Lumbar Spinal Dorsal Horn Neurons Research Articles

Role of spinal bombesin-responsive neurons in nonhistaminergic itch.

Intrathecal administration of the neurotoxin bombesin-saporin reduces or abolishes pruritogen-evoked scratching behavior. We investigated whether spinal neurons that respond to intradermal (ID) injection of pruritogens also respond to spinal superfusion of bombesin and vice versa. Single-unit recordings were made from superficial lumbar spinal dorsal horn neurons in anesthetized mice. We identified neurons with three search strategies: 1) ID injection of the nonhistaminergic itch mediator chloroquine, 2) spinal superfusion of bombesin, and 3) noxious pinch. All units were tested with an array of itch mediators (chloroquine, histamine, SLIGRL, BAM8-22), algogens [capsaicin, allyl isothiocyanate (AITC)], and physical stimuli (brush, pinch, noxious heat, cooling) applied to the hindlimb receptive field. The vast majority of chloroquine-responsive units also responded to bombesin. Of 26 chloroquine-sensitive units tested, most responded to SLIGRL, half responded to histamine and/or BAM8-22, and most responded to capsaicin and/or AITC as well as noxious thermal and mechanical stimuli. Of 29 bombesin-responsive units, a large majority also responded to other itch mediators as well as AITC, capsaicin, and noxious thermal and mechanical stimuli. Responses to successive applications of bombesin exhibited tachyphylaxis. In contrast, of 36 units responsive to noxious pinch, the majority (67%) did not respond to ID chloroquine or spinal bombesin. It is suggested that chloroquine- and bombesin-sensitive spinal neurons signal itch from the skin.

MrgC agonism at central terminals of primary sensory neurons inhibits neuropathic pain

Chronic neuropathic pain is often refractory to current pharmacotherapies. The rodent Mas-related G-protein-coupled receptor subtype C (MrgC) shares substantial homogeneity with its human homologue, MrgX1, and is located specifically in small-diameter dorsal root ganglion neurons. However, evidence regarding the role of MrgC in chronic pain conditions has been disparate and inconsistent. Accordingly, the therapeutic value of MrgX1 as a target for pain treatment in humans remains uncertain. Here, we found that intrathecal injection of BAM8-22 (a 15-amino acid peptide MrgC agonist) and JHU58 (a novel dipeptide MrgC agonist) inhibited both mechanical and heat hypersensitivity in rats after an L5 spinal nerve ligation (SNL). Intrathecal JHU58-induced pain inhibition was dose dependent in SNL rats. Importantly, drug efficacy was lost in Mrg-cluster gene knockout (Mrg KO) mice and was blocked by gene silencing with intrathecal MrgC siRNA and by a selective MrgC receptor antagonist in SNL rats, suggesting that the drug action is MrgC dependent. Further, in a mouse model of trigeminal neuropathic pain, microinjection of JHU58 into ipsilateral subnucleus caudalis inhibited mechanical hypersensitivity in wild-type but not Mrg KO mice. Finally, JHU58 attenuated the miniature excitatory postsynaptic currents frequency both in medullary dorsal horn neurons of mice after trigeminal nerve injury and in lumbar spinal dorsal horn neurons of mice after SNL. We provide multiple lines of evidence that MrgC agonism at spinal but not peripheral sites may constitute a novel pain inhibitory mechanism that involves inhibition of peripheral excitatory inputs onto postsynaptic dorsal horn neurons in different rodent models of neuropathic pain.

Decreased Expression and Role of GRK6 in Spinal Cord of Rats After Chronic Constriction Injury

Nerve injury and inflammation can both induce neuropathic pain via the production of pro-inflammatory cytokines. In the process, G protein-coupled receptors (GPCRs) were involved in pain signal transduction. GPCR kinase (GRK) 6 is a member of the GRK family that regulates agonist-induced desensitization and signaling of GPCRs. However, its expression and function in neuropathic pain have not been reported. In this study, we performed a chronic constriction injury (CCI) model in adult male rats and investigated the dynamic change of GRK6 expression in spinal cord. GRK6 was predominantly expressed in the superficial layers of the lumbar spinal cord dorsal horn neurons and its expression was decreased bilaterally following induction of CCI. The changes of GRK6 were mainly in IB4 and P substrate positive areas in spinal cord dorsal horn. And over-expression of GRK6 in spinal cord by lentivirus intrathecal injection attenuated the pain response induced by CCI. In addition, the level of TNF-α underwent the negative pattern of GRK6 in spinal cord. And neutralized TNF-α by antibody intrathecal injection up-regulated GRK6 expression and attenuated the mechanical allodynia and heat hyperalgesia in CCI model. All the data indicated that down-regulation of neuronal GRK6 expression induced by cytokine may be a potential mechanism that contributes to increasing neuronal signaling in neuropathic pain.

Ionotropic glutamate receptors contribute to maintained neuronal hyperexcitability following spinal cord injury in rats

In this study, we examined whether topical treatment of glutamate receptor antagonists attenuate hyperexcitability of lumbar spinal dorsal horn neurons following low thoracic hemisection spinal cord injury in rats. Four weeks after spinal hemisection, neuronal activity in response to mechanical stimuli applied on the peripheral receptive field was significantly increased in three different phenotypes of lumbar spinal dorsal horn neurons: wide dynamic range (WDR), low threshold (LT) and high threshold (HT). Topical application of MK-801 (NMDA receptor antagonist, 50 µg) significantly attenuated the activity of WDR, but not LT and HT neurons; whereas, NBQX (AMPA receptor antagonist, 0.5 and 1 µg) significantly attenuated neuronal activity in all three phenotypes of neurons (* p < 0.05). However, MCPG (group I/II metabotropic glutamate receptor antagonist, 100 µg) had no effect. The present study, in the context of previous work, suggests that ionotropic glutamate receptor activation play critical roles in the maintenance of neuronal hyperexcitability and neuropathic “below-level” pain behavior following spinal hemisection injury.

Territorial and extra-territorial distribution of Fos protein in the lumbar spinal dorsal horn neurons in rats with chronic constriction nerve injuries

This study aimed to examine the relationship between temporal and spatial expression patterns of Fos protein in the spinal dorsal horn neurons and thermal hyperalgesia behaviors in rats with chronic constriction injury (CCI) to the sciatic nerve. Our results demonstrated that Fos protein expression in the spinal dorsal horn neurons at L5 segment ipsilateral and contralateral to CCI of the sciatic nerve was significantly greater than in sham rats from days 10 to 30 postoperatively (PO 10d to 30d), and was concentrated on the injury (ipsilateral) side. Unlike the short-lived expression after tissue inflammation, laminae I to VI (especially laminae III/IV) displayed a persistent greater number of Fos-like immunoreactive (Fos-LI) neurons for at least 30 days after CCI of the sciatic nerve. After the increase in laminae III/IV, Fos-LI neurons tended to gradually increase in laminae I/II and V/VI at L5 segment from PO 2d to 30d, which were correlated with the heat hyperalgesia (48 °C) behaviors measured by paw withdrawal latency in CCI rats but not in sham rats. Interestingly, a persistent increase of Fos-LI neurons in laminae I to VI at L5 segment of the ipsilateral and contralateral sides and at the L1 segment that was out of the normal central terminations of the sciatic nerve suggested the probable presence of territorial and extra-territorial central sensitization or inadequate central nervous system (CNS) adaptive mechanisms. These findings may partly explain why abnormal pain sensations are sometimes distributed in a pattern that does not coincide with the territories of nerves or with the posterior roots of the peripheral nerve after injury.

Spinal Neurons that Possess the Substance P Receptor Are Required for the Development of Central Sensitization

In previous studies, we have shown that loss of spinal neurons that possess the substance P receptor (SPR) attenuated pain and hyperalgesia produced by capsaicin, inflammation, and nerve injury. To determine the role of SPR-expressing neurons in modulating pain and hyperalgesia, responses of superficial and deep lumbar spinal dorsal horn neurons evoked by mechanical and heat stimuli and by capsaicin were made after ablation of SPR-expressing neurons using the selective cytotoxin conjugate substance P-saporin (SP-SAP). Morphological analysis and electrophysiological recordings were made after intrathecal infusion of vehicle, saporin alone, or SP-SAP. SP-SAP, but not vehicle or SAP alone, produced an approximately 62% decrease in SPR-expressing neurons in the dorsal horn. Loss of SPR-expressing neurons diminished the responses of remaining neurons to intraplantar injection of capsaicin. Peak responses to 10 microg of capsaicin were approximately 65% lower in animals pretreated with SP-SAP compared with controls. Additionally, sensitization to mechanical and heat stimuli that normally follows capsaicin was rarely observed. Importantly, responses to mechanical and heat stimuli in the absence of capsaicin were not altered after SP-SAP treatment. In addition, nociceptive neurons did not exhibit windup in the SP-SAP-treated group. These results demonstrate that SPR-expressing neurons located in the dorsal horn are a pivotal component of the spinal circuits involved in triggering central sensitization and hyperalgesia. It appears that this relatively small population of neurons can regulate the physiological properties of other nociceptive neurons and drive central sensitization.

Expression of 5-HT 1A receptor mRNA in rat lumbar spinal dorsal horn neurons after peripheral inflammation

The present study observed the expression of the 5-hydroxytryptamine (5-HT) 1A receptor mRNA in the lumbar spinal dorsal horn neurons following carrageenan inflammation using in situ hybridization (ISH). We also studied the co-localization of 5-HT 1A receptor mRNA and γ-amino butyric acid (GABA) or enkephalin (ENK) immunoreactivities using a combined fluorescent ISH and immunofluorescent histochemical double-staining technique. The finding of this study demonstrated that 5-HT 1A receptor mRNA was widely distributed in the spinal dorsal horn with the highest density in laminae III–VI. Following carrageenan-induced inflammation, the 5-HT 1A receptor mRNA expression in all layers of ipsilateral dorsal horn was significantly enhanced, and the peak occurred after 8 h. Furthermore, the number of 5-HT 1A receptor mRNA and GABA or ENK immunoreactive double-labeled cells was also markedly increased 8 h after carrageenan injection. These findings suggested that following peripheral inflammation, the synthesis of 5-HT 1A receptor was increased in the lumbar spinal dorsal horn neurons, especially in spinal GABA and ENK neurons.

Temporal and spatial distribution of Fos protein in the lumbar spinal dorsal horn neurons in the rat with chronic constriction injury to the sciatic nerve

The temporal and spatial expression pattern of Fos protein in spinal dorsal horn neurons was examined by immunohistochemistry in rats with chronic constriction injury (CCI) to the sciatic nerve. In normal animals, a few Fos-immunoreactive (-IR) neurons were detected in the dorsal horn of the lumbar spinal cord. Following induction of CCI, a very large number of Fos-IR neurons appeared in the spinal dorsal horn, but a significant number of Fos-IR neurons were also observed in the contralateral dorsal horn where primary afferents of the injured sciatic nerve rarely project. Sham-operated animals also had a significant number of Fos-IR neurons in the dorsal horn bilaterally. The number of Fos-IR neurons reached its maximal level 1 day following placement of the ligatures (PO 1d). The ratio of the number of Fos-IR neurons in the ipsilateral dorsal horn to the contralateral dorsal horn, however, had its peak level 3 days following CCI (3.1-fold increase compared to the contralateral dorsal horn). The number of Fos-IR neurons in the dorsal horn gradually decreased, but increased again around PO 15d. On PO 30d, the number of Fos-IR neurons decreased and became comparable to that in normal animals. The present results indicate that the induction of Fos-IR neurons in the dorsal horn caused by CCI is biphasic and reaches its maximal level on PO 3d, near the time of hyperalgesia onset.

Cutaneous responsiveness of lumbar spinal dorsal horn neurons is reduced by general anesthesia, an effect dependent in part on GABAA mechanisms.

Extracellular activity was recorded from single spinal dorsal horn neurons in both chronic cat and acute rat models. This was done to define the effects of anesthesia on the processing of sensory information elicited by nonnoxious tactile stimulation of peripheral receptive fields (RFs). In the chronic cat model, baseline data were obtained in physiologically intact, awake, drug-free animals before anesthetic administration (halothane 1.0-2.0%). This made it possible to compare and contrast activity of each cell in the drug-free and anesthetized state. Halothane effects were confirmed in the acute rat model (anesthetized, spinally transected, and in some cases decerebrate). In addition, the gamma-aminobutyic acid-A (GABAA)-receptor antagonist picrotoxin (2 mg/kg) was administered intravenously to verify that the observed halothane effect on spinal dorsal horn neurons was mediated by an interaction with GABAA-receptor systems. Halothane effects on three separate measures of response to nonnoxious tactile stimuli were observed in the chronic cat model. Halothane produced a significant, dose-dependent reduction in the low-threshold RF area of the neurons studied. Halothane also caused a significant reduction in neuronal response to RF brushing (dynamic stimulus) and to maintained contact with the RF (static stimulus). A dose dependency was not observed with these latter two effects. Neurons with a predominant rapidly adapting response seemed to be less susceptible to halothane suppression than slowly adapting cells. In the acute rat model an increase in halothane caused a reduction in neuronal response similar to that seen in the cat. The intravenous administration of 2 mg/kg of picrotoxin by itself caused no significant change in RF size or response to brushing. However, the same amount of picrotoxin did cause a 50% reversal of the halothane-induced reduction in RF size without causing a significant change in the halothane effect on response to RF brushing. In contrast to work recently reported in a chronic sheep model, halothane causes a significant reduction in spinal dorsal horn neuronal response to tactile stimulation of peripheral RFs. This effect is caused by, in part, but not exclusively, to GABAA-neurotransmitter systems. However, the relative influence of GABAA systems may vary with the nature of the stimulus.

Biphasic modulation of spinal nociceptive transmission from the medullary raphe nuclei in the rat.

The modulatory effects of electrical and chemical (glutamate) stimulation in the rostral ventromedial medulla (RVM) on spinal nociceptive transmission and a spinal nociceptive reflex were studied in rats. Electrical stimulation at a total 86 sites in the RVM in the medial raphe nuclei (n = 54) and adjacent gigantocellular areas (n = 32) produced biphasic (facilitatory and inhibitory, n = 43) or only inhibitory (n = 43) modulation of the tail-flick (TF) reflex. At these 43 biphasic sites in the RVM, facilitation of the TF reflex was produced at low intensities of stimulation (5-25 microA) and inhibition was produced at greater intensities of stimulation (50-200 microA). At 43 sites in the RVM, electrical stimulation only produced intensity-dependent inhibition of the TF reflex. Activation of cell bodies in the RVM by glutamate microinjection reproduced the biphasic modulatory effects of electrical stimulation. At biphasic sites previously characterized by electrical stimulation, glutamate at a low concentration (5 nmol) produced facilitation of the TF reflex; a greater concentration (50 nmol) only inhibited the TF reflex. In electrophysiological experiments, electrical stimulation at 62 sites in the RVM produced biphasic (n = 26), only inhibitory (n = 26), or only facilitatory (n = 10) modulation of responses of lumbar spinal dorsal horn neurons to noxious cutaneous thermal (50 degrees C) or mechanical (75.9 g) stimulation. Facilitatory effects were produced at lesser intensities of stimulation and inhibitory effects were produced at greater intensities of stimulation. The apparent latencies to stimulation-produced facilitation and inhibition, determined with the use of a cumulative sum method and bin-by-bin analysis of spinal neuron responses to noxious thermal stimulation of the skin, were 231 and 90 ms, respectively. The spinal pathways conveying descending facilitatory and inhibitory influences were found to be different. Descending facilitatory influences on the TF reflex were conveyed in ventral/ventrolateral funiculi, whereas inhibitory influences were conveyed in dorsolateral funiculi. The results indicate that descending inhibitory and facilitatory influences can be simultaneously engaged throughout the RVM, including nucleus raphe magnus, and that such influences are conveyed in different spinal funiculi.

Responses of rat spinal dorsal horn neurons to intracutaneous microinjection of histamine, capsaicin, and other irritants.

To investigate the spinal processing of cutaneous pruritic and algesic stimuli, single-unit recordings were made from wide-dynamic-range-type lumbar spinal dorsal horn neurons in pentobarbital-sodium-anesthetized rats. Neuronal responses were recorded to mechanical and noxious thermal stimuli, as well as to microinjection (1 microl) of histamine (0.01-10% = 9 x 10(-1)-9 x 10(-4) M), capsaicin (0.1% = 3.3 x 10(-3) M), or other algesic chemicals into skin within the receptive field via intracutaneously placed needles. Most (84%) of the 89 neurons responded to intracutaneous (i.c.) microinjection of histamine with a brief phasic discharge followed by an afterdischarge of variable (s to min) duration. Ten minutes after i.c. microinjection of histamine (but not NaCl), there was a significant increase in the mean area of the low-threshold (but not high-threshold) portion of unit mechanical receptive fields. However, responses to graded pressure stimuli were not significantly affected after histamine. Responses did not exhibit significant tachyphylaxis when histamine microinjections were repeated at 5- or 10-min intervals. Unit responses significantly increased in a dose-related manner to microinjection of histamine at concentrations ranging across 4 orders of magnitude. Within 30 s after i.c. microinjection of the H1 antagonist cetirizine, unit responses to i.c. histamine delivered at the same skin site were significantly attenuated. Unit responses to histamine, as well as to noxious thermal stimulation, were significantly reduced after systemic administration of morphine (3.5 mg/kg i.p.) in a naloxone-reversible manner. Application of a mechanical rub, scratch, or a noxious heat stimulus during the unit's ongoing response to i.c. histamine produced a brief and marked excitation, often followed by a period of reduced ongoing discharge. Unit responses to histamine were markedly suppressed by electrical stimulation in the midbrain periaqueductal gray. Most (79%) histamine-responsive units tested also responded to i.c. microinjection of capsaicin. After the initial microinjection of capsaicin, subsequent responses to histamine and capsaicin microinjections were significantly reduced. Units also responded to i.c. ethanol (capsaicin vehicle) in a dose-related manner, and showed tachyphylaxis to repeated i.c. ethanol at 80% but not at 8%. The mean response to 80% ethanol was significantly smaller than to 0.1% capsaicin. All units tested also responded to topical application of mustard oil (50%) and i.c. serotonin (30 microg). The results are discussed in terms of theories that attempt to reconcile psychophysical and clinical observations of pain and itch sensation.

Behavioral and electrophysiological assessment of hyperalgesia and changes in dorsal horn responses following partial sciatic nerve ligation in rats

Behavioral and electrophysiological methods were used to investigate the hyperalgesia and allodynia, and functional changes in lumbar spinal dorsal horn neurons, in a model of neuropathic pain (Selzer et al. 1990) involving ligation of one-third to one-half of one sciatic nerve in rats. One and 5 weeks following ligation, there was a significant reduction in hind limb withdrawal latency to noxious radiant heat on the operated side and, to a lesser degree, on the unoperated side. By 16 weeks, heat withdrawal latencies were reduced about equally (∼ 40%) on both sides. Withdrawal threshold to mechanical pressure was markedly reduced within 1 week on the operated side, and decreased in a time-dependent manner on the unoperated side. Heat withdrawal latency and von Frey withdrawal thresholds were not significantly affected in sham-operated rats. The same rats were tested in a paradigm measuring the isometric force of hind limb withdrawals elicited by graded noxious contact heat stimuli (38–52°C, 5 sec). Withdrawal force increased monotonically with stimulus temperature starting at a threshold of ∼ 40°C. Stimulus-response functions were not significantly different between a sham-operated group and groups tested 5 (acute) and 16 weeks (chronic) after partial sciatic nerve ligation. Following behavioral testing, the animals were deeply anesthetized with pentobarbital sodium to allow electrophysiological recording of responses of single lumbar dorsal horn wide-dynamic range-type neurons to mechanical and noxious thermal stimulation of the hind paw. Recordings were made from 6 sham-operated rats (26 neurons ipsilateral and 31 contralateral to the operated leg), from 7 rats receiving sciatic nerve ligation 5 weeks previously (29 ipsilateral and 29 contralateral to ligation), and from 7 rats receiving partial sciatic ligation 16 weeks previously (18 ipsilateral, 29 contralateral to ligation). In several ligated rats we were unable to find heat-responsive neurons with cutaneous receptive fields on the hind paw ipsilateral to the ligation. For the neurons that were sensitive to heat, responses increased monotonically from a threshold of 40–42°C. Neuronal stimulus-response functions for heat were not significantly different between ipsi- and contralateral (to operated) sides in the sham, 5-week or 16-week post-ligation groups, or between sham and 5- or 16-week post-ligation groups. Mechanical receptive field areas were not significantly different between ipsi- and contralateral sides in the sham and 5-week post-ligation groups, or between sham and 5-week post-ligation groups. However, receptive field areas were significantly larger in the 16-week post-ligation group (both ipsi- and contralateral to ligation) compared to sham and 5-week post-ligation groups. The results suggest that allodynia may be associated with a chronic enhancement of neuronal mechanosensitivity, but that the thermal hyperalgesia is not associated with enhanced neuronal responsiveness or force of withdrawal.

Inhibition of spinal nociceptive neurons by microinjections of somatostatin into the nucleus raphe magnus and the midbrain periaqueductal gray of the anesthetized cat

The effects of somatostatin (SOM) after intravenous application and intracerebral microinjection into the medullary nucleus raphe magnus (NRM) or into the periaqueductal gray (PAG) on the spinal nociceptive transmission was quantitatively studied in the anesthetized cat. Noxious heat-evoked responses of multireceptive lumbar spinal dorsal horn neurons were reversibly depressed to 56.6 ± 9.7% of the control after systemically applied SOM (7 μg/kg i.v.; 7 μg/kg per h infusion rate). At 11 of 14 brainstem microinjection sites in the NRM and PAG, SOM (2.5 μg/μl) attenuated the heat-evoked responses to 58.9 ± 6.2% ( n = 5) (NRM) and 64.4 ± 6.3% ( n = 6) (PAG) of the control. After microinjection, maximal inhibition was reached within 8–14 min (NRM) or 23–29 min (PAG), respectively. Inhibition was reversible within 60 min after the injection. Thus, SOM has an antinociceptive potency by activating descending inhibition of nociceptive dorsal horn neurons from the NRM and PAG.

The inhibitory effect of substance P antagonist, CP-96,345, on the late discharges of nociceptive neurons in the rat superficial spinal dorsal horn

The effects of local spinal applications of the substance P antagonist, CP-96,345, on the electrically-evoked discharges of nociceptive neurons were examined in the superficial spinal dorsal horn of adult rats anesthetized with sodium pentobarbital. Extracellular single-unit recordings were made from lumbar spinal dorsal horn neurons which were excited by noxious mechanical stimulation and had early and late discharges evoked by A- and C-fiber inputs, respectively, following transcutaneous electrical stimulation. CP-96,345 was applied directly onto the surface of the spinal cord close to the recording sites. A dose of 50 nmol of CP-96,345 drastically inhibited the late discharges of 80% of neurons, without affecting early discharges significantly. This study demonstrated the specific inhibitory effect of CP-96,345 on the late discharges of a restricted population of spinal dorsal horn neurons. These results illustrate the important role of substance P in nociceptive transmission mediated by primary afferent C-fibers in normal acute pain.

Characteristics of propriospinal modulation of nociceptive lumbar spinal dorsal horn neurons in the cat

The segmental and laminar origin of propriospinal antinociceptive systems in the cat spinal cord and the modes to activate them are characterized. The experiments were performed on pentobarbital-anesthetized cats with a high cervical spinalization. Recordings were made from single lumbar spinal dorsal horn neurons responding to noxious radiant skin heating and to innocuous mechanical skin stimuli. The segemental and laminar origin of heterosegmental, propriospinal neurons modulating background activity and nociceptivc responses were identified and the conditions to activate them were characterized. Conditioning noxious front paw stimulation and superfusion of the cervical enlargement with l-glutamate, but not with substance P, reduced noxious heat-evoked responses of about 50% of all lumbar neurons tested. Glutamate superfusions of the lower thoracic or upper sacral spinal cord enhanced background activity and reduced nociceptive responses of most lumbar spinal dorsal horn neurons. Superfusions with substance P or somatostatin were ineffective. Glutamate microinjections into the superficial layers of the thoracic, upper lumbar or sacral dorsal horn ipsi- or contralateral to the recording sites or into lamina VIII of the ipsilateral thoracic or upper lumbar cord reduced noxious heat-evoked responses with or without changes in the level of background activity. It is concluded that propriospinal neurons originating from circumscribed areas of the cervical, thoracic, lumbar or sacral spinal cord independently modulate background activity and noxious heat-evoked responses of multireceptive lumbar spinal dorsal horn neurons. The incidence and efficacy of propriospinal antinociceptive stimulation sites was found to be as high as for the classical region of endogenous antinociception, the midbrain periaqueductal gray.

Effects of electrical stimulation of vagal afferents on spinothalamic tract cells in the rat

The effects of electrical stimulation of cervical vagal afferents (VAS) on the background activity and on the responses of 25 spinothalamic tract (STT) neurons to noxious stimuli were studied in anesthetized rats. Background (spontaneous) activity of 9 (36%) STT neurons was inhibited by all intensities of VAS, 6 (24%) units were facilitated at lesser and inhibited at greater intensities of VAS, 5 (20%) units were only facilitated by all intensities of VAS, and 5 (20%) units were not affected by VAS. Responses of 8 (36%) STT neurons to noxious stimuli were only inhibited by VAS, 9 (41%) were facilitated at lesser and inhibited at greater intensities of VAS, and 5 units (23%) were only facilitated by VAS. There were no significant differences in VAS-produced modulatory effects between STT neurons and 16 unidentified lumbar spinal dorsal horn neurons studied under the same conditions. These results reveal that descending facilitatory and inhibitory pathways engaged by activation of vagal afferents modulate rostrally projecting nociceptive transmission neurons in the spinal cord, constituting an important regulatory network for nociception.

Characteristics of midbrain control of spinal nociceptive neurons and nonsomatosensory parameters in the pentobarbital-anesthetized rat

1. GABAergic mechanisms in the midbrain periaqueductal gray (PAG) have been proposed to control the activity of descending antinociceptive systems and defensive behavior. Here, the effect of neuronal disinhibition by gamma-aminobutyric acid (GABAA) receptor blockade at midbrain sites on spinal neuronal responses to noxious and innocuous skin stimulation was quantitatively characterized. It was compared with the effect of direct neuronal excitation by glutamate microinjections or electrical stimulation at identical sites. Changes in mean arterial blood pressure and other nonsomatosensory responses were also assessed. 2. Responses of 101 multireceptive lumbar spinal dorsal horn neurons to noxious radiant skin heating (50 degrees C, 10 s), innocuous skin brushing, and electrical stimulation of primary afferent A- and C-fibers were recorded in deeply pentobarbital-anesthetized rats. The mean blood pressure was continuously monitored in one carotid artery, and other nonsomatosensory parameters, such as frequency and depth of spontaneous respiration and contractions of abdominal and facial muscles, were classified according to their relative intensity into five groups. 3. A fine, multibarrel glass pipette was constructed for monopolar electrical stimulation and microinjection of the GABAA receptor antagonist bicuculline (40, 200, or 400 pmol), or glutamate (10-50 nmol), or Fast Green dye in 50 or 100 nl at identical sites in the midbrain. 4. Bicuculline microinjections into discrete regions of the PAG selectively abolished spinal neuronal responses to noxious skin stimulation but did not affect brush-evoked responses or responses to electrical A-fiber stimuli. This antinociception was often, albeit not necessarily, accompanied by tachypnoea and abdominal and facial muscle contractions and changes--mostly increases--in mean arterial blood pressure. Injections into other areas of the PAG and adjoining ventral tegmentum (VT) were less effective. The vast majority of injection sites in the lateral tegmentum (LT) were ineffective. 5. Glutamate microinjections at midbrain sites to detect areas of origin of descending antinociceptive neurons were characterized by a high incidence (greater than 50%) of false-negative results, as bicuculline was shown to be effective at numerous glutamate-insensitive sites. Glutamate microinjections into some sites of the PAG and adjoining VT reduced, but did not abolish, spinal neuronal responses to noxious skin heating. Injections into the LT were ineffective. 6. The efficacy of electrical stimulation at midbrain sites on spinal nociceptive responses had no predictive value for the effect of glutamate or bicuculline.(ABSTRACT TRUNCATED AT 400 WORDS)

Spinal pathways mediating tonic or stimulation-produced descending inhibition from the periaqueductal gray or nucleus raphe magnus are separate in the cat.

1. The spinal pathways for tonic and stimulation-produced descending inhibition of spinal nociceptive neurons were investigated in anesthetized paralyzed cats. Reversible circumscribed blocks were produced at various depths in the lateral funiculi (LF) at L1-L2 using the microinjection of the local anesthetic lidocaine. The total amount of tonic descending inhibition in the absence of LF blocks was evaluated by monitoring the spinal neuronal activity during reversible spinalization by cold block and compared with the activity of the same neuron during LF blocks. Stimulation-induced descending inhibition of neuronal responses to noxious skin heating was produced by bipolar focal electrical stimulation in the periaqueductal gray (PAG) or nucleus raphe magnus (NRM) and compared with the inhibition of the same neurons during LF blocks. The relative significance of ipsi- and contralateral pathways in the dorsal, medial, or ventral aspects of the lateral funiculi for these types of descending inhibition are quantitatively described. 2. All 35 lumbar spinal dorsal horn neurons studied responded to noxious and innocuous mechanical and noxious thermal stimuli applied within the receptive fields on the glabrous skin of the hindlimb. Responses to noxious skin stimuli (50 degrees C, 10 s at 3-min intervals) were constant over time and served as a parameter to evaluate tonic and stimulation-produced descending inhibition. All neurons also responded to electrical stimulation of hindlimb cutaneous nerves supramaximal for the activation of A-beta-, delta-, and C-fibers. Neurons were located in laminae I-VI of the dorsal horn at L5-L7 levels. LF blocks were produced by the microinjection of 1 microliter lidocaine at each of one to six sites in the ipsilateral and/or contralateral LF 500, 1,500, and/or 2,500 microns below cord surface. 3. LF blocks ipsilateral to the recording sites in the cord significantly reduced tonic inhibition, with blocks in the dorsal part of the LF [i.e., the dorsolateral funiculus (DLF)] being equally effective to complete LF blocks. Stimulation-produced inhibition from PAG or NRM was, however, not significantly affected by ipsilateral LF blocks. 4. Contralateral LF blocks significantly reduced stimulation-produced descending inhibition and failed to affect tonic descending inhibition. Ventral LF blocks attenuated inhibition from the PAG but not from NRM, whereas DLF blocks were more effective on inhibition from the NRM. 5. Bilateral LF blocks significantly reduced tonic as well as stimulation-produced descending inhibition.(ABSTRACT TRUNCATED AT 400 WORDS)

Raphe magnus-induced descending inhibition of spinal nociceptive neurons is mediated through contralateral spinal pathways in the cat

In anesthetized cats, extracellular recordings were made from lumbar spinal dorsal horn neurons, driven by noxious radiant skin heating. Heat-evoked responses were inhibited during electrical stimulation in the medullary nucleus raphe magnus (NRM). To identify the spinal pathways mediating this descending inhibition, reversible blocks in the spinal cord white matter were produced by microinjection of the local anesthetic lidocaine. Descending inhibition from the NRM was significantly reduced during blocks in the dorsal and medial, but not ventral parts of the contralateral lateral funiculus (LF). Blocks at any site in the ipsilateral LF failed to affect NRM-induced descending inhibition. These results indicate that NRM-induced inhibition of nociceptive dorsal horn neurons is conveyed primarily in fibers descending in the contralateral spinal white matter.