Nucleus Raphe Magnus Stimulation Research Articles

Hindbrain raphe stimulation boosts cyclic adenosine monophosphate and signaling proteins in the injured spinal cord

Early recovery from incomplete spinal cord contusion is improved by prolonged stimulation of the hindbrain's serotonergic nucleus raphe magnus (NRM). Here we examine whether increases in cyclic adenosine monophosphate (cAMP), an intracellular signaling molecule with several known restorative actions on damaged neural tissue, could play a role. Subsequent changes in cAMP-dependent phosphorylation of protein kinase A (PKA) and PKA-dependent phosphorylation of the transcription factor “cAMP response element-binding protein” (CREB) are also analyzed. Rats with moderate weight-drop injury at segment T8 received 2h of NRM stimulation beginning three days after injury, followed immediately by separate extraction of cervical, thoracic and lumbar spinal cord for immunochemical assay. Controls lacked injury, stimulation or both. Injury reduced cAMP levels to under half of normal in all three spinal regions. NRM stimulation completely restored these levels, while producing no significant change in non-injured rats. Pretreatment with the 5-HT7 receptor antagonist pimozide (1mg/kg, intraperitoneal) lowered cAMP in non-injured rats to injury amounts, which were unchanged by NRM stimulation. The phosphorylated fraction of PKA (pPKA) and CREB (pCREB) was reduced significantly in all three regions after SCI and restored by NRM stimulation, except for pCREB in lumbar segments. In conclusion, SCI produces spreading deficits in cAMP, pPKA and pCREB that are reversible by Gs protein-coupled 5-HT receptors responding to raphe-spinal activity, although these signaling molecules are not reactive to NRM stimulation in normal tissue. These findings can partly explain the benefits of NRM stimulation after SCI.

Cortico-NRM Influences on Trigeminal Neuronal Sensation

We tested the idea that migraine triggers cause cortical activation, which disinhibits craniovascular sensation through the nucleus raphe magnus (NRM) and thus produces the headache of migraine. Stimulation of the dura mater and facial skin activated neurons in the NRM and the trigeminal nucleus. Stimulation of the NRM caused suppression of responses of trigeminal neurons to electrical and mechanical stimulation of the dura mater, but not of the skin. This suppression was antagonized by the iontophoretic application of the 5-HT(1B/1D) receptor antagonist GR127935 to trigeminal neurons. Migraine trigger factors were simulated by cortical spreading depression (CSD) and light flash. Activity of neurons in the NRM was inhibited by these stimuli. Multiple waves of CSD antagonized the inhibitory effect of NRM stimulation on responses of trigeminal neurons to dural mechanical stimulation but not to skin mechanical stimulation. The cortico-NRM-trigeminal neuraxis might provide a target for a more universally effective migraine prophylactic treatment.

Spatial and temporal patterns of serotonin release in the rat’s lumbar spinal cord following electrical stimulation of the nucleus raphe magnus

The monoamine neurotransmitter serotonin is released from spinal terminals of nucleus raphe magnus (NRM) neurons and important in sensory and motor control, but its pattern of release has remained unclear. Serotonin was measured by the high-resolution method of fast cyclic voltammetry (2 Hz) with carbon-fiber microelectrodes in lumbar segments (L3–L6) of halothane-anesthetized rats during electrical stimulation of the NRM. Because sites of serotonin release are often histologically remote from membrane transporters and receptors, rapid emergence into aggregate extracellular space was expected. Increased monoamine oxidation currents were found in 94% of trials of 50-Hz, 20-s NRM stimulation across all laminae. The estimated peak serotonin concentration averaged 37.8 nM (maximum 287 nM), and was greater in dorsal and ventral laminae (I–III and VIII–IX) than in intermediate laminae (IV–VI). When measured near NRM-evoked changes, basal monoamine levels (relative to dorsal white matter) were highest in intermediate laminae, while changes in norepinephrine level produced by locus ceruleus (LC) stimulation were lowest in laminae II/III and VII. The NRM-evoked monoamine peak was linearly proportional to stimulus frequency (10–100 Hz). The peak often occurred before the stimulus ended (mean 15.6 s at 50 Hz, range 4–35 s) regardless of frequency, suggesting that release per impulse was constant during the rise but fell later. The latency from stimulus onset to electrochemical signal detection (mean 4.2 s, range 1–23 s) was inversely correlated with peak amplitude and directly correlated with time-to-peak. Quantitative modeling suggested that shorter latencies mostly reflected the time below detection threshold (5–10 nM), so that extrasynaptic serotonin was significantly elevated well within 1 s. Longer latencies (>5 s), which were confined to intermediate laminae, appeared mainly to be due to diffusion from distant sources. In conclusion, except possibly in intermediate laminae, serotonergic volume transmission is a significant mode of spinal control by the NRM.

Differential opioid inhibition of C- and A delta- fiber mediated thermonociception after stimulation of the nucleus raphe magnus.

Although the importance of the nucleus raphe magnus in descending inhibitory control of nociception is clear, it is not known whether these effects are equivalent for different types of nociception. Thus, we examined the differential inhibition of behavioral responses evoked by A delta or C fiber thermonociceptor activation by electrical stimulation of nucleus raphe magnus neurons as well as the involvement of different classes of opiate receptors in this inhibition. In general, it was necessary to apply twice as much current to the nucleus raphe magnus to produce criterion antinociception for A delta mediated versus C fiber mediated nociceptive responses. Intrathecal administration of the nonselective opioid receptor antagonist, naltrexone, or the delta(1) opioid receptor antagonist, naltrindole, attenuated both A delta and C fiber antinociception induced by nucleus raphe magnus stimulation with similar efficacy. In contrast, intrathecal administration of naloxonazine, a micro specific opioid receptor antagonist, or naltriben, a delta(2) specific opioid receptor antagonist, preferentially attenuated nucleus raphe magnus induced antinociception for C fiber responses when compared with A delta mediated responses. These findings suggest that nociception evoked by the activation of A delta or C fiber nociceptors is under pharmacologically distinguishable descending control from the nucleus raphe magnus. Opiates differentially inhibit pain produced by the activation of myelinated or unmyelinated pain sensing neurons, a distinction that is clinically important. This article demonstrates that the brain's own pain control system operates with similar selectivity, and that this selectivity is partly mediated by different opiate receptor subtypes.

Physiological characteristics of the projection pathway from the medial preoptic to the nucleus raphe magnus of the rat and its modulation by the periaqueductal gray

Anatomical studies have shown a strong projection from the medial preoptic nucleus of the hypothalamus (MPO) to both the periaqueductal gray (PAG) and nucleus raphe magnus (NRM). In this study, we examined the physiological characteristics of MPO to NRM connections and examined how blockade of neuronal transmission and of the glutamatergic system within the PAG modifies this pathway. In deeply anesthetized rats, recordings were made from NRM neurons that were identified by their response to peripheral mechanical stimulation and designated as ‘E’, ‘I’, or ‘N’ if they were excited, inhibited, or not activated by noxious stimulation. In addition, cells were identified as spinally projecting if they could be antidromically activated by stimulation of the dorsolateral funiculus at the thoracic level. The responses of 204 NRM neurons to electrical and 87 cells to both chemical and electrical stimulation of MPO were recorded. The response of NRM neurons to MPO stimulation was highly dependent on the sensory class of these cells. Chemical stimulation of MPO inhibited 50% (16/32) and excited 16% (5/32) of the I-cells. In contrast, 23% (9/39) of the E-cells were inhibited and 49% (19/39) were excited by chemical stimulation of MPO. Electrical stimulation at intensities below 80 μA at 100 Hz had similar effects on the two classes of cells; 62% (24/39) of the E-cells and 31% (10/32) of the I cells were excited, and 31% (12/39) of the E-cells and 59% (19/32) of the I-cells were inhibited. The excitatory response to chemical stimulation lasted for an average of 136.8±73.2 s and inhibitory response lasted for an average of 143.8±102.1 s. Electrical stimulation of MPO at 1 Hz excited 27%, inhibited 3%, and had no effect on 70% of NRM cells. The mean latency to peak excitation was 9.6±6.6 ms. Antidromic activation of MPO neurons by NRM stimulation showed an average latency of 6.3±3.4 ms. Blocking the glutamatergic transmission within the PAG (by injecting kynurenic acid (KYN) into the PAG) blocked the inhibitory response of 40% (6/15) of the I-cells and inhibitory response of 43% (3/7) of the E-cells. The excitatory response of 27% (3/11) of the I-cells and the excitatory response of 14% (1/7) of the E-cells were blocked by kynurenic injection into the PAG. It is concluded that: (1) in response to chemical stimulation of MPO, the number of I-cells that were inhibited was more than three times the number of I-cells that were excited; in contrast, the number of E-cells that were excited was more than twice the number of E-cells that were inhibited. (2) The interaction between MPO and NRM can be modulated by blockade of the neuronal transmission or blockade of the glutamatergic system in the PAG. (3) Simultaneous activity of many synapses is required for activation of the MPO–NRM pathway. (4) MPO to NRM interaction is mediated by fibers with a conduction velocity of less than 1 m/s.

Central origins of substance P-like immunoreactive fibers and terminals in the spinal trigeminal caudal subnucleus in the rat

After trigeminal rhizotomy, some substance P-like immunoreactive (SP-LI) fibers and terminals in the spinal trigeminal caudal subnucleus (Vc), specially in its superficial laminae (laminae I and II), still remained in the rat. Employing a combination of Fluoro-Gold retrograde tracing and immunofluorescence histochemical staining for SP, we found that the main central origins of these SP-LI fibers and terminals were midbrain periaqueductal gray (PAG), nucleus raphe magnus (NRM) and other raphe nuclei, and nucleus reticularis gigantocellularis pars α; all of them are important structures of the endogenous pain control system. The present results provided morphological evidence for PAG or NRM stimulation could inhibit neuronal activities in the Vc evoked by orofacial nociceptive stimulation and also suggested that SP might be an important neurotransmitter or neuromodulator for endogenous pain control system.

Mechanism of transmission and modulation of renal pain in cats; effect of nucleus raphe magnus stimulation on renal pain.

Initially, when periaqueductal gray (PAG) is electrically stimulated, analgesia is induced, and this phenomenon is called stimulation-produced analgesia. Nucleus raphe magnus (NRM) as well as PAG are known to be the potent analgesic centers. NRM could modulate the nociceptive response of spinal cord neurons through spinally projecting fibers. However, as well as the above analgesic effects have been confined to the somatic pain, it was variable according to species, and the analgesic effect by NRM stimulation on the visceral pain was not yet clarified. In this study the analgesic effect by NRM stimulation on the visceral pain was examined through recording the activities of the dorsal horn neurons with renal input and renal pain, as a type of visceral pain. The renal pain was induced by ureteral occlusion or renal arterial occlusion, which in turn activated the renal mechanoreceptor or chemoreceptor. These cells had concomitant somatic input. In order to compare the effects of NRM stimulation on the renal pain with somatic pain, the somatic stimulation such as squeezing was conducted on the peripheral receptive field. The main results are summarized as follows: 1) After an electrical stimulation of NRM, spontaneous activities of dorsal horn neurons with renal input were reduced to 73.3 +/- 9.7% of the control value. 2) After an electrical stimulation of NRM, activities of dorsal horn neurons with renal input evoked by a brush, a type of non-noxious stimuli, did not change significantly. But the activities by a squeeze, a type of noxious stimuli, the activities were reduced to 63.2 +/- 7.2% of the control value. 3) After an electrical stimulation of NRM, activities of dorsal horn neurons with renal input evoked by occlusion of ureter or renal artery were reduced to 46.7 +/- 8.8% and 49.0 +/- 8.0% of the control value respectively. 4) The inhibitory effect of NRM on the dorsal horn neurons with renal input did not show any difference between renal A delta fiber and C fiber group. 5) By the electrical stimulation of NRM, the activities evoked by ureteral occlusion showed more reduction in the high threshold cell group than in the wide dynamic range cell group. These results suggest that activation of NRM can alleviate the renal pain as well as the somatic pain by modulating the dorsal horn neurons activities.

Parabrachial area and nucleus raphe magnus-induced modulation of nociceptive and nonnociceptive trigeminal subnucleus caudalis neurons activated by cutaneous or deep inputs.

1. The aim of this study was to test whether parabrachial area (PBA) stimulation exerts inhibitory influences on the spontaneous activity and responses evoked by skin and deep afferent inputs in trigeminal subnucleus caudalis (Vc) neurons, and to compare these effects with those of nucleus raphe magnus (NRM) stimulation. A total of 92 nonnociceptive and nociceptive Vc neurons was recorded in urethan/alpha-chloralose-anesthetized rats. Each neuron was functionally classified as low-threshold mechanoceptive (LTM), wide dynamic range (WDR), nociceptive-specific (NS), nociceptive convergent with both skin and deep inputs (S+D), or deep nociceptive (D); the LTM neurons could be subdivided as rapidly adapting (RA) or slowly adapting (SA). Conditioning stimulation was applied to histologically verified sites in PBA and NRM. 2. The spontaneous or evoked activity of all classes of neurons could be inhibited by PBA as well as by NRM stimulation, but generally the incidence and magnitude of inhibition were lower for the LTM neurons. Occasionally, facilitation of neuronal activity was also produced by PBA and NRM stimulation. 3. The spontaneous activity of 11 LTM neurons (6 RA, 5 SA), 13 nociceptive neurons (6 WDR, 7 NS), and 5 D neurons was tested with stimulation of PBA or NRM or both. LTM spontaneous activity was more significantly inhibited by NRM stimulation than by PBA stimulation, whereas both NRM and PBA stimulation had similar and significant inhibitory effects on NS, WDR, and D neurons. 4. The evoked nonnociceptive responses of 28 LTM neurons (16 RA, 12 SA) and of 6 WDR neurons were also tested with stimulation of PBA or NRM or both. The magnitudes of inhibition of the responses produced by PBA conditioning stimulation were statistically significantly less than those induced by NRM conditioning stimulation. 5. The cutaneous and deep nociceptive responses of cutaneous nociceptive neurons (9 NS, 19 WDR) and seven D neurons, respectively, were also tested with PBA and NRM stimulation. There was a significant difference in potency between PBA- and NRM-induced inhibition, but no difference in the magnitude of inhibitory effects among NS, WDR, and D neurons. For both PBA and NRM conditioning stimulation, graded increases in intensities of stimulation produced linear increases in inhibitory effects on nociceptive responses; an increase in stimulation frequency from 5 to 400 Hz also produced increases in inhibition of the nociceptive responses. 6. In five S+D nociceptive convergent neurons, the responses elicited by deep inputs were more powerfully inhibited by PBA stimulation than those elicited by cutaneous inputs.(ABSTRACT TRUNCATED AT 400 WORDS)

5-HT1A receptors mediate the effect of the bulbospinal serotonin system on spinal dorsal horn nociceptive neurons.

The present study examined whether the effect of stimulation of the nucleus raphe magnus (NRM) is mediated by spinal cord dorsal horn serotonin1A (5-HT1A) receptors in the rat. This hypothesis predicts that nociceptive dorsal horn units inhibited by NRM stimulation or iontophoretic 5-HT application would also be inhibited by iontophoresis of the selective 5-HT1A agonists 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and buspirone. A total of 78 dorsal horn wide-dynamic-range neurons were recorded. Overall, 62% of the cells tested (48/78) were responsive to electrical stimulation of the NRM with the predominant response being inhibitory (38/48; 79%). Fifty-eight cells were tested for their response to both NRM stimulation and 8-OH-DPAT iontophoresis: 20/58 cells were inhibited by NRM stimulation and 50% of the cells inhibited by NRM stimulation were also inhibited by 8-OH-DPAT. Fifty-two cells were tested for their response to both NRM stimulation and buspirone iontophoresis: 14/52 cells were inhibited by NRM stimulation with 9/14 similarly inhibited by buspirone. To examine whether exogenously applied serotonin produced an effect through 5-HT1A receptors, the effect of both 5-HT and 8-OH-DPAT iontophoresis was tested on 57 dorsal horn neurons. The majority of cells (25/57) were inhibited by 5-HT application; 15/25 were similarly inhibited by 8-OH-DPAT. The response of 48 dorsal horn cells to 5-HT and buspirone iontophoresis was compared. Forty-four percent (21/48) of the cells were inhibited by 5-HT; 16/21 were also inhibited by buspirone.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of norepinephrine in the interaction between the lateral reticular nucleus and the nucleus raphe magnus: an electrophysiological and behavioral study

We have previously demonstrated that the nucleus raphe magnus (NRM) sends a predominantly inhibitory projection to the lateral reticular nucleus (LRN); however, the pharmacology of this pathway is not known. The purpose of this study was to examine the role of norepinephrine in the NRM-LRN system using both electrophysiological and behavioral techniques. Sixty-nine LRN cells were recorded extracellularly. Cells were tested for their response to noxious and innocuous peripheral stimulation applied to the dorsal body surface. The majority of cells were classified as wide dynamic range, with inhibition being the predominant response; receptive fields were located primarily on the tail and hind limbs. The effect of excitatory amino acid glutamate (GLU) administration into NRM (GLU-NRM) was tested on all 69 cells. GLU-NRM inhibited 55 of 69 LRN cells tested; 7 cells were excited and 7 cells did not respond. Thirty-nine LRN cells were tested for their response to norepinephrine (NE) iontophoretically applied in LRN (NE-LRN). Two distinct types of effects were noted. In 9 cells, both NE-LRN and GLU-NRM produced a strong inhibition, with the magnitude of effect between the 2 drugs significantly correlated. In a second group of cells (n = 12), GLU-NRM produced an inhibitory effect while NE-LRN had no effect on the cells' baseline firing rate. However, when the 2 drugs were applied simultaneously, NE-LRN blocked the inhibitory effects of NRM stimulation. The effect of the α 2-receptor antagonist yohimbine (YOH) on NRM-evoked responses was tested in 30 LRN cells. The majority of these cells were inhibited by GLU-NRM. Similar to the dichotomous effect noted by NE-LRN, YOH applied iontophoretically in LRN (YOH-LRN) had two predominant effects on NRM-produced inhibition. In 14 of 27 cells, YOH-LRN significantly potentiated the inhibitory effects of NRM stimulation by increasing the duration of the inhibitory epoch an average of 100 sec. In 7 of 27 cells, YOH directly applied in LRN partially antagonized NRM-evoked inhibition. In a second series of experiments, microinjection cannulas were placed within NRM and LRN in order to determine the effect of blocking α 2-receptor activity within LRN on NRM stimulation-produced analgesia in an intact animal. Administration of d,l-homocysteic acid in NRM resulted in a significant increase in baseline tail-flick latency of approximately 140%. Pretreatment with YOH (3 μg in 0.5 μl) in LRN resulted in a significant potentiation of this analgesic effect. These results indicate that NRM sends a predominantly inhibitory projection to the LRN that involves noradrenaline and that this pathway is functionally active in an awake, behaving animal.

Raphe-spinal and subcoeruleo-spinal modulation of temperature signal transmission in rats

1. 1.Extracellular recordings were made from dorsal horn cells responsive to temperature changes in the innocuous range (10–40°C) of scrotal skin temperature in the lumbosacral segment of urethane-anesthetized rats. The temperature responsive dorsal horn cells (65) were classified into two groups: warm-responsive cells (48) showing an increase in firing rate by skin warming, “warm-response”, and cold-responsive cells (17) showing an increase in firing rate by skin cooling, “cold-response”. The effects of electrical stimulation in the nucleus raphe magnus (NRM) and subcoeruleus region (SC) on their firing activity were studied. 2. 2.The effects of electrical stimulation in the NRM as well as in the SC were found to consist in partial or complete depression of warm- or cold-responses. The background activity mostly remained unaffected. 3. 3.Depression of the temperature response by NRM stimulation occured in most (95%) of warm-responsive cells, whereas SC stimulation depressed the majority (79%) of cold-responsive cells. 4. 4.In four warm-responsive cells, NRM-evoked depression of their temperature responses was partially antagonized by the serotonin antagonist methysergide (1 mg/kg, i.v.), suggesting the NRM-evoked depression was mediated serotonergic raphe-spinal pathways. 5. 5.The results suggest that the raphe- and subcoeruleo-spinal descending pathways modulate the spinal transmission of warm and cold signals contributing to central thermoregulation.

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 C 3−C 4. 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.

Modulation of cortical evoked potentials by stimulation of nucleus raphe magnus in rats.

1. In pentobarbital sodium-anesthetized rats, we evaluated changes in cortical evoked potentials (EPs) associated with electrical and chemical stimulation of nucleus raphe magnus (NRM). A condition-test (C-T) paradigm was used. Cortical EPs were produced by test stimuli delivered to a hindpaw or the thalamic ventral posterior lateral nucleus (VPL; electrical stimulation), or by photic stimulation of the eyes or electrical stimulation of contralateral homotopical cortex (transcallosal EPs). These test stimuli were then preceded by electrical or chemical conditioning stimulation (CS) delivered to NRM through a stereotaxically implanted electrode or injection cannula, respectively. Effects of CS on EPs produced by the test stimuli were characterized. 2. Electrical CS preceding a test stimulus delivered to the foot reduced the amplitude of EPs at thresholds as low as 10-25 microA. The magnitude of EP reduction was dependent on CS intensity, frequency, and the C-T interval. Optimal parameters were trains of 10 pulses (400 Hz) delivered at a C-T interval of 5-10 ms. Injection of glutamate and lidocaine into NRM demonstrated that these effects were due to activation of NRM neurons and not to current spread to medial lemniscus (ML). NRM CS also reduced cortical EPs produced by test stimulation in VPL but did not alter EPs from visual stimulation or from electrical stimulation of contralateral homotopical cortex. 3. These findings suggest that NRM CS attenuates EPs by inhibiting thalamic or thalamocortical afferent activity. Because NRM CS affected all components of the cortical EPs, the effect appears to involve alteration of general sensory activity and is not nociception specific.(ABSTRACT TRUNCATED AT 250 WORDS)

Modulation of neuron activity of the midbrain periaqueductal gray matter influenced by monoaminergic brainstem structures

A study was done on base activity and changes in base activity (BA) of neurons in periaqueductal gray matter (PAG) during stimulation of monoaminergic structures of the brainstem: the nucleus raphe magnus (NRM), the locus coeruleus (LC), and the substantia nigra (SN), in rats anesthetized with hexenal (200 mg/kg). Three types of PAG neurons that differed in BA structure were identified. NRM, LC and SN stimulation changed BA only in type III neurons. Stimulation of these structures evoked an increase in BA frequency in 11.0–14.5%, and inhibition in 31.0–47.5% of type III neurons. Simultaneous stimulation of two structures led to a marked drop in intensity of effects. A depressing effect on BA was always detected if stimulation of one of the structures suppressed BA. Stimulation of two structures, with one being the NRM, was most effective. The role of PAG in the organization of the brain-stem component of the antinociceptive mechanism is discussed.

Effects of electrical and chemical stimulation of nucleus raphe magnus on responses to renal nerve stimulation

Electrical stimulation of the nucleus raphe magnus (NRM) inhibits some somatic and visceral input at the spinal level. This study was designed to examine the effects of electrical and chemical stimulation of NRM on neuronal responses to afferent renal nerve (ARN) stimulation. In chloralose-anesthetized rats, electrical stimulation of ARN elicited predominantly excitatory responses in spinal gray neurons. In 10 neurons studied, electrical stimulation of the NRM elicited an inhibition of spontaneous activity of 8 neurons and inhibited evoked responses to ARN stimulation in 6 neurons. Microinjection of glutamate (5–10 nmol in 0.5–1 μl) into the NRM elicited an inhibition of spontaneous activity in 9 neurons, a facilitation in 6 neurons and no response in 8 neurons receiving ARN input. Responses evoked by ARN stimulation were inhibited in 12 neurons, facilitated in 4 neurons and not affected in 8 neurons. We conclude that renal input can be modulated at the spinal level by activation of the NRM and adjacent tissue. Furthermore, the inhibition of spinal gray neuronal responses elicited by stimulation of the NRM appears to be due, at least in part, to activation of fibers of passage since non-selective electrical stimulation is more efficacious than selective chemical stimulation of neuronal soma and dendrites.

Central antiaversive and antinociceptive effects of anterior pretectal nucleus stimulation: attenuation of autonomic and aversive effects of medial hypothalamic stimulation

Previous studies have shown that stimulation of the rat anterior pretectal nucleus (APtN) strongly depressed a spinal reflex to noxious heat without causing significant aversion or depression of other motor responses. It is not known if APtN stimulation can similarly reduce the aversiveness of electrical stimulation of the brain, nor is it known if APtN stimulation is itself rewarding or aversive. This study used a simple switch-off paradigm to examine the rewarding properties of APtN stimulation at different sites throughout the nucleus and also used the tail-flick test to determine if the stimulation produced antinociception. The effects of APtN stimulation on the behavioural and autonomic responses to electrical stimulation of the medial hypothalamus (MH) and the nucleus raphe magnus (NRM) were also examined. The results show that electrical stimulation of dorsal APtN was rewarding and also caused antinociception which lasted for 50 min. However, sites which gave the strongest reward were not necessarily those which gave the greatest antinociception, as these effects were not correlated. Electrical stimulation of ventral APtN induced only aversive effects. The aversive and autonomic effects of MH stimulation were significantly reduced by conditioning stimulation of dorsal APtN. However, the very similar escape and autonomic effects of NRM stimulation were unaffected by APtN stimulation. These results suggest that electrical stimulation of the dorsal parts of the APtN has positive rewarding properties as well as the well-known antinociceptive effects. The antiaversive effects of dorsal APtN stimulation may be due in part to the inhibition of central substrates of aversion as well as inhibition of sensory neurones.

Descending influences from nucleus raphe magnus on fusimotor neurone activity in rats

1. 1.Single fusimotor fibres were isolated in the ventral roots of lumbosacral segments of urethane-anaesthetized rats, and effects of electrical stimulation of the nucleus raphe magnus (NRM) on their spontaneous activity were investigated. The experiments were carried out in rats whose bilateral preoptic and anterior hypothalamic regions (PO/AH) were electrolytically destroyed to eliminate the influences of these regions to fusimotor activity. 2. 2.Of 44 fusimotor fibres studied, 38 (86%) were found to be affected by NRM stimulation. The effects of NRM stimulation were classified according to their response pattern: primary depression (D-type, n = 24), facilitation followed by depression (F-D-type, n = 5) and primary facilitation (F-type, n = 8). The most predominant effect of NRM stimulation upon fusimotor activity was characterized by a strong depression followed by a complete cessation of firing lasting either for a short period or for more than 30 min (D- and F-D-type). 3. 3.In three fusimotor fibres studied in the different preparations, it was observed that a NRM-evoked depression response was blocked by an intraperitoneal administration of a serotonin antagonist, p-chlorophenylalamine ( p-CPA) (10 mg/kg). 4. 4.The results indicate that the NRM exerts descending inhibitory or facilitatory influences on fusimotor neurones, and suggest that cold shivering is controlled by modulating fusimotor neurone activity via the serotonergic raphe-signal pathways.

A 5-HT 1-type receptor mediates the antinociceptive effect of nucleus raphe magnus stimulation in the rat

The serotonin-containing raphe-spinal pathway has been implicated as playing an important role in analgesia. Several studies, however, have reported the inefficacy of traditional serotonin receptor antagonists at reversing the antinociceptive action of electrical stimulation in the raphe. In the light of recent reports on the existence of several types of 5-HT receptors in rat spinal cord, the present study investigated the ability of two antagonists, selective for two different 5-HT receptors to reverse the effects of focal electrical stimulation of the raphe magnus nucleus in the rat. Electrical stimulation of this nucleus resulted in selective antinociceptive as well as non-selective inhibitory effects on dorsal horn neurones. Both these effects were blocked by the ionophoretic application of a 5-HT 1, but not a 5-HT 2 receptor antagonist. The study presents data supporting the role of a spinal 5-HT receptor in mediating stimulation-produced analgesia from the nucleus raphe magnus and further, furnishes evidence that the 5-HT 1 receptor is involved in antinociception at the spinal level.

Effect of nucleus raphe magnus stimulation on recurrent inhibition of the monosynaptic reflex in the cat

Experiments were performed on 8 cats anesthetized with urethan-chloralose. The effects of nucleus raphe magnus (NRM) conditioning stimulation on recurrent inhibition of posterior biceps semitendinosus nerve (PBSt) monosynaptic reflex elicited by electric stimulation of a part of L7 ventral root were investigated in the cat. It was found that (1) the PBSt monosynaptic reflex was facilitated by NRM conditioning stimulation at 30, 50 and 80 ms conditioning-test stimulus intervals, but that (2) the inhibited monosynaptic reflex by recurrent inhibition was further inhibited by NRM conditioning stimulation at the same conditioning-test stimulus intervals, and that (3) the activity of Renshaw cells as recorded by glass microelectrode was enhanced by NRM conditioning stimulation. These facts indicated that recurrent inhibition pathways are enhanced by NRM conditioning stimulation.

Changes in mechanoreceptive field properties of trigeminal somatosensory brainstem neurons induced by stimulation of nucleus raphe magnus in cats

Experiments were carried out on adult anesthetized cats in which the effects of nucleus raphe magnus (NRM) conditioning stimulation (20 ms) were tested on the responses evoked by orofacial stimuli in single brainstem neurons of trigeminal (V) subnucleus oralis. The NRM stimulation induced inhibition of the responses of 57 of 77 low-threshold mechanoreceptive (LTM) neurons and the one wide-dynamic range (WDR) neuron tested. The duration of the neuronal inhibition ranged from 300-600 ms and the mean threshold for inhibition ranged from 47.8 +/- 4.8 to 102.7 +/- 15 microA depending on the orofacial stimulation site (skin or tooth pulp) and form (mechanical or electrical) of cutaneous stimuli used to evoke neuronal responses. In 20 LTM neurons showing NRM-induced inhibition that were specifically examined for the effects of NRM stimulation on the mechanoreceptive field, one population (n = 11) showed shrinkage (mean 55 +/- 4.4% from control area) of the mechanoreceptive field while the remaining neurons (n = 9) showed no change in mechanoreceptive field size during NRM stimulation. The former group of neurons were also distinguished from the latter neurons by their significantly larger mechanoreceptive field and the activation of the majority of them by electrical stimuli applied outside their mechanoreceptive field. The responses of these neurons evoked by low-threshold inputs from the edge of the mechanoreceptive field were more sensitive to NRM conditioning stimulation than responses evoked from the mechanoreceptive field center, as judged by threshold, magnitude and duration of the NRM-induced inhibition. These findings underscore the sensitivity of LTM neurons to NRM influences. They also reveal a particular population of oralis neurons which have a differential sensitivity of low-threshold inputs evoked from the edge compared to the center of the mechanoreceptive field.