Trigeminal Nociceptive Neurons Research Articles

IL-1β signaling initiates inflammatory hypernociception

Acute inflammatory pain associated with tissue injury triggers a wide variety of cellular changes in neurons of the associated sensory ganglia. These responses are frequently accompanied by hyperalgesia (heightened painful response) and allodynia (painful response to non-painful stimuli). Multiple laboratories have proposed that these abnormal sensations following inflammation are due to enhanced excitability or sensitization of nociceptive primary afferent neurons or nociceptors (commonly known as peripheral sensitization). In turn, these pronounced physiologic effects are likely to contribute to spontaneously active or ectopic afferent discharge and activity dependent changes in the response pattern of second order neurons in the spinal cord dorsal horn or trigeminal brainstem nuclei. This causes the once normal inputs to begin to produce abnormal responses or sensations (spinal sensitization). Although it is clear that molecular and possibly anatomical changes in the spinal cord dorsal horn or the trigeminal subnucleus caudalis are responsible for some attributes of this abnormal nociceptive sensation, it remains a mystery as to what molecules and which anatomical site(s) are essential for maintaining the neuronal signals of ongoing tissue inflammation. Several inflammatory mediators (bradykinin, substance P, calcitonin gene-related peptide), cytokines (IL-1β, TNFα, IL-6), chemokines (MCP-1), growth factors (NGF, NT-3, GDNF), prostaglandin E2 (PGE2) and/or complement proteins are thought to play a central role in the inflammatory hypernociception at the site of the inflamed tissue. However, the actions of these inflammatory mediators are not limited to the peripheral site of inflammation and may effectively alter signaling between neurons and glial cells. It is relatively well-accepted that spinal glial activation and the subsequent production of interleukin-1β (IL-1) play a role in the creation and maintenance of pathological pain states (Watkins et al., 2007). However, as described in this issue of Brain, Behavior, and Immunity by the research team of Mamoru Takeda and colleagues (this issue), this pathological effect may not be limited to cells of the CNS. Indeed, the combination of satellite glial cells (SGCs) and IL-1β may also serve a distinct role in the heightened pain sensitivity. Anatomically, SGCs form a distinct sheath around nearly all individual sensory neurons in DRGs, contribute to the transport and metabolism of molecules such as glutamate, glucose and ascorbate (Hanani, 2005) and maintain ionic homeostasis including the extracellular potassium (K+) concentration (Orkand et al., 1966; Newman et al., 1984; Vit et al., 2008). Following inflammation or injury of peripheral tissue, SGC cells begin to exhibit increased expression of glial fibrillary acidic protein (GFAP) and IL-1β, responses similar to CNS glial cells (Takeda et al., 2007). Takeda and colleagues (Takeda et al., 2007) also demonstrated that administration of IL-1β to activated SGCs in vitro potentiates neuronal excitability in the trigeminal ganglion. In the experiments now published in this issue of Brain, Behavior, and Immunity, the authors significantly extend their in vitro observations to in vivo conditions by testing the hypothesis that nociceptive trigeminal neurons are selectively sensitized by an IL-1β paracrine mechanism and are directly involved in development of tactile hypernociception. Following induction of cutaneous inflammation, extracellular single unit recordings from trigeminal neuronal activity were collected using multibarrel electrodes. Activation of Il-1β receptors was tested using iontophoretically applied antagonists. The results indicate that inflammation increases the spontaneous activity of small diameter trigeminal neurons through the involvement of IL-1β receptors. Taken together, these unique in vivo results suggest that an interaction between nociceptors and SGCs is essential for sensing ongoing states of peripheral inflammation via IL-1β. These findings should now be used to better define potential targets for the development of novel analgesics.

A Role For Wind-Up in Trigeminal Sensory Processing: Intensity Coding of Nociceptive Stimuli in the Rat

Wind-up is a progressive, frequency-dependent increase in the excitability of trigeminal and spinal dorsal horn wide dynamic range (WDR) nociceptive neurons evoked by repetitive stimulation of primary afferent nociceptive C-fibres. The correlate of wind-up in humans is temporal summation, which is an increase in pain perception to repetitive constant nociceptive stimulation. Although wind-up is widely used as a tool for studying the processing of nociceptive information, including central sensitization, its actual role is still unknown. Here, we recorded from trigeminal WDR neurons using in vivo electrophysiological techniques in rats and assessed the wind-up phenomenon in response to stimuli of different intensities and frequencies. First, we found that the amplitude of C-evoked responses of WDR neurons to repetitive stimulation increased progressively to reach a peak, then consistently showed a stable or slightly decreasing plateau phase. Only the first phase of this time course fitted in with the wind-up description. Therefore, to assess wind-up, we measured a limited number of initial responses. Second, we showed that wind-up, i.e. the slope of the frequency-dependent increase in the response to C-fibre stimulation, was linearly correlated to the stimulus intensity. Intensities of brief C-fibre inputs were thus coded into frequencies of action potentials by second-order neurons through frequency-dependent potentiation of the evoked responses. Third, wind-up also occurred at stimulation intensities below the threshold for C-evoked responses in WDR neurons, suggesting that wind-up can amplify subthreshold C-fibre inputs to WDR neurons. This might account for the observation that sparse, subliminal, neuronal activity in nociceptors can become painful via central integration of neural responses. Altogether, the present results show that wind-up can provide trigeminal WDR neurons with the capability to encode the intensity of short-duration orofacial nociceptive stimuli and to detect subthreshold nociceptive input. Thus, not only may wind-up play a physiological role in trigeminal sensory processing, but its enhancement may also underlie the pathophysiology of chronic orofacial pain conditions.

Molecular Mechanisms of Sensitization of Pain-transducing P2X3 Receptors by the Migraine Mediators CGRP and NGF

Migraine headache originates from the stimulation of nerve terminals of trigeminal ganglion neurons that innervate meninges. Characteristic features of migraine pain are not only its delayed onset but also its persistent duration. Current theories propose that endogenous substances released during a migraine attack (the neuropeptide calcitonin gene-related peptide [CGRP] and the neurotrophin nerve growth factor [NGF]) sensitize trigeminal neurons to transmit nociceptive signals to the brainstem, though the mechanisms remain poorly understood. Recent studies indicate that acute, long-lasting sensitization of trigeminal nociceptive neurons occurs via distinct processes involving enhanced expression and function of adenosine triphosphate (ATP)-gated P2X3 receptors known to play a role in chronic pain. In particular, on cultured trigeminal neurons, CGRP (via protein kinase A-dependent signaling) induces a slowly developing upregulation of the ionic currents mediated by P2X3 receptors by enhancing receptor trafficking to the neuronal membrane and activating their gene transcription. Such upregulated receptors acquire the ability to respond repeatedly to extracellular ATP, thus enabling long-lasting signaling of painful stimuli. In contrast, NGF induces rapid, reversible upregulation of P2X3 receptor function via protein kinase C phosphorylation, an effect counteracted by anti-NGF antibodies. The diverse intracellular signaling pathways used by CGRP and NGF show that the sensitization of P2X3 receptor function persists if the action of only one of these migraine mediators is blocked. These findings imply that inhibiting a migraine attack might be most efficient by a combinatorial approach. The different time domains of P2X3 receptor modulation by NGF and CGRP suggest that the therapeutic efficacy of novel antimigraine drugs depends on the time of administration.

Dorsal horn NK1-expressing neurons control windup of downstream trigeminal nociceptive neurons

Windup is a progressive, frequency-dependent increase in the excitability of trigeminal and spinal dorsal horn wide dynamic range (WDR) nociceptive neurons to repetitive stimulation of primary afferent nociceptive C-fibers. Superficial dorsal horn neurokinin 1 receptor (NK1R)-expressing neurons were recently shown to regulate sensitization of WDR nociceptive neurons through activation of a defined spino-bulbo-spinal loop. However, the windup of WDR nociceptive neurons was not regulated through this loop. In the present study, we sought to identify the alternative circuit activated by dorsal horn NK1Rs that mediates WDR neuron windup. As a model we used the rat spinal trigeminal nucleus, in which the subnucleus oralis (Sp5O) contains a pool of WDR neurons that receive their nociceptive C-input indirectly via interneurons located in the medullary dorsal horn (MDH). First, we found that intravenous injection of NK1R antagonists (SR140333 and RP67580) produced a reversible inhibition of Sp5O WDR neuron windup. Second, we anatomically identified in the MDH lamina III a subpopulation of NK1R-expressing local interneurons that relay nociceptive information from the MDH to downstream Sp5O neurons. Third, using microinjections of NK1R antagonists during in vivo electrophysiological recordings from Sp5O WDR neurons, we showed that WDR neuron windup depends on activation of NK1Rs located in the MDH laminae I–III. We conclude that, in contrast to central sensitization that is controlled by a spino-bulbo-spinal loop, Sp5O WDR neuron windup is regulated through a local circuit activated by MDH lamina III NK1Rs.

Migraine Aura

Recurrent episodes of transient focal neurologic symptoms, known as aura, occur in association with migraine headache in about 11.9 million people in the United States. At present, the International Headache Society has recognized 3 "typical" auras: visual, sensory, and language. Increasing evidence from investigations in human subjects suggests that typical auras may be the clinical manifestation of a cortical spreading depression (CSD)-like phenomenon. Other studies have shown altered reactivity and processing within the cortices of migraineurs who experience an aura, which might render them more vulnerable to CSD-like events. Recent investigations also support the hypothesis that events intrinsic to the cerebral cortex are capable of activating trigeminal nociceptive neurons and of affecting the caliber of vascular structures innervated by them. A better understanding of the mechanisms underlying the aura may potentially lead to more effective therapies, which will aim at preventing migraine headaches before they start.

Delayed Upregulation of ATP P2X3 Receptors of Trigeminal Sensory Neurons by Calcitonin Gene-Related Peptide

Recent evidence indicates a key role for the neuropeptide calcitonin gene-related peptide (CGRP) in migraine pain, as demonstrated by the strong analgesic action of CGRP receptor antagonists, although the mechanisms of this effect remain unclear. Most trigeminal nociceptive neurons releasing CGRP also express ATP-activated purinergic P2X3 receptors to transduce pain. To understand whether the CGRP action involves P2X3 receptor modulation, the model of trigeminal nociceptive neurons in culture was used to examine the long-term action of this peptide. Although 79% of CGRP-binding neurons expressed P2X3 receptors, acute application of CGRP did not change P2X3 receptor function. Nevertheless, after 1 h of CGRP treatment, strong enhancement of the amplitude of P2X3 receptor currents was observed together with accelerated recovery from desensitization. Receptor upregulation persisted up to 10 h (despite CGRP washout), was accompanied by increased P2X3 gene transcription, and was fully prevented by the CGRP antagonist CGRP(8-37). Surface biotinylation showed CGRP augmented P2X3 receptor expression, consistent with confocal microscopy data indicating enhanced P2X3 immunoreactivity beneath the neuronal membrane. These results suggest that CGRP stimulated trafficking of P2X3 receptors to the cell-surface membrane. Using pharmacological tools, we demonstrated that this effect of CGRP was dependent on protein kinase A and PKC activation and was prevented by the trafficking inhibitor brefeldin A. Capsaicin-sensitive TRPV1 vanilloid receptors were not upregulated. The present data demonstrate a new form of selective, slow upregulation of nociceptive P2X3 receptors on trigeminal neurons by CGRP. This mechanism might contribute to pain sensitization and represents a model of neuronal plasticity in response to a migraine mediator.

Trigeminal nociceptors express TLR-4 and CD14: a mechanism for pain due to infection.

Although certain bacterial species appear to be risk factors for pain due to odontogenic infections, comparatively little is known about the potential mechanisms mediating this effect. In this study, we tested the hypothesis that trigeminal nociceptive neurons express the TLR4 or CD14 receptors, thus enabling sensory neurons to detect and respond to tissue levels of bacterial substances such as lipopolysaccharide (LPS). Immunohistochemical analyses of human and rat trigeminal neurons demonstrated that a capsaicin-sensitive subclass of nociceptors (defined by expression of TRPV1, a capsaicin receptor) expresses both TLR4 and CD14. Moreover, human dental pulp collected from patients with caries lesions demonstrated co-localization of TLR4 and CD14, with markers of peripheral sensory neurons. Collectively, these studies indicate that the capsaicin-sensitive subclass of trigeminal nociceptors expresses TLR4 and CD14. These results indicate that pain due to bacterial infections may result, in part, from direct activation of nociceptors by bacterial products such as LPS.

Chronic IL-1β Signaling Potentiates Voltage-Dependent Sodium Currents in Trigeminal Nociceptive Neurons

The proinflammatory cytokine interleukin-1beta (IL-1beta) mediates inflammation and hyperalgesia, although the underlying mechanisms remain elusive. To better understand such molecular and cellular mechanisms, we investigated how IL-1beta modulates the total voltage-dependent sodium currents (INa) and its tetrodotoxin-resistant (TTX-R) component in capsaicin-sensitive trigeminal nociceptive neurons, both after a brief (5-min) and after a chronic exposure (24-h) of 20 ng/ml IL-1beta. A brief exposure led to a 28% specific (receptor-mediated) reduction of INa in these neurons, which were found to contain type I IL-1 receptors (IL-1RI+) on both their soma and nerve endings. In marked contrast, after a 24-h exposure, the total sodium current was specifically increased by 67%, without significantly affecting the TTX-R component. This potentiation of INa was suppressed in the presence of selective inhibitors of protein kinase C and G-protein-coupled signaling pathways, thereby suggesting that INa can be modulated through multiple pathways. In summary, the potentiation of INa through chronic IL-1beta signaling in nociceptive sensory neurons may be a critical component of inflammatory-associated hyperalgesia.

Effect of electrical stimulation of vagal afferents on trigeminal nociceptive neurons in the intralaminar nucleus of the thalamus

Effect of electrical stimulation of vagal afferents on trigeminal nociceptive neurons in the intralaminar nucleus of the thalamus

視床髄板内核の三叉神経性侵害受容ニューロン活動に及ぼす迷走神経電気刺激の影響(大阪歯科大学大学院歯学研究科博士(歯学)学位論文内容要旨および論文審査結果要旨の公表)

視床髄板内核の三叉神経性侵害受容ニューロン活動に及ぼす迷走神経電気刺激の影響(大阪歯科大学大学院歯学研究科博士(歯学)学位論文内容要旨および論文審査結果要旨の公表)

Contrasting phenotypes of putative proprioceptive and nociceptive trigeminal neurons innervating jaw muscle in rat.

BackgroundDespite the clinical significance of muscle pain, and the extensive investigation of the properties of muscle afferent fibers, there has been little study of the ion channels on sensory neurons that innervate muscle. In this study, we have fluorescently tagged sensory neurons that innervate the masseter muscle, which is unique because cell bodies for its muscle spindles are in a brainstem nucleus (mesencephalic nucleus of the 5th cranial nerve, MeV) while all its other sensory afferents are in the trigeminal ganglion (TG). We examine the hypothesis that certain molecules proposed to be used selectively by nociceptors fail to express on muscle spindles afferents but appear on other afferents from the same muscle.ResultsMeV muscle afferents perfectly fit expectations of cells with a non-nociceptive sensory modality: Opiates failed to inhibit calcium channel currents (ICa) in 90% of MeV neurons, although ICa were inhibited by GABAB receptor activation. All MeV afferents had brief (1 msec) action potentials driven solely by tetrodotoxin (TTX)-sensitive Na channels and no MeV afferent expressed either of three ion channels (TRPV1, P2X3, and ASIC3) thought to be transducers for nociceptive stimuli, although they did express other ATP and acid-sensing channels. Trigeminal masseter afferents were much more diverse. Virtually all of them expressed at least one, and often several, of the three putative nociceptive transducer channels, but the mix varied from cell to cell. Calcium currents in 80% of the neurons were measurably inhibited by μ-opioids, but the extent of inhibition varied greatly. Almost all TG masseter afferents expressed some TTX-insensitive sodium currents, but the amount compared to TTX sensitive sodium current varied, as did the duration of action potentials.ConclusionMost masseter muscle afferents that are not muscle spindle afferents express molecules that are considered characteristic of nociceptors, but these putative muscle nociceptors are molecularly diverse. This heterogeneity may reflect the mixture of metabosensitive afferents which can also signal noxious stimuli and purely nociceptive afferents characteristic of muscle.

7 視床髄板内核の三叉神経性侵害受容ニューロン活動に及ぼす迷走神経電気刺激の影響(第495回大阪歯科学会例会)

7 視床髄板内核の三叉神経性侵害受容ニューロン活動に及ぼす迷走神経電気刺激の影響(第495回大阪歯科学会例会)

Animal Models of Chronic Pain: Effects of jaw extension in an animal model of temporomandibular disorder

Temporomandibular disorders (TMD) affect 10% of the North American population, and can cause debilitating pain and discomfort for its sufferers. Temporomandibular joint (TMJ) pain is unique in the study of orofacial pain as there are greater sensory disturbances in pain conditions involving deep tissues, such as the TMJ, rather than cutaneous tissues (Sessle and Hu, 1990). Additionally, it has been suggested that orofacial deep inputs are more effective in inducing neuroplastic changes in trigeminal brainstem nociceptive neurons when compared to orofacial cutaneous inputs (Zou et al., 1999). An animal model of chronic TMD has been established by injecting Complete Freund's Adjuvant (CFA) into the TMJ of Sprague Dawley rats. The model shows numerous similarities to the human condition, including a decrease in range of jaw motion, increased plasma extravsation (indicating a damaged endothelial barrier and a compromise to tissue integrity), decreased joint space and tissue pathology. The model also behaved in a predictable way when treated with a COX-2 inhibitor, meloxicam. To explore central changes in the model, we examined neurons in the trigeminal subnucleus caudalis for the internalization the NK-1 receptor. Vehicle and CFA treated animals received 3 minutes of an innocuous stimulus, jaw opening with a 100 N force. CFA treated (but not control) rats receiving this innocuous stimulus showed distinct internalization of the NK-1 receptor, indicating release of substance P. These results are interpreted to indicate that this animal model of TMD exhibits a nociceptive response to innocuous jaw opening. Support: CIHR and the University of Western Ontario.

The orofacial formalin test

The subcutaneous injection of formalin into the rat upper lip generates behavioral responses that last several minutes. The time course of the response is similar to what is observed following formalin injection into the paw, i.e. biphasic, with an early and short-lasting first phase followed, after a quiescent period by a second, prolonged (tonic) phase. The applied chemical stimulus (formalin) can be qualified as noxious since it produces tissue injury, activates Aδ and C nociceptors as well as trigeminal and spinal nociceptive neurons and is felt as painful in man. In addition, increasing the concentration of formalin causes a parallel aggravation of histological signs of tissue inflammation and injury. The measured behavioral response (face rubbing) is a relevant end-point: prolonged face rubbing is evoked by formalin but not saline injection and a positive relationship between the amplitude of the response and the formalin concentration is observed, at least up to 2.5%. At higher formalin concentrations, the use of other or additional end-points should be considered. Finally, the behavioral response in the orofacial formalin test is sensitive to various opioid and non-opioid analgesics. The orofacial formalin test can then be considered as a reliable way of producing and quantifying nociception in the trigeminal region of the rat.

Chronic pain-autonomic interactions.

The nociceptive and the autonomic nervous systems interact at the levels of the periphery, spinal cord, brainstem, and forebrain. Spinal and visceral afferents provide converging information to spinothalamic neurons in the dorsal horn and to neurons of the nucleus tractus solitarius and parabrachial nuclei. These structures project to areas involved in reflex, homeostatic and behavioral control of autonomic outflow and nociception. Clinical and experimental observations on migraine suggest that a general hyperexcitability could develop along nociceptive trigeminal neurons allowing the activation of descending pathways that facilitate pain processing or the suppression of pathways that slow down pain transmission. Slowly progressive dysfunction of central pain systems secondary to the repetition of attacks of the originating headache has been implicated in transforming the process from an episodic to a chronic condition. This concept of an irritable focus in the central nervous system is currently known as "central sensitization" and is a characteristic feature of visceral pain. Considering primary headache as the result of the interactions between the nociceptive and autonomic systems (visceral pain) can help us to understand the complex pathophysiology of these disorders and to explore new treatments.

Activation of neurons in trigeminal caudalis by noxious oral acidic or salt stimuli is not reduced by amiloride

We investigated the possible role of amiloride-sensitive ion channels of the ENaC/DEGenerin superfamily in the activation of trigeminal nociceptive neurons elicited by noxious chemical stimulation of the oral mucosa using two methodologies, single-unit recording and c-fos immunohistochemistry. In pentobarbital-anesthetized rats, single-unit recordings were made from neurons in superficial laminae of dorsomedial trigeminal subnucleus caudalis (Vc) that responded to noxious thermal and chemical stimuli applied to the dorsal tongue. Successive application of each of three chemicals (250 mM pentanoic acid, n=6 units; 250 mM citric acid, n=8; 5 M NaCl, n=6) evoked responses that were not affected following topical application of amiloride (1 mM). In separate experiments, pentobarbital-anesthetized rats received one of the following stimuli delivered to the dorsal tongue: 250 mM pentanoic acid ( n=6); 1 mM amiloride followed by 250 mM pentanoic ( N=6); 5 M NaCl ( n=5); or 1 mM amiloride followed by 5 M NaCl ( n=5). Two hours later they were perfused with 4% paraformaldehyde and the brain stems processed for c-fos immunoreactivity. Both pentanoic acid and 5 M NaCl evoked similar numbers and patterns of fos-like immunoreactivity (FLI) in dorsomedial Vc and other brain stem regions, with no significant difference in counts of FLI in animals pretreated with amiloride. These results suggest that amiloride-sensitive Na + channels are not essential in mediating the activation of intraoral trigeminal nociceptors.

Trigeminal antinociception induced by bicuculline in the periaqueductal gray (PAG) is not affected by PAG P/Q-type calcium channel blockade in rat

We have recently shown that injection of the P/Q-type (Ca v2.1/α 1A) calcium channel blocker, ω-agatoxin IVA, into the periaqueductal gray (PAG) facilitates meningeal dural stimulation-evoked trigeminal nociceptive processing. We injected the GABA A antagonist bicuculline into the PAG in addition to the agatoxin and observed bicuculline's effect on neurons responding to dural stimulation recorded in the trigeminal nucleus caudalis of rats in order to determine if P/Q channel-mediated changes acted through GABAergic mechanisms. The inhibition of trigeminal nociceptive neurons characteristic of bicuculline administered into the PAG was maintained in the presence of blocked PAG P/Q-type calcium channels. This suggests the PAG descending pain modulatory pathway is not affected by P/Q-type calcium channel blockade at the postsynaptic GABAergic inhibitory interneuron and the facilitation produced by agatoxin is mediated by another mechanism. These findings have implications for disorders involving the PAG or P/Q-type channels, such as migraine, in particular for the development of preventative treatments, suggesting GABAergic and voltage-gated calcium channels could be separately modulated.

Orphanin FQ produces gender-specific modulation of trigeminal nociception: behavioral and electrophysiological observations

The present study aimed to determine if orphanin FQ, an endogenous ligand for the opioid receptor like-1 receptor, produces gender-specific effects in the modulation of N-methyl- D-aspartate (NMDA)-evoked responses of trigeminal nociceptive neurons, and in the NMDA-induced nociceptive behavior. Single-unit extracellular recordings were made from nociceptive-specific and wide dynamic range neurons in the superficial and deeper dorsal horn of the medulla (trigeminal nucleus caudalis) in anesthetized (1.5 g/kg urethane) rats. In the proestrous female, orphanin FQ applied microiontophoretically produced facilitation of the NMDA-evoked responses in 50% (16/32) of nociceptive neurons, inhibition in 31% (10/32), and biphasic effects in 19% (6/32). In contrast, in the male, it inhibited the responses in 86% (18/21), and facilitated the responses in 14% (4/21). In ovariectomized animals, orphanin FQ inhibited the responses in 75% (9/12) of nociceptive neurons, facilitated the responses in 17% (2/12) and produced biphasic effects in 8% (1/12). In contrast, in estradiol-treated ovariectomized rats, it facilitated the responses in 46% (5/11), inhibited the responses in 36% (4/11) and produced biphasic effects in 18% (2/11). For behavioral studies, NMDA-induced scratching behavior was used to assess the effects of orphanin FQ. Twenty-eight male, ovariectomized and estradiol-treated ovariectomized rats were microinjected with NMDA (2 nmol in 10 μl) alone through a cannula implanted in the medullary region, while another 27 rats were microinjected with orphanin FQ (10 nmol in 10 μl) 10 min prior to giving NMDA. Orphanin FQ reduced the NMDA-induced nociceptive scratching behavior by 92% in the male, and by 96% in ovariectomized rats. In contrast, in estradiol-treated ovariectomized animals, orphanin FQ facilitated the NMDA-induced scratching behavior by 210%. We conclude from these studies that orphanin FQ is primarily pronociceptive in the female and primarily antinociceptive in the male. Furthermore, we suggest that estrogen is involved in generating the gender-specific effects of orphanin FQ.

Systemic morphine reduces the wind-up of trigeminal nociceptive neurons

We assessed the effects of intravenous morphine on the wind-up of nociceptive neurons of the spinal trigeminal nucleus oralis (Sp5O). Extracellular recordings of Sp5O nociceptive convergent neurons were performed in intact halothane-anesthetized rats. Wind-up of C-fiber-evoked responses was elicited by repetitive electrical stimulation (train of 16 shocks, 0.66 Hz) of their receptive field at C-fiber intensity (3 times the threshold). Wind-up was tested for its sensitivity to morphine (6 mg/kg,i.v.), and the specificity of the effects was verified with naloxone (0.4 mg/kg, i.v.). Nineteen convergent neurons displaying wind-up were recorded. Morphine reduced the wind-up of all but one. In five cases, notwithstanding a reduced wind-up, the neuronal response evoked by the first stimulus in the train (initial input) was unexpectedly increased. Naloxone always antagonized morphine inhibitory effects on the wind-up. When administered systemically, morphine reduced the wind-up of trigeminal nociceptive neurons. This inhibitory effect occurred independently of morphine's ability to affect the initial C-fiber-evoked input. Our findings support the idea that systemic morphine probably blocks wind-up by acting at opioid receptors located postsynaptically to nociceptive primary afferents.

Encoding of noxious stimulus intensity by putative pain modulating neurons in the rostral ventromedial medulla and by simultaneously recorded nociceptive neurons in the spinal dorsal horn of rats

Neurons in the nucleus raphe magnus and adjacent structures of the rostral ventromedial medulla (RVM) are involved in the control of nociceptive transmission. In the RVM the so-called on-cells are excited, and the so-called off-cells are inhibited, by noxious stimuli applied almost anywhere on the body surface, thus showing that they receive information from spinal and trigeminal nociceptive neurons. In deeply anesthetized rats, recordings were made from RVM neurons that resembled on- and off-cells (herein called putative on- and off-cells) in order to investigate (1) how they encode the intensity of thermal noxious stimuli (46–56°C) applied to a hindpaw, and (2) how their encoding properties relate to those of simultaneously recorded spinal neurons. In 49 of 98 cases, a graded increase in the stimulus temperature caused a monotonic decrease in the response latency of putative on-cells, putative off-cells and spinal neurons, while the response discharge rate monotonically increased for putative on-cells and spinal neurons and decreased for putative off-cells. In the majority of simultaneous recordings of RVM and spinal neurons, the latency and discharge rate of the putative on- or off-cell were highly correlated with the latency and discharge rate of the spinal neuron, and the stimulus/response slopes were similar. These results show that putative on- and off-cells can encode the stimulus intensity in terms of response latency and discharge rate, and suggest that such encoding closely reflects spinal neuronal encoding. This may be relevant for the transmission and modulation of pain information by RVM neurons.