Ascending inhibition of nociceptive neurons in the nucleus ventralis posterolateralis following conditioning stimulation of the nucleus raphe magnus

Recordings were made from neurons in the nucleus ventralis posterolateralis (VPL) of urethane-chloralose-anesthetized cats, following both noxious mechanical stimulation of the integument and electrical stimulation of the greater splanchnic nerve (SPL). The effects of stimulating the nucleus raphe magnus (NRM) on responses obtained from these units were investigated. Units responding to noxious mechanical stimulation of the integument with SPL input were found in the posterior shell region of the VPL. Responses elicited from these units by electrical stimulation of the SPL were inhibited following conditioning stimulation in or near the NRM. Inhibition could still be demonstrated after bilateral section of the dorsolateral funiculi at the level of C₃−C₄. Responses of these units to electrical stimulation of the ventrolateral funiculus (VLF) of the cervical cord were also inhibited following conditioning stimulation in or near the NRM. These results suggest that inhibition of these units produced by conditioning NRM stimulation may be partially mediated by an ascending pathway, in addition to the well-known descending spinal pathways. Glutamate stimulation of the NRM inhibited responses of nociceptive VPL units to SPL stimulation, but responses of the same units to VLF stimulation were little affected by the glutamate stimulation of the NRM. Inhibition of responses of nociceptive VPL units to SPL stimulation may be due to anti-dromic excitation of brainstem neurons having efferent connection with the NRM.     

Characterization of coeruleospinal inhibition of the nociceptive tail-flick reflex in the rat: Mediation by spinal α2-adrenoceptors

Several lines of evidence implicate bulbospinal noradrenergic pathways in antinociception and descending inhibition. In the present study, descending inhibition of the nociceptive tail-flick (TF) reflex by electrical stimulation in the dorsolateral pons (DLP) and the spinal neurotransmitter(s) mediating that inhibition were characterized in lightly pentobarbital-anesthetized rats. It was determined that 10 s of stimulation in the DLP prior to the application of heat to the tail resulted in optimum (lowest) thresholds for inhibition of the TF reflex. Conditioning-test studies indicated that the duration of the inhibitory effects produced by stimulation outlasted the 10-s period of stimulation by approximately 5 s. Systematic mapping studies revealed that inhibition of the TF reflex could be produced by stimulation throughout a large portion of the DLP; however, stimulation sites requiring the lowest intensities of stimulation (less than or equal to 25 microA) were in the locus coeruleus/subcoeruleus. Changes in blood pressure were not produced at this intensity and duration of stimulation. S-glutamate microinjections and stimulation strength-duration determinations suggest that inhibition of the TF reflex produced by stimulation in the locus coeruleus/subcoeruleus results from activation of cell bodies. The intrathecal administration of pharmacologic antagonists (phentolamine, yohimbine, prazosin, naloxone, methysergide, atropine and bicuculline) revealed that only the alpha-adrenergic antagonists phentolamine and yohimbine resulted in significant increases in stimulation thresholds in the locus coeruleus/subcoeruleus for inhibition of the TF reflex (83.1 and 93.9%, respectively). These results indicate that inhibition of the spinal nociceptive TF reflex produced by electrical stimulation in the locus coeruleus/subcoeruleus is at least in part a noradrenergic, postsynaptic alpha 2-adrenoceptor-mediated effect.     

Effects of neonatal capsaicin treatment on descending modulation of spinal nociception from the rostral, medial medulla in adult rat

Stimulation-produced modulation from the rostral, medial medulla (RMM) on the spinal nociceptive tail-flick (TF) reflex and on lumbar spinal dorsal horn neuron reponses to noxious cutaneous stimulti was studied in adult rats treated as neonates with capsaicin or vehicle. In vehicle-treated rats (n = 7), both descending facilitatory and inhibitory influences on the TF reflex were produced from the RMM. At 11/23 sites in the RMM, electrical stimulation produced biphasic modulatory effects. Electrical stimulation facilitated the spinal nociceptive TF reflex at low intensities (5–25 μA) and inhibited the TF reflex at greater intensities (50–200 μA). The mean threshold intensity of stimulation to inhibit the TF reflex (cut-off time= 7.0s) was 66 μA (n = 11). At 11 of 23 sites, electrical stimulation only inhibited the TF reflex; the mean threshold intensity of stimulation to inhibit the TF reflex was 50 μA (n = 11). At one stimulation site, electrical stimulation only facilitated the TF reflex at the intensities tested (5–100 μA). In capsaicin-treated rats (n = 6), the proportion of sites from which electrical stimulation only inhibited the TF reflex was significantly less (3/27sites= 11%) than in vehicle-treated rats (11/23= 48%). The threshold intensity of stimulation to inhibit the TF reflex from these three sites was 50 μA. The number of sites RMM fronm which electrical stimulation only facilitated the TF reflex was significantly greater in capaicin-treated rats (15/27= 56%) than in vehicle-treated rats (1/23= 4%). Neither the number of sites in RMM from which electrical stimulation produced biphasic modulatory effects on the TF reflex (48% and 33%, respectively) nor the intensities of stimulation or magnitudes of facilitation or inhibition of the TF reflex significantly differed between vehicle- and capsaicin-treated rats. In electrophysiological experiments, all units studied responded to non-noxious and noxious intensities of mechanical stimulation applied to the glabrous skin of the plantar surface of the ipsilateral hind foot and also to noxious heating of the skin (50°C). The number of sites where electrical stimulation produced only facilitatory effects on responses of spinal dorsal horn neurons to noxious stimulation (thermal or mechanical) of the skin was significantly increased from 13% of the total sites in vehicle-treated rats to 40% in capsaicin-treated rats. The number of sites where stimulation only produced inhibitory effects on responses of spinal dorsal horn neurons to noxious stimulation was decreased from 47% of the total sites in vehicle-treated rats to 35% in capsaicin-treated rats. The intensity of stimulation for producing inhibitory effects and the magnitude of inhibitory effects produced were not significantly different between vehicle- and capsaicin-treated rats. Immunocytochemical evaluation of the lumbar spinal dorsal horn verified that there were substantial reductions in substance-P and calcitonin gene-related peptide-like immunoreactivity, confirming the effectiveness of neonatal capsaicin treatment. The present study, in support of anatomical studies documenting central effects of capsaicin, suggests that neonatal capsaicin treatment also has significant effects on bulbospinal systems important to the modulation of spinal nociceptive transmission. Specifically, capsaicin treatment of neonates leads to a reduction in inhibitory and an increase in facilitatory influences on spinal nociception descending from the caudal brainstem.     

Serotonin and/or an excitatory amino acid in the medial medulla mediates stimulation-produced antinociception from the lateral hypothalamus in the rat

Several lines of evidence have demonstrated a role for the lateral hypothalamus (LH) in an endogenous system of descending inhibition. Descending inhibition from the LH relies, at least in part, on a relay(s) in the midbrain and/or medulla. The medullary nucleus raphe magnus (NRM) serves as one such relay. The present study, in rats lightly anesthetized with pentobarbital, was undertaken to systematically examine the transmitter(s) in the medial medulla mediating descending inhibition of the nociceptive tail flick (TF) reflex produced by focal electrical stimulation in the LH. The microinjection of pharmacologic receptor antagonists (5 μg) into the NRM revealed that the glutamate receptor antagonists, γ- -glutamylglycine and 2-amino-5-phosphonovalerate produced the largest increases in stimulation thresholds in the LH for inhibition of the TF reflex (107.6% and 102.6%, respectively). Methysergide, a serotonin receptor antagonist, also produced a significant increase (81.5%) in the stimulation threshold in the LH for inhibition of the TF reflex. The opioid receptor antagonist, naloxone, however, was without effect, producing only a 4.0% increase in the LH stimulation threshold. These results suggest that serotonin and/or an excitatory amino acid are transmitters at the bulbar relay in the medial medulla mediating descending inhibition of the TF reflex produced by focal electrical stimulation in the LH.     

Inhibition of spinal nociceptive transmission from the midbrain, pons and medulla in the rat: activation of descending inhibition by morphine, glutamate and electrical stimulation

It is generally believed that morphine activates a descending system(s) of inhibition, an effect contributing significantly to the analgesia produced. There has arisen, however, considerable controversy on this point. To address whether morphine inhibits spinal nociceptive transmission when given into the brainstem, the effects of focal electrical stimulation and monosodium S-glutamate (Glu) given in the periaqueductal gray (PAG), the locus coeruleus/subcoeruleus (LC/SC) and/or the nucleus raphe magnus (NRM) on spinal unit responses to noxious heating (50 °C) of the skin were examined and compared with effects produced by morphine (Mor). Focal electrical stimulation in 46 sites in the midbrain, dorsolateral pons and ventromedial medulla reliably inhibited unit responses to noxious heating of the skin (mean 34% of control). Microinjections of Glu (50 nmol, 0.5 μl) were made into 17 sites in the midbrain, 10 sites in the LC/SC and 11 sites in the NRM, inhibiting unit responses to a mean 57% at 22 of the 38 sites of microinjection. Mor (10–20 μg, 0.5 μl) was microinjected into 15 sites in the midbrain, 13 sites in the LC/SC and 11 sites in the NRM, inhibiting unit responses to heat to 63% of control at 24 sites of microinjection. The effects of morphine were shown to be receptor specific by antagonism with naloxone administered either intravenously or into the brainstem at the same site of microinjection as morphine. In 31 sites in the midbrain, dorsolateral pons and ventromedial medulla, microinjections of both Mor and Glu into the same sites attenuated unit responses to heating of the skin to a mean 77% and 71% of control, respectively. The results support the hypothesis that Mor acts supraspinally to modulate spinal nociceptive transmission by activating an endogenous descending inhibitory system(s). Focal electrical stimulation, glutamate and morphine modulated spinal nociceptive transmission by activation of descending inhibitory systems whose cell bodies of origin are in the PAG, the LC/SC or the NRM.     

Calcitonin: brainstem microinjection but not systemic administration inhibits spinal nociceptive transmission in the cat

In anaesthetized cats, nociceptive responses of lumbar dorsal horn neurons were studied during administration of salmon calcitonin (sCT). Systemic sCT administration (4–95IU/kg i.v.) produced no change in neuronal responses produced by noxious skin heating or by impulses evoked electrically in afferent C-fibres. Responses to skin heating were reduced during electrical stimulation in the brainstem, but the efficacy of this descending inhibition was not altered by systemic sCT administration. In contrast, noxious heat responses were clearly reduced by microinjection of sCT into the mesencephalic periaqueductal grey or the medullary raphe regions. These results suggest that calcitonin or a related peptide could act at specific brainstem sites to inhibit the spinal transmission of nociceptive information.     

Differential inhibition produced by peripheral conditioning stimulation on noxious mechanical and thermal responses of different classes of spinal neurons in the cat

The effect of conditioning stimulation of a peripheral nerve on responses of spinal neurons (dorsal horn cells and motoneurons) was studied in 16 decerebrate-spinal cats. The activity of dorsal horn cells was recorded with a microelectrode at the lumbosacral spinal cord and the single-unit activity of motoneurons was recorded from a filament of ventral rootlet divided from either the L7 or S1 ventral root. The responses of spinal neurons were evoked by noxious and innocuous mechanical stimuli and by noxious thermal stimuli applied to the receptive fields. The peripheral conditioning stimulation was applied to the tibial nerve with repetitive electrical pulses (2 Hz) at an intensity either suprathreshold for A delta or C fibers for 5 min. Applying conditioning stimulation to a peripheral nerve produced a powerful inhibition of the responses elicited by noxious stimuli, suggesting this inhibition is an antinociceptive effect. The inhibition produced by peripheral conditioning stimulation was differentially greater on the responses to noxious than to innocuous stimuli. Based on the results obtained from conditioning stimulation with graded strengths, afferent inputs from both myelinated and unmyelinated fibers seem to contribute to the production of the antinociceptive effect. The magnitude of the antinociceptive effect is bigger for the responses to noxious thermal than to mechanical stimuli. Furthermore, the reflex activity recorded in motor axons seemed to be more sensitive than in dorsal horn cells to the antinociceptive effect.     

Methysergide and supraspinal inhibition of the spinal transmission of nociceptive information in the anaesthetized cat

In anaesthetized cats methysergide was administered while measuring tonically present supraspinal inhibition of excitation of lumbar dorsal horn neurones by cutaneous noxious stimuli or by impulses in unmyelinated primary afferents. In some experiments inhibition was produced by electrical stimulation near the medullary raphe.Administered electrophoretically either near the bodies of dorsal horn neurones, in the region of the substantia gelatinosa, or topically to the dorsal surface of the spinal cord, methysergide did not reduce supraspinal inhibition.Administered intravenously in doses of up to 2.0 mg/kg, methysergide depressed the spinal transmission of nociceptive information and reduced supraspinal inhibition. It was considered that this latter effect did not result from a blockade of the action of 5-HT released by descending fibres but rather from a depression of the firing of the cells of origin.     

Inhibition of spinal cord interneurons by narcotic microinjection and focal electrical stimulation in the periaqueductal gray matter

Single cell evoked activity was recorded from spinal cord interneurons in rats prepared with microinjection cannulae or stimulating electrodes in the periaqueductal central gray matter (PAG). Morphine microinjections (4–16 μg) inhibited the response evoked by a noxious stimulus in 55% of the wide dynamic range neurons tested. Microinjections of etorphine (0.25–0.5 μg) inhibited 82% of the nociceptive neurons tested. Neither drug inhibited neurons which responded only to innocuous mechanical stimulation. The inhibition of wide dynamic range neurons produced by narcotic microinjection was antagonized by naxolone (1 mg/kg, i.p.) in 7 of 11 cases. Control experiments indicated that the effects obtained with microinjections could not be attributed to the drugs' diffusion to the spinal cord. Focal electrical stimulation of the PAG inhibited the responses to noxious stimuli of 60% of wide dynamic range neurons but was without effect on the responses of neurons that were activated only by innocuous stimuli. These experiments directly demonstrate that narcotic analgesics restricted to an intracerebral site of action activate a neural system which preferentially inhibits the responses of spinal cord wide dynamic range neurons to noxious stimuli. The system has a specificity for nociceptive input since non-nociceptive neurons were unaffected. Directly comparable results were produced by electrical stimulation of the PAG, supporting the concept that stimulation and narcotics modulate the transmission of nociceptive information by similar mechanisms.     

Opioid, cholinergic and α-adrenergic influences on the modulation of nociception from the lateral reticular nucleus of the rat

The lateral reticular nucleus (LRN) has been identified as an area in the caudal medulla involved in the centrifugal modulation of spinal nociceptive transmission and withdrawal reflexes. The data presented in this report further support a role for the LRN in the modulation of nociceptive responses. It was confirmed in the present study that focal electrical stimulation in the LRN inhibits the nociceptive tail-flick (TF) reflex at low intensities of stimulation in lightly pentobarbital-anesthetized rats. Aversive effects, however, were typically produced at similar and higher intensities of stimulation in the LRN in the same rats in the awake state. It was also determined that an inhibitory modulation of nociceptive responses organized both spinally and supraspinally could be activated independently by muscarinic cholinergic or opioid mechanisms in the LRN. Microinjection of morphine into the LRN in conscious rats produced an antinociception in both TF and hot plate (HP) tests which could be attenuated significantly by naloxone, but not atropine, previously microinjected into the same site in the LRN. Carbachol microinjected into the LRN also produced an antinociception which was attenuated significantly by atropine but not naloxone previously microinjected into the same site in the LRN. In contrast, the microinjection of clonidine or norepinephrine into the LRN either did not affect or shortened significantly response latencies in the TF and HP tests. These results further establish that the LRN contributes to the modulation of nociception. Opioid and cholinergic influences in the LRN appear to independently activate inhibition of responding to nociceptive stimuli organized either spinally or supraspinally, although descending inhibition was most clearly activated. An action at alpha 2 adrenoceptors in the LRN, conversely, produces an hyperalgesia as reflected by shortened latencies to respond in TF and HP tests.