antagonists as potential novel treatments for neuropathic pain.

Comparison of the novel subtype-selective GABAA receptor-positive allosteric modulator NS11394 [3'-[5-(1-hydroxy-1-methyl-ethyl)-benzoimidazol-1-yl]-biphenyl-2-carbonitrile] with diazepam, zolpidem, bretazenil, and gaboxadol in rat models of inflammatory and neuropathic pain.

Spinal administration of GABA(A) receptor modulators, such as the benzodiazepine drug diazepam, partially alleviates neuropathic hypersensitivity that manifests as spontaneous pain, allodynia, and hyperalgesia. However, benzodiazepines are hindered by sedative impairments and other side effect issues occurring mainly as a consequence of binding to GABA(A) receptors containing the alpha(1) subunit. Here, we report on the novel subtype-selective GABA(A) receptor-positive modulator NS11394 [3'-[5-(1-hydroxy-1-methyl-ethyl)-benzoimidazol-1-yl]-biphenyl-2-carbonitrile], which possesses a functional efficacy selectivity profile of alpha(5) > alpha(3) > alpha(2) > alpha(1) at GABA(A) alpha subunit-containing receptors. Oral administration of NS11394 (1-30 mg/kg) to rats attenuated spontaneous nociceptive behaviors in response to hindpaw injection of formalin and capsaicin, effects that were blocked by the benzodiazepine site antagonist flumazenil. Ongoing inflammatory nociception, observed as hindpaw weight-bearing deficits after Freund's adjuvant injection, was also completely reversed by NS11394. Likewise, hindpaw mechanical allodynia was fully reversed by NS11394 in two rat models of peripheral neuropathic pain. Importantly, NS11394-mediated antinociception occurred at doses 20 to 40-fold lower than those inducing minor sedative or ataxic impairments. In contrast, putative antinociception associated with administration of either diazepam, zolpidem, or gaboxadol only occurred at doses producing intolerable side effects, whereas bretazenil was completely inactive despite minor influences on motoric function. In electrophysiological studies, NS11394 selectively attenuated spinal nociceptive reflexes and C-fiber-mediated wind-up in vitro pointing to involvement of a spinal site of action. The robust therapeutic window seen with NS11394 in animals suggests that compounds with this in vitro selectivity profile could have potential benefit in clinical treatment of pain in humans.

Simultaneous assessment of the effects of L-type current modulators on sensory and motor pathways of the mouse spinal cord in vitro.

The effects of modulators of L-type currents in the processing of nociceptive stimuli across sensory and motor circuits were studied using an in vitro preparation of the young mouse spinal cord. Responses to repetitive C-fibre intensity stimuli delivered to a lumbar dorsal root were simultaneously recorded from motor axons in the corresponding ventral root and from putative sensory axons in the anterolateral pathway. L-current antagonists verapamil, diltiazem and nimodipine as well as the agonist Bay K8644 were superfused at a range of concentrations and their effects on responses to afferent stimulation were assessed. All antagonists produced a concentration-dependent depression of transmission across sensory and motor pathways by inhibiting sustained firing and wind-up. All antagonists showed concentration-dependent depression of evoked firing in anterolateral fibres with LogIC50 of -4.2 for verapamil, -4.1 for diltiazem and -4.9 for nimodipine. Applied at high concentrations (>or=100 microM) verapamil and diltiazem produced almost complete blockade of the ascending signals whereas nimodipine produced only partial depression. The effects of the antagonists on motor pathways were significantly greater and the LogIC50 decreased to -5 for verapamil, to -4.9 for diltiazem and to -5.3 for nimodipine. Bay K8644 applied at 2 microM produced only a slight potentiation of responses in anterolateral axons and a very large and long-lasting potentiation of responses from motor neurons. We conclude that mice motor pathways are more sensitive to L-type current modulators than the anterolateral pathway and that analgesic effects reported for some L-type antagonists may be due to a mixture of selective and non-selective effects of these agents on sensory neurones.

Retigabine-induced population primary afferent hyperpolarisation in vitro.

Retigabine is a compound of potential interest in analgesia which acts as an M-channel opener to depress neuronal excitability. Here we study the effects of retigabine and its antagonist XE-991 on populations of primary afferents. Experiments were performed using a hemisected spinal cord preparation from rat pups maintained under in vitro conditions. Recording from dorsal roots were performed using tight fitting suction electrodes coupled to AC and DC amplifiers. The adjacent dorsal root was electrically stimulated at regular intervals. The effects of the modulators on basal potential, spontaneous potentials and dorsal root-dorsal root responses were studied. Superfusion of retigabine (10 microM) produced long lasting and robust hyperpolarisation of primary afferents which persisted during superfusion of picrotoxin (20 microM) and tetrodotoxin (0.5 microM). Other effects of retigabine were (1) increase in stimulation threshold; (2) increase in size of responses to suprathreshold stimuli; and (3) increase in amplitude and decrease in frequency of spontaneous dorsal root potentials. Superfusion of XE-991 had little effect on its own but blocked all the effects of retigabine. These results indicate the presence of functional M-currents in central terminals of primary afferents and in the interneurones that mediate dorsal root potentials. The depressant effects of retigabine on the excitability of these structures may contribute to its analgesic effects after pain-inducing treatments.

Effects of M-current modulators on the excitability of immature rat spinal sensory and motor neurones.

M-currents have been shown to control neuronal excitability in a variety of central and peripheral neurones. Here we studied the effects of specific M-current modulators on the excitability of spinal neurones and their response to synaptic activation. Experiments were performed in vitro using the hemisected spinal cord from 7- to 11-day-old rats. Intracellular recordings were obtained from lumbar deep dorsal horn and motor neurones. Neuronal excitability was assessed by applying outward current pulses and synaptic responses were elicited by activation of a lumbar dorsal root. The M-current antagonist 10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone (XE-991) and the agonist retigabine were superfused at 10 microM. Retigabine produced hyperpolarization and a large decrease in the excitability of motor (7/7) and dorsal horn neurones (11/12). The effects of retigabine were fully reversed by XE-991. XE-991 induced depolarization of most neurones tested and a large increase in the excitability of motor neurones (7/7) but only a weak increase in the excitability of a proportion of dorsal horn neurones (4/10). The effects of XE-991 were partly reversed by retigabine. Consistent with their effects on neuronal excitability, retigabine showed a general depressant effect on synaptic transmission, whereas XE-991 showed the opposite tendency to potentiate responses to dorsal root stimulation, particularly in motor neurones. The results show that retigabine can depress spinal excitability and the transmission of nociceptive information. Results also indicate a post-synaptic expression of functional M-currents in most motor neurones and a considerable proportion of deep dorsal horn neurones.

Electrophysiological and pharmacological characterisation of ascending anterolateral axons in the in vitro mouse spinal cord.

Here we describe a procedure to obtain electrophysiological recordings from cut ends of ascending axons of the spinal cord white matter. This procedure involves the use of an in vitro preparation of the neonate mouse spinal cord and suction microelectrodes of about 10 microm tip diameter. Using this procedure, we have recorded from axons of the anterolateral quadrant at the thoracic level which responded to electrical stimulation of a lumbar dorsal root. Using adequate stimulation parameters, we have identified axons responding exclusively to activation of C-fibres and axons responding to activation of A- and C-fibres. These axons are likely to originate at dorsal horn second order sensory neurons of different functional types. Simultaneous recordings from a lumbar ventral root and from ascending axons were used to assess the effects of noradrenaline and sevoflurane on motor and sensory pathways. The results obtained show that noradrenaline can potentiate responses of motor neurons to dorsal root stimulation while depressing responses of ascending axons to the same stimulus. In contrast both motor and sensory pathways were inhibited by sevoflurane. We believe that the procedures developed here may be of great interest to assess spinal nociceptive and antinociceptive mechanisms in vitro.

Effects of propofol and sevoflurane on the excitability of rat spinal motoneurones and nociceptive reflexes in vitro.

BACKGROUND: Spinal actions of halogenated ethers are widely recognized, whereas spinal actions of intravenous anaesthetics like propofol are less clear. The aim of this study was to compare the spinal effects of propofol and sevoflurane. METHODS: We used an isolated spinal cord in vitro preparation from rat pups and superfused the anaesthetics at known concentrations. Responses of motoneurones to single and repetitive C-fibre intensity stimulation (trains of 20 stimuli at 1 Hz) of a lumbar dorsal root were recorded from the corresponding ventral root via a suction electrode. RESULTS: Stimulation trains produced a wind-up of action potentials in motoneurones. Both propofol and sevoflurane produced a significant concentration-dependent depression of the evoked wind-up, although at clinically relevant concentrations sevoflurane exhibited a larger intrinsic efficacy. Applied at anaesthetic concentrations, sevoflurane 250 micro M abolished action potentials whereas propofol 1 micro M only produced a reduction close to 50%. At these concentrations, sevoflurane produced a large depressant effect on the monosynaptic reflex whereas propofol was ineffective. CONCLUSIONS: Sevoflurane produces large inhibitory effects on nociceptive and non-nociceptive reflexes which are likely to contribute to immobility during surgery. Compared with sevoflurane, propofol appears to have much weaker effects on spinal reflexes such as those recorded in an isolated preparation.

M-current modulators alter rat spinal nociceptive transmission: an electrophysiological study in vitro.

M-currents constitute a unique effector system to control neuronal excitability due to their voltage and ligand sensitivities. Here we have used retigabine, an M-current agonist, and XE-991, an M-current antagonist, to study the possible involvement of these currents in the processing of spinal sensory and motor processing of nociceptive information in normal, untreated rats. Experiments were performed in a hemisected spinal cord preparation from rat pups using extracellular recordings. Responses to activation of nociceptive and non-nociceptive afferent fibres were recorded. M-current modulators were bath applied to the entire cord or applied locally by pressure ejection. Retigabine and XE-991 produced long-lasting and concentration-dependent effects on nociceptive reflexes showing only minor effects on non-nociceptive reflexes. Retigabine depressed responses to repetitive stimulation of the dorsal root recorded from motor neurones and dorsal horn neurones, whereas XE-991 showed the opposite potentiatory effect and reversed effects of retigabine. Local application of the modulators close by motor nuclei produced changes in reflex responses similar to those caused by bath application. These results constitute a clear indication of the existence of functional M-currents in dorsal and ventral horn elements of the mammalian spinal cord where they may serve to regulate early sensory and motor processing of nociceptive information. The weak effect of modulators on non-nociceptive reflexes suggest that M-currents constitute a promising novel target for analgesics.

Characterisation of sevoflurane effects on spinal somato-motor nociceptive and non-nociceptive transmission in neonatal rat spinal cord: an electrophysiological study in vitro.

Sevoflurane is the latest halogenated ether introduced in clinical anaesthesia, and its effects at the spinal level are not fully characterised. The rat hemisected spinal cord preparation was used to test the effects of sevoflurane on spinal nociceptive and non-nociceptive synaptic transmission as well as on excitations produced by application of glutamate-receptor agonists. Sevoflurane was dissolved in artificial cerebrospinal fluid (ACSF) with a specific vaporiser, and its final concentration was assessed with gas chromatography. Sevoflurane reduced the mono-synaptic reflex (EC(50) approximately 219 microM) and the slow components of the dorsal root-ventral root potentials (EC(50) approximately 72 microM) elicited by single dorsal root stimulation as well as the cumulative depolarisation (CD) elicited by repetitive stimulation (EC(50) approximately 98 microM). AMPA- and NMDA-induced depolarisations were also reduced by sevoflurane (respective EC(50)s were 206 and 127 microM). Inhibition of NMDA-induced depolarisation was TTX resistant. However, complete blockade of NMDA receptors with d-AP5 did not prevent further reduction of the CD by sevoflurane. All the effects reported were concentration-dependent and reversible. We conclude that sevoflurane applied at clinically relevant concentrations induces a strong depression of nociceptive and non-nociceptive spinal systems, which may be partly mediated by interfering with excitatory amino acid transmission.

5-HT(1B) but not 5-HT(6) or 5-HT(7) receptors mediate depression of spinal nociceptive reflexes in vitro.

1. The identity of the serotonin (5-HT) receptors modulating the transmission of segmental C-fibre mediated signals was studied using an in vitro preparation of the hemisected spinal cord from rat pups. 2. Responses to trains of stimuli delivered to a lumbar dorsal root were recorded from the corresponding ventral root. The resulting cumulative depolarization (CD) mediated by unmyelinated fibres was quantified in terms of integrated area. The amplitude of the mono-synaptic reflex was also measured. Serotonergic agents were superfused at known concentrations and their effects on the reflexes evaluated. 3. 5-HT had depressant effects on the CD (EC(50) 34 microM). The rank order of potency of agonists for the depression of the CD was 5-carboxamidotryptamine (5-CT)>alpha-methylserotonin (alpha-met-5-HT) approximately 5-HT>42-methylserotonin (2-met-5-HT)approximately 8-OH-DPAT. 4. All the agonists including 2-met-5-HT and 8-OH-DPAT had strong depressant effects on the mono-synaptic reflex with the following order of potency: 5-CT>48-OH-DPAT>4alpha-met-5-HT approximate5-HTapproximate2-met-5-HT. 5. The inhibitory effects of 5-HT, alpha-met-5-HT and 5-CT were attenuated by the non-specific 5-HT antagonist methiothepin (1 microM) and by the 5-HT(1A/1B) antagonist SDZ 21009 (100 nM) but not by the selective 5-HT(1A) antagonist WAY 100135 (1 microM). 6. Other antagonists known to block 5-HT(2), 5-HT(6) and/or 5-HT(7) receptors (ketanserin, RO 04-6790, ritanserin and clozapine) did not change the effect of the agonists. 7. The data suggest an important contribution of 5-HT(1B) receptors to the inhibition of spinal C-fibre mediated nociceptive reflexes but no experimental support was found for the intervention of 5-HT(2), 5-HT(6) or 5-HT(7) receptors in this in vitro model.

Alpha-1A adrenoceptors modulate potentiation of spinal nociceptive pathways in the rat spinal cord in vitro.

The rat hemisected spinal cord preparation was used to assess the role of different adrenoceptor subtypes on the modulation of nociceptive reflexes. These were elicited by trains of high intensity electrical stimuli delivered to a lumbar dorsal root. Responses were recorded from the corresponding ventral root in AC- and DC-amplification modes simultaneously. Superfusion of noradrenaline produced a potentiation of action potential firing (AC channel) as well as a depression of the cumulative depolarisation (DC channel) in responses to repetitive afferent stimulation.Noradrenaline-induced potentiation of firing was mimicked by the alpha1A-adrenoceptor agonist A 61603 and the alpha1-adrenoceptor agonist methoxamine in a reversible and concentration-dependent manner. The order of potency of these agonists was A61603>>noradrenaline>methoxamine. The alpha(1A)-adrenoceptor antagonist 5-methyl-urapidil and the alpha1-adrenoceptor antagonist corynanthyne blocked the excitatory effects of noradrenaline. In contrast, the alpha(1B/D)-adrenoceptor antagonists chloroethylclonidine and BMY 7378 failed to block this effect.Noradrenaline-induced depression of cumulative depolarisation was mimicked by the alpha2-adrenoceptor agonist UK 14,304. In addition, this compound produced inhibition of firing in responses to afferent stimulation. These results show that noradrenaline has bi-directional modulatory effects on nociceptive reflexes and indicate that selective activation of alpha1A- but not alpha1B/D-adrenoceptors mediate potentiation of spinal nociceptive reflexes.

Responses of rat spinal neurons to distension of inflamed colon: role of tachykinin NK2 receptors.

Tachykinin NK2 receptors are implicated in nociception and the control of intestinal motility. Here we examined their involvement in responses of spinal lumbosacral neurons with colon input to distension of normal or inflamed colon in anesthetized rats. The responses of single neurons to colorectal distension (5-80 mmHg), to electrical stimulation of the pelvic nerve (bypassing sensory receptors) and to somatic stimulation were characterized. The effect of cumulative doses of an NK2 receptor antagonist, MEN 11420 (10-1000 microg kg(-1) IV), on responses to these stimuli was tested in control conditions (n=6), or 45 min after intracolonic instillation of acetic acid (n=6). After colonic inflammation, neuronal responses to colorectal distension and pelvic nerve stimulation were significantly greater. MEN 11420 dose-dependently inhibited the enhanced responses to colorectal distension after inflammation (ID50=402+/-14 microg kg(-1)), but had no significant effect on responses to pelvic nerve stimulation or distension of the normal colon, suggesting a peripheral action selective for the inflamed colon. We conclude that MEN 11420 possesses peripheral anti-hyperalgesic effects on neuronal responses to colorectal distension. These results provide a neurophysiological basis for a possible use of tachykinin NK2 receptor antagonists in treating abdominal pain in irritable bowel syndrome patients.

Wind-up of spinal cord neurones and pain sensation: much ado about something?

Wind-up is a frequency-dependent increase in the excitability of spinal cord neurones, evoked by electrical stimulation of afferent C-fibres. Although it has been studied over the past thirty years, there are still uncertainties about its physiological meaning. Glutamate (NMDA) and tachykinin NK1 receptors are required to generate wind-up and therefore a positive modulation between these two receptor types has been suggested by some authors. However, most drugs capable of reducing the excitability of spinal cord neurones, including opioids and NSAIDs, can also reduce or even abolish wind-up. Thus, other theories involving synaptic efficacy, potassium channels, calcium channels, etc. have also been proposed for the generation of this phenomenon. Whatever the mechanisms involved in its generation, wind-up has been interpreted as a system for the amplification in the spinal cord of the nociceptive message that arrives from peripheral nociceptors connected to C-fibres. This probably reflects the physiological system activated in the spinal cord after an intense or persistent barrage of afferent nociceptive impulses. On the other hand, wind-up, central sensitisation and hyperalgesia are not the same phenomena, although they may share common properties. Wind-up can be an important tool to study the processing of nociceptive information in the spinal cord, and the central effects of drugs that modulate the nociceptive system. This paper reviews the physiological and pharmacological data on wind-up of spinal cord neurones, and the perceptual correlates of wind-up in human subjects, in the context of its possible relation to the triggering of hyperalgesic states, and also the multiple factors which contribute to the generation of wind-up.