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Distribution of Fos-like immunoreactivity in the caudal medullary reticular formation following noxious facial stimulation in the rat
Abstract
To investigate the topographic organization of nociceptive neurons in the caudal medullary reticular formation, the distribution of cells that exhibit c-fos expression was examined following a unilateral noxious facial stimulus: subcutaneous injection of formalin into the vibrissal pad of awake rats. Labelling for Fos-like immunoreactivity was present in a somatotopic distribution in a region of the lateral reticular formation adjacent to trigeminal nucleus caudalis, which corresponds approximately to lamina V of the medullary dorsal horn. Labelling in adjacent regions of the reticular formation showed no somatotopy but was predominantly ipsilateral. Contralateral labelling was concentrated ventrolaterally around the lateral reticular nucleus and dorsally near the nucleus of the solitary tract.
... Another approach that has been used is c-Fos labeling to delineate the arrangement of nociceptive neurons in the trigeminal spinal nucleus and upper cervical cord as well as spinal dorsal horn. After noxious stimulation of a variety of orofacial sites, Fos protein like-immunoreactive cells (Fos-LI cells) occur in the trigeminal spinal nucleus interpolaris (Vi) and caudalis (Vc) transition zone (Vi/Vc zone) and the Vc/C2 transition zone Iwata et al., 1998a;Zhou et al., 1999;Takemura et al., 2000;Nomura et al., 2002;Watanabe et al., 2002;Kawabata et al., 2004;Bereiter et al., 2005;Sato et al., 2005;Sugiyo et al., 2005Sugiyo et al., , 2006Shimizu et al., 2006) in a somatotopic arrangement, and their numbers increase as noxious stimulus intensity increases Mineta et al., 1995). However, it has been reported that the c-Fos expression starts at about 30 minutes and peaks about 1 hour after noxious stimulation (Hunt et al., 1987;Tokunaga et al., 1995) in both superficial and deep laminae of the spinal dorsal horn (DH; Bullitt, 1991), so it is very likely that c-Fos expression is not directly correlated with an activation of Vc and upper cervical nociceptive neurons following acute orofacial noxious stimulation. ...
... It has also been reported that the rostral Vc receives afferent inputs from rostral parts of the face, such as the nose and mouth, whereas cutaneous inputs from the lateral face and head project to the caudal part of the Vc (see Hu, 1990;Hu et al., 2005). Many electrophysiological studies have reported that the second-order neurons are somatotopically arranged in Vc according to the trigeminal primary afferent organization (Sessle and Greenwood, 1976;Price et al., 1976;Dubner et al., 1980Dubner et al., , 1989Dubner and Bennett, 1983;Hayashi et al., 1984;Sessle et al., 1986;Iwata et al., 1998bIwata et al., , 1999, and anatomical studies with c-Fos labeling have documented such a somatotopic arrangement of neurons in the Vc and upper cervical cord Mineta et al., 1995;Imbe et al., 1999;Gojyo et al., 2002;Nomura et al., 2002;Watanabe et al., 2002). The organization of nociceptive neurons in rat Vc and upper cervical spinal cord has been reported in single-neuron recording studies (see, e.g., Hu, 1990;Hu et al., 2005). ...
... It is probable that these fiber differences and technical limitation in single-neuron recording may be involved in the distribu- Generally, we observed a pattern of organization of pERK-LI cell expression following capsaicin stimulation of the face that was similar to that reported in previous c-Fos studies Zhou et al., 1999;Takemura et al., 2000;Watanabe et al., 2002;Nomura et al., 2002;Bereiter et al., 2005;Sato et al., 2005;Sugiyo et al., 2005Sugiyo et al., , 2006. However, the distribution of pERK-LI cells was more restricted from the Vi/Vc zone through the Vc/C2 zone compared with previous reports of c-Fospositive neurons Mineta et al., 1995). The time course of pERK expression documented in the present study reveals that the expression is a very early event occurring within 2 minutes and peaking at 5 minutes after a noxious facial stimulus. ...
To define the somatotopic arrangement of neurons in the trigeminal spinal subnucleus caudalis and upper cervical cord activated by acute noxious stimulation of various orofacial sites, pERK expression was analyzed in these neurons. After capsaicin injection into the tongue, lower gum, upper and lower lips, or mental region, pERK-like immunoreactive (pERK-LI) cells were distributed mainly in the dorsal half of the trigeminal spinal nucleus interporalis (Vi) and caudalis (Vc) transition zone (Vi/Vc zone), middle Vc, and Vc and upper cervical cord transition zone (Vc/C2 zone). pERK-LI cells were distributed throughout the dorsal to ventral portion of the Vi/Vc zone, middle Vc, and Vc/C2 zone following capsaicin injection into the anterior hard palate, upper gum, buccal mucosa, or vibrissal pad and in the ventral portion of the Vi/Vc zone, middle Vc, and Vc/C2 zone following snout, ophthalmic, or ocular injection of capsaicin. The rostrocaudal distribution area of pERK-LI cells was more extensive from the Vi/Vc zone to the Vc/C2 zone after intraoral injection than that after facial injection, and the rostrocaudal distribution of pERK-LI cells from the Vi/Vc zone to the Vc/C2 zone had a somatotopic arrangement, with the snout being represented most rostrally and ophthalmic, ocular, or mental regions represented most caudally. These findings suggest that the pERK-LI cells expressed from the Vi/Vc zone to the Vc/C2 zone following injection of capsaicin in facial and intraoral structures may be differentially involved in pain perception in facial and intraoral sites.
... In addition, we observed also a general contralateral activation in brainstem. It has been observed also in other studies (Mineta et al., 1995), considering that the mechanisms producing contralateral hyperalgesia are unknown but may be explained by the existence of interneurons connecting neurons of both sides (Koganemaru et al., 2000). There are several reports of trigeminal primary afferent projections to the contralateral subnucleus caudalis (Arvidsson and Gobel, 1981;Jacquin et al., 1990) In conclusion, our results suggest a key role for the endogenous ATP in peripheral pain processing involving the P2 receptors localized in skin tissue. ...
ATP plays an important role as an endogenous pain mediator generating and/or modulating pain signaling from the periphery to the central nervous system. The aim of this study was to analyze the role of peripheral purinergic receptors in modulation of the nitroxidergic system at a trigeminal ganglia level by monitoring changes in nitric oxide synthase isoforms. We also evaluated Fos-positive neurons in brainstem (spinal trigeminal nucleus) and pain-related behavior. We found that local administration of the P2 purinergic receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) decreased face-rubbing activity, nitric oxide synthase isoform expression in trigeminal ganglia, and Fos expression in spinal trigeminal nucleus after subcutaneous injection of formalin. These results suggest a role for peripheral P2 purinergic receptors in orofacial pain transmission through modulation of the nitroxidergic system. .
... Some neurones in this region receive convergent signals from vestibular nuclei and gastrointestinal afferents, and could mediate motion sickness-related vomiting (28,36). Importantly, subcutaneous injection of formalin into the vibrissal pad of awake rats and electrical stimulation of trigeminovascular afferents in the superior sagittal sinus of monkeys activates nuclei in the caudal part of the solitary tract (37,38). Furthermore, axons project from the nucleus tractus solitarius towards the dorsal medullary raphe and the trigeminal nucleus caudalis in humans (39). ...
The aim of this study was to determine whether scalp tenderness and photophobia, two well-recognized symptoms of migraine, develop during the motion sickness induced by optokinetic stimulation. To investigate whether motion sickness has a general influence on pain perception, pain was also assessed in the fingertips. After optokinetic stimulation, nausea increased more and headache persisted longer in 21 migraine sufferers than in 15 non-headache controls. Scalp tenderness increased during optokinetic stimulation in nauseated subjects, and pain in the fingertips increased more and photophobia persisted longer in migraine sufferers than controls. These findings suggest that the disturbance responsible for nausea also sensitizes trigeminal nociceptive neurones or releases inhibitory controls on their discharge. A low nausea threshold and a propensity for sensitization to develop rapidly in nociceptive pathways may increase susceptibility to migraine.
... Neurophysiological examinations in humans have demonstrated that peripheral trigeminal nerve lesions can cause R2 abnormalities in terms of delayed or absent responses (Aramideh and Ongerboer de Visser 2002), and we therefore suggest that a painful deafferentation of trigeminal afferent fibers may be a possible explanation of the group differences seen in the present R2 responses. However, because bilateral changes in R2 responses were found and no significant differences in R2 responses between the painful and the non-painful side, it seems likely that central connections between the right and left trigeminal nociceptive pathways could play a role (Mineta et al. 1995). ...
Atypical odontalgia (AO) is an orofacial pain condition which has been suggested to involve neuropathic pain mechanisms. The aim of this study was to use a brain stem reflex to investigate craniofacial nociceptive mechanisms in AO. In 38 AO patients and 27 matched healthy controls, the R2 component of the blink reflex (BR) was elicited using a "nociceptive-specific" electrode and recorded with surface electromyography electrodes on both orbicularis oculi muscles. The BR was tested by stimulation of both sides of the face of the participants before, during, and after an intraoral pain provocation test with capsaicin. The data were analyzed with three- and four-way mixed-model analyses of variance. The root mean square value of the ipsilateral R2 (R2i) was significantly reduced in patients compared with controls (P=0.046). No differences in R2 between stimulation sides were detected in either group (P>0.757). In all participants, R2 responses and the intensity of the pain evoked by the electrical stimulus were decreased during and after application of capsaicin compared with baseline (P<0.001). In patients, R2i onset latencies were significantly prolonged compared with controls (P=0.031). The present data show disturbances in the central processing of craniofacial information and that endogenous pain inhibitory systems in AO patients and healthy controls were activated to a similar degree by an acute intraoral nociceptive input. Additional clinical research with AO patients will be needed to determine to what extent neuropathic pain mechanisms are involved in this pain condition.
Abstract Introduction Psychophysical Studies of Oral Irritation Neurobiology of Oral Irritation Acknowledgments
NMDA receptors are associated to learning and memory processes, development and neural plasticity, as well as acute and chronic pain conditions. They are involved in the outset and maintenance of central sensitization associated to damage or inflammation of peripheral tissues. Glutamate is the main amino acid that stimulates the CNS, can be involved in nociceptive transmission processes at the spine level and is the main responsible for the fast synaptic transmission. The action of glutamate in the pain routes is mostly mediated by ionotropic receptors (AMPA, NMDA and kainic). The activation of NMDA receptors plays a major role in the excitatory neurotransmission and the synaptic plasticity of the CNS. Glutamate and its agonists (NMDA, AMPA or kainic acid) are involved in the generation and maintenance of hyperalgesia conditions (exacerbated response to noxious stimulus) and alodynia (decrease of pain threshold). The clinical effectiveness of NMDA receptor antagonists (ketamine, memantine, amantadine, dextromethorphan, methadone) is analyzed. Their potential indications include: cancer-related neuropathic pain; postherpetic neuralgia, chronic traumatism; amputation; spine injury; centrally-originated pain secondary to cerebrovascular accident, phantom limb pain; restless legs syndrome, orofacial chronic pain; fibromyalgia and surgery, among others. The effectiveness of ketamine when orally or parenterally administered has been studied in central dysesthesic pain, neuropathic pain in the traumatic cauda equina syndrome, alodynia and hyperalgesia. Dextromethorphan is a non-opiate antitussive and a non-competitive blocker of NMDA receptors. One of its indications is the management of diabetic neuropathy. Methadone binds mainly to opiate receptors mu, but it is also a non-competitive antagonist of NMDA receptors. The use of methadone improves the management of pain since it reduces the development of tolerance to opiates. Memantine is a non-competitive antagonist of NMDA receptors. It reduces the intracortical activation and increases the intracortical inhibition. Experimentally, it has shown antinociceptive effects during the phase secondary to the inflammation induced by the formaline test. It reduces the thermal and mechanical hyperalgesia. NMDA receptor antagonists open new perspectives for the management of neuropathic pain. These drugs are frequently combined with other analgesics, generally acting as coadjuvants. The disparate results obtained with different antagonists used for the same indication confirm the complexity of their mechanism of action, which is not limited to antagonism of NMDA receptors, but it goes much more beyond that. © 2004 Sociedad Española del Dolor. Published by Arán Ediciones, S.L.
Since the initial observation that peripheral noxious stimulation could induce expression of c-Fos in the spinal cord (Hunt et al., 1987), a myriad of studies have reported the activation of immediate early genes (IEGs) in response to nociceptive stimulation. Members of this class of genes are rapidly and transiently induced after cell stimulation, without the requirement of new protein synthesis. Given that IEG expression heavily depends on neuronal depolarization, the probing of both mRNA and protein products encoded by this class of genes have been extensively used to map neural circuits associated with a variety of experimental paradigms, including acute and chronic pain (Zimmermann and Herdegen, 1994; Herdegen and Zimmermann, 1995; Herdegen and Leah, 1998; Bester and Hunt, 2002). In fact, a variety of noxious stimuli, differing in type, duration and site of application have been shown to induce the expression of IEGs in both the spinal cord and brain (Zimmermann and Herdegen, 1994; Herdegen and Zimmermann, 1995; Herdegen and Leah, 1998; Bester and Hunt, 2002). These changes in gene expression have been proposed to mediate a cascade of events that ultimately lead to long-term alterations in neuronal functioning (Kaczmarek and Chaudhuri, 1997; Herdegen and Leah, 1998; Pinaud, 2004, 2005). Studying the induction of IEGs not only allows for the mapping of nociceptive pathways but also provides information about how abnormal activity within the central nervous system contributes to pathological pain. Increased pain sensitivity may arise from long-term changes in the excitability of neurons so that they are more sensitive to subsequent stimuli, and these changes may be mediated in part through IEG expression after noxious insult (Woolf and Salter, 2000; Scholz andWoolf, 2002). © 2006 Springer Science+Business Media, LLC. All rights reserved.
Neurons in the caudalmost ventrolateral medulla (cmVLM) respond to noxious stimulation. We previously have shown most efferent projections from this locus project to areas implicated either in the processing or modulation of pain. Here we show the cmVLM of the rat receives projections from superficial laminae of the medullary dorsal horn (MDH) and has neurons activated with capsaicin injections into the temporalis muscle. Injections of either biotinylated dextran amine (BDA) into the MDH or fluorogold (FG)/fluorescent microbeads into the cmVLM showed projections from lamina I and II of the MDH to the cmVLM. Morphometric analysis showed the retrogradely-labeled neurons were small (area 88.7 µm(2)±3.4) and mostly fusiform in shape. Injections (20-50 µl) of 0.5% capsaicin into the temporalis muscle and subsequent immunohistochemistry for c-Fos showed nuclei labeled in the dorsomedial trigeminocervical complex (TCC), the cmVLM, the lateral medulla, and the internal lateral subnucleus of the parabrachial complex (PBil). Additional labeling with c-Fos was seen in the subnucleus interpolaris of the spinal trigeminal nucleus, the rostral ventrolateral medulla, the superior salivatory nucleus, the rostral ventromedial medulla, and the A1, A5, A7 and subcoeruleus catecholamine areas. Injections of FG into the PBil produced robust label in the lateral medulla and cmVLM while injections of BDA into the lateral medulla showed projections to the PBil. Immunohistochemical experiments to antibodies against substance P, the substance P receptor (NK1), calcitonin gene regulating peptide, leucine enkephalin, VRL1 (TPRV2) receptors and neuropeptide Y showed that these peptides/receptors densely stained the cmVLM. We suggest the MDH- cmVLM projection is important for pain from head and neck areas. We offer a potential new pathway for regulating deep pain via the neurons of the TCC, the cmVLM, the lateral medulla, and the PBil and propose these areas compose a trigeminoreticular pathway, possibly the trigeminal homologue of the spinoreticulothalamic pathway.
Nitric oxide (NO) donors, which cause delayed headaches in migraineurs, have been shown to activate central trigeminal neurons with meningeal afferent input in animal experiments. Previous reports indicate that this response may be due to up-regulation of NO-producing cells in the trigeminal brainstem. To investigate this phenomenon further, we determined nitric oxide synthase (NOS)-containing neurons in the rat spinal trigeminal nucleus (STN), the projection site of nociceptive trigeminal afferents, following infusion of the NO donor sodium nitroprusside (SNP). Barbiturate anaesthetized rats were infused intravenously with SNP (50 microg/kg) or vehicle for 20 min or 2 h, and after periods of 3-8 h fixed by perfusion. Cryostat sections of the medulla oblongata containing the caudal STN were histochemically processed for detection of nicotineamide adenine dinucleotide phosphate (NADPH)-diaphorase or immunohistochemically stained for NOS isoforms and examined by light and fluorescence microscopy. The number of neurons positive for these markers was determined. Various forms of neurons positive for NADPH-diaphorase or immunoreactive to neuronal NOS (nNOS) were found in superficial and deep laminae of the STN caudalis and around the central canal. Neurons were not immunopositive for endothelial (eNOS) or inducible (iNOS) NOS isoforms. The number of NADPH-diaphorase-positive neurons increased time dependently after SNP infusion by a factor of more than two. Likewise, the number of nNOS-immunopositive neurons was increased after SNP compared with vehicle infusion. Around the central canal the number of NADPH-diaphorase-positive neurons was slightly increased and the number of nNOS+ neurons not changed after SNP treatment. NO donors increase the number of neurons that produce NO in the STN, possibly by induction of nNOS expression. Increased NO production may facilitate neurotransmitter release and promote nociceptive transmission in the STN. This mechanism may explain the delayed increase in neuronal activity and headache after infusion of NO donors.
A selection of World Wide Web sites relevant to reviews published in this issue of Current Opinion in Neurobiology.
Neurons with c-Fos protein-like immunoreactivity (fos-neurons) were examined in the rostral parts of the brainstem sensory trigeminal nuclear complex following intense electrical stimulation of the trigeminal nerves and noxious mechanical stimulation of the trigeminal receptive fields. Stimulation of all the examined nerves and receptive fields induced some fos-neurons at the medial edge of the subnucleus interpolaris but not in the principal sensory trigeminal nucleus. Stimulation of the primary neurons innervating the intraoral structures but not facial skin induced fos-neurons in the ipsilateral subnucleus oralis. These oralis fos-neurons were located in the dorsomedial nucleus that contained calcitonin gene-related peptide-like immunoreactivity. The oralis fos-neurons are considered to be involved in the processing of intraoral nociceptive signals.
c-fos and zif268 expression were assessed by immunocytochemistry for c-Fos and Zif268 proteins in the sensory trigeminal nuclear complex following noxious mechanical stimulation of the mandibular incisor pulp of rats. Marked up-regulation of both immediate early genes was observed in the subnucleus oralis ipsilateral to the stimulation. Cavity preparation of the dentine without reaching the pulp did not cause significant up-regulation detectable by immunocytochemistry. These results provide evidence that noxious dental signals reach the ipsilateral subnucleus oralis and up-regulate the transcription of immediate early genes c-fos and zif268.
To investigate the role of trigeminal subnucleus caudalis in neural mechanisms of irritation, we recorded single-unit responses to application of a variety of irritant chemicals to the tongue or ocular mucosa in thiopental-anesthetized rats. Recordings were made from wide dynamic range (WDR) and nociceptive-specific units in superficial layers of the dorsomedial caudalis (0-3 mm caudal to obex) responsive to mechanical stimulation and noxious heating of the ipsilateral tongue ("tongue" units) and from WDR units in ventrolateral caudalis (0-2 caudal to obex) responsive to mechanical and noxious thermal stimulation of cornea-conjunctiva and frequently also surrounding skin ("cornea-conjunctival" units). The following chemicals were delivered topically (0.1 ml) onto the dorsal anterior tongue or instilled into the ipsilateral eye: capsaicin (0.001-1% = 3.3 x 10(-2) to 3.3 x 10(-5) M), ethanol (15-80%), histamine (0.01-10% = 9 x 10(-1) to 9 x 10(-4) M), mustard oil (allyl-isothiocyanate, 4-100% = 4 x 10(-1) to 10 M), NaCl (0.5-5 M), nicotine (0.01-10% = 6 x 10(-1) to 6 x 10(-4) M), acidified phosphate buffer (pH 1-6), piperine (0.01-1% = 3.5 x 10(-2) to 3.5 x 10(-4) M), serotonin (5-HT; 0.3-3% = 1.4 x 10(-1) to 1.4 x 10(-2) M), and carbonated water. The dose-response relationship and possible tachyphylaxis were tested for each chemical. Of 32 tongue units, 31 responded to one or more, and frequently all, chemicals tested. The population responded to 75.3% of the various chemicals tested (</=10 per unit). The incidence of responses was independent of the order of chemicals tested, except for capsaicin, which reduced subsequent responses. Responses to histamine, nicotine, 5-HT, and ethanol had a more rapid onset and shorter duration compared with capsaicin, acid, and mustard oil. Responses to all chemicals increased in a dose-related manner. Successive responses to repeated application decreased significantly for nicotine, 5-HT, capsaicin, and piperine. Spontaneous firing increased significantly 5-10 min after initial application of capsaicin. Of 31 corneal-conjunctival units, 29 responded to one or more chemicals, and the population responded to 65% of all chemicals tested. Responses increased in a dose-related manner for all chemicals, and successive responses decreased significantly for histamine, nicotine, ethanol, acid, and capsaicin. Responses of tongue units to histamine and nicotine were reduced significantly by ceterizine (H1 antagonist) and mecamylamine, respectively. Mecamylamine also significantly reduced responses of corneal-conjunctival units to nicotine. Different classes of irritant chemicals contacting the oral or ocular mucosa can activate individual sensory neurons in caudalis, presumably via independent peripheral transduction mechanisms. Multireceptive units with input from the tongue or cornea-conjunctiva exhibited a similar spectrum of excitability to different irritant chemicals. Such neurons would not be capable of discriminating among different chemically evoked irritant sensations but could contribute to a common chemical sense.
This article reviews findings up to the end of 1997 about the inducible transcription factors (ITFs) c-Jun, JunB, JunD, c-Fos, FosB, Fra-1, Fra-2, Krox-20 (Egr-2) and Krox-24 (NGFI-A, Egr-1, Zif268); and the constitutive transcription factors (CTFs) CREB, CREM, ATF-2 and SRF as they pertain to gene expression in the mammalian nervous system. In the first part we consider basic facts about the expression and activity of these transcription factors: the organization of the encoding genes and their promoters, the second messenger cascades converging on their regulatory promoter sites, the control of their transcription, the binding to dimeric partners and to specific DNA sequences, their trans-activation potential, and their posttranslational modifications. In the second part we describe the expression and possible roles of these transcription factors in neural tissue: in the quiescent brain, during pre- and postnatal development, following sensory stimulation, nerve transection (axotomy), neurodegeneration and apoptosis, hypoxia-ischemia, generalized and limbic seizures, long-term potentiation and learning, drug dependence and withdrawal, and following stimulation by neurotransmitters, hormones and neurotrophins. We also describe their expression and possible roles in glial cells. Finally, we discuss the relevance of their expression for nervous system functioning under normal and patho-physiological conditions.
Although recent studies implicated glutamate receptors and nitric oxide in nociception, much still needs to be known about their localisation in neurons involved in nociceptive transmission from the orofacial region. In this study, c-fos expression indicated by Fos immunohistochemistry in the caudal spinal trigeminal nucleus induced by subcutaneous injection of formalin into the lateral face of the rat was used as a marker for nociceptive neurons. The study sought to determine whether Fos-positive neurons express nitric oxide synthase, glutamate N-methyl-D-aspartate type receptor subunit 1, and glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid type receptor subunit 2/3; and whether they project to the thalamus. After formalin injection, many Fos-positive nuclei appeared in the superficial laminae of the ipsilateral trigeminal nucleus. Confocal laser scanning microscope revealed that almost all neurons with Fos immunofluorescent nuclei were colocalised with N-methyl-D-aspartate receptor 1, 94% with glutamate receptor 2/3 and 14% with nitric oxide synthase. Some of them were closely related to neurons labelled by nitric oxide synthase. Lastly, some of the Fos-positive neurons were labelled by tetramethylrhodamine-dextran injected into the trigeminothalamic tract or the thalamic region. The results suggested that activation of N-methyl-D-aspartate receptor 1 and glutamate receptor 2/3 upon glutamate release in response to noxious stimulation to the orofacial region might mediate c-fos expression in neurons involved in nociception. The expression of Fos in the neurons could also be mediated by nitric oxide produced from the same, as well as neighbouring neurons, when nociceptive stimulation persisted. Fos-positive neurons in the spinal trigeminal nucleus may project to the thalamus, relaying orofacial nociception to the higher sensory centre.
To characterize the role of neuronal nicotinic acetylcholine receptors (nAChRs) in oral irritation and pain, we employed the method of c-fos immunohistochemistry to map the locations and numbers of brainstem neurons that express the immediate-early gene, c-fos, after application of nicotine to the tongue, either alone or after pretreatment with cholinergic antagonists. Groups of anesthetized rats received the following chemicals delivered bilaterally to the dorsal tongue: (1) 0.9% NaCl followed by nicotine (1%, 61 mM), (2) the nAChR antagonist, mecamylamine 0.1% (= 4.9 mM) followed by nicotine, (3) the muscarinic antagonist atropine (0.1% 1.46 mM) followed by nicotine, (4) atropine (1%, 14.6 mM) followed by nicotine, (5) 0.9% NaCl as a control, and (6) unstimulated controls. Two hours later, animals were perfused with phosphate-buffered saline followed by 4% paraformaldehyde through the aorta. Post-fixed brainstems were cut in 50-micron frozen sections and immunohistochemically processed for fos-like immunoreactivity (FLI). Following application of nicotine, there were significant increases in FLI compared with saline-treated controls in dorsomedial and ventrolateral aspects of the trigeminal caudalis. Pretreatment with either mecamylamine or the high (1%) concentration of atropine significantly reduced nicotine-evoked FLI in these areas, while pretreatment with the low (0.1%) atropine concentration did not significantly affect FLI. These results are consistent with the idea that nicotine activates nAChRs residing on lingual nociceptive fibers, which, in turn, excite neurons in trigeminal caudalis.
In order to investigate the c-fos response within the trigeminal nucleus caudalis (Sp5C) after noxious meningeal stimulation, capsaicin (0.25, 0.5, 1 and 5 nmol) was administered intracisternally in urethane (1 g/kg) and alpha-chloralose (20 mg/kg) anaesthetized male mice. Capsaicin induced a robust and dose-dependent c-fos-like immunoreactivity (c-fos LI) within Sp5C. C-fos LI was observed within laminae I and II of the entire brain stem from the area postrema to C2 level, being maximum at the decussatio pyramidum level. The area postrema, solitary tract, medullary and lateral reticular nuclei were also labelled. The 5-hydroxytryptamine(1B/1D/1F) receptor agonist sumatriptan (0.01, 0.1, 1 and 10 mg/kg), administered intraperitoneally 15 min before capsaicin stimulation (1 nmol), decreased the c-fos response within Sp5C, but not within solitary tract. The novel specific 5-hydroxytryptamine1F agonist LY 344864 (0.1 and 1 mg/kg, i.p.) significantly decreased the c-fos LI within the Sp5C as well. These findings suggest that intracisternally administered capsaicin activates the trigeminovascular system and that the pain neurotransmission can be modulated by 5-hydroxytryptamine(1B/1D/1F) receptors in mice. Thus, the availability of this model in mice, taken together with the possibility of altering the expression of specific genes in this species, may help to investigate further the importance of distinct proteins in the neurotransmission of cephalic pain.
To use the human blink reflex (BR) to explore possible neuropathic pain mechanisms in patients with atypical odontalgia (AO).
In 13 AO patients, the BR was elicited using a concentric electrode and recorded bilaterally with surface electromyographic (EMG) electrodes on both orbicularis oculi muscles. Electrical stimuli were applied to the skin above branches of the V1, V2, and V3 nerves and to the V branch contralateral to the painful branch. Sensory and pain thresholds were determined. The BR examination of the painful V branch was repeated during a capsaicin pain-provocation test. The data were analyzed with nonparametric statistics.
The BR responses (R2 and R3) evoked by stimulation of V3 were significantly smaller than the BR responses evoked by stimulation of V1 and V2 (P < .004). There were no differences in BR (R2 or R3) between the painful and nonpainful sides (P > .569), and the BR (R2 and R3) was not significantly modulated by experimental pain (P > .080). The sensory thresholds were significantly lower on the painful side compared to the nonpainful side (P = .014). The pain thresholds were not different between sides (P > .910).
No major differences between the V nociceptive pathways on the right and left sides were found in a relatively small group of AO patients. Future studies that compare BRs in AO patients and healthy volunteers are needed to provide further knowledge on the pain mechanisms in AO.
1. Recordings were made in anesthetized monkeys from neurons in the medullary reticular formation (MRF) caudal to the obex. Responses of 19 MRF neurons to mechanical, thermal, and/or electrical stimulation were examined. MRF neurons exhibited convergence of nociceptive cutaneous inputs from widespread areas of the body and face. 2. MRF neurons exhibited low levels of background activity. Background activity increased after periods of intense cutaneous mechanical or thermal stimulation. Nearly all MRF neurons tested failed to respond to heterosensory stimuli (flashes, whistle sounds), and none responded to joint movements. 3. MRF neurons were excited by and encoded the intensity of noxious mechanical stimulation. Responses to stimuli on contralateral limbs were greater than those to stimuli on ipsilateral limbs. Responses were greater to stimuli on the forelimbs than to stimuli on the hindlimbs. 4. MRF neurons responded to noxious thermal stimulation (51 degrees C) of widespread areas of the body. Mean responses from stimulation at different locations were generally parallel to those for noxious mechanical stimulation. Responses increased with intensity of noxious thermal stimulation (45-50 degrees C). 5. MRF neurons responded with one or two peaks of activation to percutaneous electrical stimulation applied to the limbs, the face, or the tail. The differences in latency of responses to stimulating two locations along the tail suggested that activity was elicited by activation of peripheral fibers with a mean conduction velocity in the A delta range. Stimulation of the contralateral hindlimb elicited greater responses, with lower thresholds and shorter latencies, than did stimulation of the ipsilateral hindlimb. 6. Electrophysiological properties of monkey MRF neurons resembled those of neurons in the medullary subnucleus reticularis dorsalis (SRD) in the rat. Neurons in the caudal medullary reticular formation could play a role in processing nociceptive information. Convergence of nociceptive cutaneous input from widespread areas of the body suggests that MRF neurons may contribute to autonomic, affective, attentional, and/or sensory-motor processes related to pain.
1. In anesthetized rats, recordings were made within the medullary subnucleus reticularis dorsalis (SRD) from neurons that exhibited convergence of nociceptive inputs from the entire body. Neurons with total nociceptive convergence (TNC) responded to suprathreshold percutaneous electrical stimuli (2-ms duration) with an early and a late peak due to activation of A delta- and C-fibers, respectively, no matter which part of the body was stimulated. Neurons with partial nociceptive convergence (PNC) responded to the same stimuli with an A delta-peak regardless of which part of the body was stimulated and with a C-peak of activation from some, mainly contralateral, parts of the body. The characteristics of the responses of these neurons to the application of graded intensities of electrical, thermal, and mechanical stimuli were analyzed. 2. All TNC neurons and 85% of PNC neurons responded to A delta- and C-fiber activation following percutaneous electrical stimulation of the contralateral hindpaw. With regard to A delta-fiber-evoked responses, a linear relationship between the logarithm of the applied current and the magnitude of the responses was found within the 0.25- to 6.0-mA and 0.5- to 24-mA ranges for TNC and PNC neurons, respectively; however, these curves were essentially similar. With regard to C-fiber-evoked responses, such a linear relationship was found within the 1.5- to 6.0-mA range for both types of SRD neurons, although the TNC neurons presented larger C-fiber-evoked responses than did the PNC neurons. 3. TNC and PNC neurons linearly increased their discharges during the application of noxious thermal stimuli to the contralateral hindpaw within the range 44-52 degrees C; the mean responses evoked by noxious heat from TNC neurons were of higher magnitude than those from PNC neurons. The majority of SRD neurons presented long-lasting afterdischarges, especially with the highest temperature employed (52 degrees C). 4. TNC neurons monotonically increased their discharges during graded mechanical or thermal stimulation of the tail. When mechanical stimuli were applied, a linear relationship was found between the logarithm of the strength of the mechanical stimulus and the neuronal discharges, in the 5.3- to 7.4-N/cm2 range. With thermal stimulation, TNC neurons linearly increased their discharges in the 44-52 degrees C range. When increasing amounts of the tail were immersed in a 50 degrees C waterbath, TNC neurons increased their discharges within a restricted range of tail surface areas (0.9-5.7 cm2); further increases in the stimulated surface size were not followed by increases in firing rate.(ABSTRACT TRUNCATED AT 400 WORDS)
In anesthetized rats recordings were made from neurons in the medulla caudal to the obex. In the medullary dorsal horn, typical trigeminal noxious specific, nonnoxious specific, and convergent neurons were found. In nucleus cuneatus, typical dorsal column units were recorded. In subnucleus reticularis dorsalis (SRD), recordings were made from neurons which exhibited convergence of nociceptive inputs from the entire body. In subnucleus reticularis ventralis (SRV) we recorded both from neurons with spontaneous activity that were either unaffected or inhibited by noxious stimuli applied to various parts of the body and from respiratory neurons. The present paper deals particularly with the nature of the stimuli that activated reticular neurons exhibiting nociceptive convergence.
The projections of the trigeminal nerve in the bulbar reticular formation of the cat have been investigated by recording unit activity after physiological stimulation of the face, in 50 "encéphales isolés" preparations. No somatotopical arrangement was found. The units susceptible to trigeminal afferents are widespread in the reticular formation of the medulla and are responsive to large areas of stimulation, mostly bilaterally symmetrical.
The lateral bulbar reticular formation of anesthetized rats was explored with microelectrodes in order to compare the response and field characteristics of its somatosensory cells with properties of units located in the adjacent spinal trigeminal complex. Responses were evoked in the cells using natural and electrical stimulation of the skin. Neurons in the reticular nuclei tended to exihibit long latencies, low folowing capacities and large receptive fields. On the other hand, trigeminal complex cells generally revealed opposite characteristics, more typical of primary sensory structures, such as short latencies, high following capacities and punctate-to-small fields. The distinction between the regions was emphasized by additional qualities which were peculiar to the reticular neurons: Bilateral peripheral representation, into very coarse mechnical stimulation. In amny respects, the data indicate that the lateral reticular formation is intimately involved in the mediation of facial pain. They also support the view that it participated in this function by regulating the flow of nocioceptive activity from receptive mechanisms of the skin to the lemniscal pathway.
The distribution of fos-like-immunoreactivity (fos-LI) in the medullary and upper cervical dorsal horn was examined following noxious facial stimulation, in order to evaluate the use of fos as a marker for neuronal activation in trigeminal nociceptive pathways. Control animals that received urethane anesthesia and no facial stimulation showed substantial bilateral labeling in the trigeminal complex that was restricted to one rostrocaudal level, at the transition between the medullary dorsal horn (nucleus caudalis) and nucleus interpolaris. Noxious mechanical stimulation (pinch) of different facial sites produced labeling in the ipsilateral dorsal horn whose distribution varied predictably with the rostrocaudal and dorsoventral position of the facial stimulation site, such that rostral facial sites were represented rostrally in the dorsal horn and dorsal sites were represented ventrolaterally. The cornea was exceptional among the facial stimulation sites in that it had a specific representation at two distinct rostrocaudal levels, in C1 and the interpolaris-caudalis transition region; the position of the rostral peak was somatotopically inappropriate, based on the representation of other facial sites. The proportion of labelling in laminae III–IV relative to laminae I–II was higher with noxious mechanical stimulation than with noxious thermal (55°C) or chemical (subcutaneous injection of capsaicin) stimulation. The proportion of labelling in laminae III–IV produced by electrical stimulation of the infraobital nerve was no greater than that produced by pinch.
The results suggest that fos-LI mapping can be a useful method for the investigation of somatotopy but is subject to serious limitations when used for the investigation of laminar organzation. The results also suggest that the interpolaris-caudalis transition region may have properties that are distinct from those of the rest of the trigeminal complex, possibly related to an involvement in autonomic function.
Immunocytochemical localization of the protein product of the proto-oncogene C-fos allows anatomical identification of physiologically activated neurons. The present study examined the subnuclear distribution of cFos protein in the rat caudal medulla following peripheral administration of cholecystokinin octapeptide, which reduces feeding and gastric motility by a vagally mediated mechanism. To begin phenotypic characterization of neurons activated to express cFos following cholecystokinin treatment, double-labeling techniques were used to identify vagal motor neurons and neurons immunoreactive for tyrosine hydroxylase, neuropeptide Y, and neurotensin. Activated cells were most prevalent in the subnucleus medialis of the nucleus of the solitary tract, less prevalent in the subnucleus commissuralis, and virtually absent in the subnuclei centralis and gelatinosus. Many activated cells occupied the caudal area postrema; some of these were catecholaminergic. In contrast, activated cells were sparse within the medial rostral area postrema. Other activated cells occupied the dorso- and ventrolateral medulla and the midline raphe nuclei. Retrograde labeling of vagal motor neurons confirmed that very few were activated. Those that were activated occupied the caudal dorsal motor nucleus. In the dorsomedial medulla, 51% of catecholaminergic neurons and 39% of neurons positive for neuropeptide Y were activated, but no neurotensin-positive neurons were activated. In the ventrolateral medulla, 25% of catecholaminergic neurons and 27% of neuropeptide Y-positive neurons were activated. By characterizing the subnuclear distribution and chemical phenotypes of neurons activated by exogenous cholecystokinin, these data contribute to elucidation of the neural circuits mediating the behavioral, physiological, and neuroendocrine effects produced by this peptide. © 1993 Wiley-Liss, Inc.
The stimulus modality required to excite single neurons was studied in the different portions of the trigeminal sensory complex. Regional preponderance of different modalities could be related to the mediolateral arrangement at all levels, but there was no apparent differentiation of the rostrocaudal axis with respect to modality, receptive field size, or adaptive properties. The directional sensitivity of neurons innervating specialized structures such as vibrissae and teeth is described. The few neurons which were excited by noxious stimuli were primarily isolated outside the morphological limits of the sensory V complex. It is concluded that virtually all trigeminal neurons can be excited by delicate tactile stimuli in the present experimental conditions. While presumably all tactile neurons can be influenced by painful stimuli, present evidence favors the absence of neurons uniquely sensitive to noxious stimuli. The representation of pain is interpreted as involving tactile neurons excited by small tactile fibers which are known to convey pain sensations under a variety of stimulus conditions.
Fifty-three per cent of 106 spontaneously active single neurones, located in the caudal medullary reticular formation of the urethane or halothane anaesthetized rat, were found to respond to a noxious stimulation, such as immersion of the tail in water at 52–53°C for 30 sec. The evoked responses consisted of either an increase (82%), decrease (7%) or biphasic (11%) alteration in firing rate, and could be produced by other noxious stimuli, such as a pinch, but not by stroking, tapping or joint manipulation. Morphine (0.8–1.0 mg/kg) or fentanyl (5–10 μg/kg) administered intravenously blocked the acceleration in firing rate produced by noxious stimuli, and this effect was antagonized by naloxone administered intravenously, suggesting it to be an action on specific opiate receptors. Naloxone (0.5–10 mg/kg) blocked those decreases in firing rate produced by noxious stimuli, suggesting that an endogenous opioid may be released onto these neurones as a result of noxious stimuli applied to the periphery.
1. A total of 113 trigeminothalamic neurons and over 200 presumed interneurons of nucleus caudalis (0-5 mm below the obex) and subjacent reticular formation were studied in rhesus monkeys anesthetized with chloralose or nitrous oxide. Each cell was characterized in terms of its antidromic responses to stimulation of ventral posterior medial and/or posterior thalamic nuclei and to three types of stimuli applied to its receptive field: a) graded 5-s temperature shifts at a rate of 9 degrees C/s from 35 degrees C to final temperatures of 20-52 degrees C, generated by a contact thermode; b) graded intensities of electrical stimulation to determine the conduction velocities of converging primary afferent fiber populations; and c) mechanical stimulation ranging from light touch to pinch with serrated forceps. 2. This analysis yielded five classes of units distinguished by the range of responses to mechanical stimuli and by the convergence of different primary afferent fiber populations. These five classes were found among both trigeminothalamic neurons and neurons which could not be antidromically activated. Class 1 units exhibited rapidly adapting responses to hair movement or light touch and received only A-beta primary afferent input. Class 2 units responded to light touch and pressure with maintained discharges and received A-beta primary afferent input. Class 3 units responded maximally to pinch with serrated forceps but also were activated by light touch and pressure. They received A-beta, A-delta, and C fiber input. Class 4 units responded to firm pressure and maximally to pinch with serrated forceps. These units had A-delta and sometimes C fiber input. Class 5 units responded only to pinch with serrated forceps and had exclusive A-delta fiber input. Some cells in all five classes responded antidromically to stimulation of the thalamus. Antidromic action-potential latencies of classes 1,2, and 3 units were shorter than those of classes 4 and 5 units (P less than 0.001). Receptive-field sizes were usually small (1-2 cm2) for classes 1, 2, 4, and 5 units, and larger for class 3 units (one to three trigeminal divisions). The marginal layer of nucleus caudalis contained mostly classes 4 and 5 units, some class 3 units, but no classes 1 or 2 units. The superficial portion of the magnocellular layer contained mostly classes 1 and 2 units, while neurons at the base of this layer contained class 3 units and some classes 4 and 5 units. Cells in the sujacent reticular formation included all 5 classes but showed a tendency to have large receptive fields (greater than 1 trigeminal division). 3. Neurons responding to noxious thermal stimuli (44-52 degrees C) were classes 3 or 4 units. The response patterns of classes 3 and 4 units to noxious thermal stimuli were similar. No classes 1 or 2 units and only one class 5 unit responded to increases in skin temperature. Thermal thresholds ranged from 38 to 50 degrees C and most heat-responsive units responded monotonically to temperatures between 45 and 52 degrees C...
Single units in the rat's brain stem that responded to tactile stimulation of the cornea were found mainly in a region on the ventromedial border of the trigeminal nuclei. This region included adjacent parts of the reticular formation and extended from the principal sensory trigeminal nucleus to caudal portions of nucleus caudalis of the spinal trigeminal complex. One class of units responded to light punctate stimuli and small electric currents; a second class had higher thresholds for both kinds of stimulation. The former units generally had shorter response latencies and larger corneal receptive fields than the latter units, although distributions for the two kinds of units overlapped appreciably for both properties. High-threshold and low-threshold units were found at all rostrocaudal levels and were not distributed differentially in the trigeminal nuclei or in the reticular formation. Units in a third class were scattered in more medial parts of the reticular formation. They tended to have longer response latencies than trigeminal border units, but similar response thresholds. Many units, including those with high thresholds, were very senstive to moving stimuli, including movements of the eyelids. Very few units responded to thermal stimulation of the cornea.
To reveal central nervous system (CNS) structures involved in the baroreceptor reflex we studied the distribution of Fos protein-like immunoreactivity in the rat brain after one hour of electrical stimulation of the aortic depressor nerve (ADN). In 13 male Wistar rats under urethane the ADN was cut on both sides and the central ends were placed on stimulating electrodes. Intermittent (11 s on, 6 s off) electrical stimulation at parameters set to elicit a drop in mean arterial pressure of 15-30 mmHg was applied to one, both or neither ADNs for 1 h. CNS sections were incubated for 48 h in anti-Fos antibody and prepared for visualization of the reaction product using the ABC immunoperoxidase technique. Label was found in several discrete brain nuclei primarily on the side ipsilateral to the side of stimulation. In the medulla labelled nuclei were found in the nucleus tractus solitarius, area postrema, rostral and caudal ventrolateral medulla, nucleus ambiguus and medullary reticular formation. In the pons labelled neurons were found in the lateral and ventrolateral parabrachial nucleus, locus coeruleus, pontine reticular field and A5 region. In the forebrain labelled nuclei were observed in the peri- and paraventricular hypothalamus, supraoptic nucleus, subfornical organ, preoptic area, central nucleus of the amygdala, median preoptic area, horizontal limb of the diagonal band, bed nucleus of the stria terminalis and islands of Calleja. In control animals moderate amounts of label were present in the supraoptic nucleus and periventricular hypothalamus bilaterally. These results define central pathways involved in mediating the baroreceptor reflex.
Trigeminal nociceptive neurons within the subnucleus reticularis ventralis medullae oblongatae (SRV), which lies ventral to the trigeminal subnucleus caudalis and subnucleus reticularis dorsalis medullae oblongatae, were studied in urethane/chloralose-anesthetized cats and monkeys. These neurons were called 'SRV neurons'. They were almost regularly excited by pressure to the ipsilateral cornea or to both corneas at a strength well above the human corneal pain threshold. Most of them were activated by noxious mechanical stimulation of the pinna, face and/or tongue. A significant fraction of SRV units was responsive to tapping of the ipsilateral dorsum of the nose and/or electrical stimulation of tooth pulp afferents. Evidence was obtained that responses to tapping of the dorsum of the nose were due to mechanical distortion of the nasal mucosa. Intracellular injection of HRP into SRV neurons demonstrated that injected neurons were large neurons characteristic of the SRV. Trigeminal tractotomy just rostral to the obex did not eliminate responses of SRV units to trigeminal inputs. Neurons relaying trigeminal inputs to SRV neurons were electrophysiologically identified in the nucleus reticularis parvocellularis which is ventromedially adjacent to the subnuclei oralis and interpolaris of the trigeminal spinal tract nucleus. These findings were supported by HRP injection into the SRV. Units having receptive fields similar to those of SRV neurons were found in lamina VII of the first cervical cord, suggesting that SRV neurons may be trigeminal lamina VII neurons.
Spinal afferents to the medullary dorsal reticular nucleus were studied using the following retrograde tracers: horseradish peroxidase (diluted in dimethylsulfoxide), wheat germ agglutinin conjugated with horseradish peroxidase, and cholera toxin subunit B. Spinal cord cells projecting to that medullary region were located predominantly in medial lamina I and lamina X. Cell labelling was moderate in the medial part of laminae II-IV and sparse throughout laminae V-VII. Labelling was predominantly ipsilateral in the dorsal horn and bilateral in laminae VII and X. After mechanical lesions of the dorsal white matter which severed most of the ipsilateral cuneate fasciculus, the numbers of superficial dorsal horn cells that were labelled from the dorsal reticular nucleus were considerably decreased caudal to the lesion, which suggests that their axons utilize mostly the cuneate fasciculus. Since the medullary dorsal reticular nucleus of the rat has a predominant population of nociceptive specific neurons, it is suggested that this spino-dorsomedullary reticular pathway is involved in pain processing.
The projections of the superficial dorsal horn to the lateral reticular nucleus of the medulla oblongata of the rat, and the morphological types of spinal cord lamina I neurons involved were studied after injecting the retrograde tracer cholera toxin subunit B in the caudal portion of the lateral reticular nucleus. Only injection sites located in the lateral part of the lateral reticular nucleus caused retrograde cell labelling in the superficial dorsal horn (laminae I-III). However, injection sites covering the lateral half of the lateral reticular nucleus and the region intermediate between its lateral border and the ventrocaudal tip of the trigeminal spinal nucleus also labelled cells in the neck of the dorsal horn. In contrast, injection sites confined to the intermediate region gave rise to an almost exclusive cell labelling in laminae I-III. Because the lateral part of the lateral reticular nucleus and the adjoining lateral region are rich in noradrenergic cells, it is suggested that these may be the specific targets of laminae I-III neurons. On the basis of the solid dendritic filling achieved, labelled lamina I cells were classified structurally. Most were fusiform cells (80%) and a minority pyramidal or flattened cells (10% each). Since fusiform cells also project selectively to the parabrachial nuclei, which together with the lateral reticular nucleus have been implicated in respiratory and cardiovascular reflexes, it is suggested that this cell type may convey nociceptive input originating autonomic responses. The pyramidal cells project also in large numbers to the mesencephalic periaqueductal gray which, like the lateral reticular nucleus, exerts descending inhibition on the dorsal horn nociceptive neurons. This suggests that this cell type may activate the spinal-midbrain pain modulatory loops centred on both nuclei.
Corneal units in the trigeminal subnucleus caudalis and adjacent bulbar lateral reticular formation were studied in urethane-chloralose anesthetized cats. Corneal units were categorized into four classes: low-threshold corneal (LTC) units, high-threshold corneal (HTC) units, wide dynamic range (WDR) units with corneal input, and subnucleus reticularis ventralis (SRV) units with corneal input. Corneal receptive fields of these four classes of corneal afferent units consisted of 3-6 spots. Mechanical thresholds of LTC units were lower than 30 mg (2.6 g/mm2) and were comparable to the sensory threshold of the human cornea measured in patients with cataract. Mechanical thresholds of the other 3 classes of corneal afferent units were well above the pain threshold in the human cornea. LTC units were located in the magnocellular layer of trigeminal subnucleus caudalis and were intermingled with cutaneous low-threshold mechanoreceptive units. HTC units were coexistent with nociceptive specific units in the marginal layer and in the outer zone of substantia gelatinosa. WDR units with corneal input were found in the lateral part of trigeminal lamina V equivalent, which corresponds to the lateral part of subnucleus reticularis dorsalis. These 3 classes of corneal units were found at a level 2.7-3.5 mm caudal to the obex. SRV units were found in the dorsolateral part of SRV along the entire length of the medulla oblongata caudal to the obex. These results support the suggestion that either nonpainful sensation or pain can be evoked from the cornea.
Small amounts of the retrograde tracer WGA-apoHRP-Au complex were injected in the caudal medulla to study the spinal afferents to the subnucleus reticularis dorsalis (SRD). Labelled neurones were found at all levels of the spinal cord: the highest numbers were in the ipsilateral cervical spinal cord (mainly laminae I, V, VI, VII, VIII and X), the lowest were at the thoracic and lumbar levels bilaterally, while an intermediate density was found bilaterally at the sacral level. When injection sites were located in the underlying subnucleus reticularis ventralis (SRV), labelling was bilateral and mainly in the deep layers of the cervical spinal cord. Together with our previous electrophysiological and anatomical data, this study suggests that the SRD provides a link in spino-reticulo-spinal loops implicated in the processing of pain.
The protooncogene c-fos is expressed rapidly, transiently and polysynaptically within neurons in response to synaptic activation and voltage-gated calcium entry into the cell. The nuclear protein product of this gene (Fos) is detectable immunohistochemically 20-90 min after cell activation and remains within the nucleus for hours after expression. The present study was undertaken to identify cells within the rat medulla oblongata that express Fos-like protein in response to stimulation of afferent fibers of the carotid sinus nerve (CSN). Direct electrical stimulation of the CSN in anesthetized animals or hypoxic stimulation in either anesthetized or awake animals resulted in a consistent and discrete distribution of Fos-like immunoreactivity (Fos-LI). Fos-LI was observed bilaterally within nucleus tractus solitarius (NTS) and the ventrolateral medulla (VLM), within area postrema and nucleus raphe pallidus, and bilaterally along the ventral medullary surface. Unstimulated animals were devoid of Fos-LI within the medulla oblongata. Furthermore, neither the surgical preparations alone nor the effects of anesthesia could account for the extent of Fos-LI observed. We believe these cells represent second- and higher-order neurons within the baroreceptor and chemoreceptor reflex pathways.
C-fos is a proto-oncogene that is expressed within some neurons following depolarization. The protein product, c-fos protein, can be identified by immunohistochemical techniques. Therefore, c-fos expression might be used as a marker for neuronal activity throughout the neuraxis following peripheral stimulation. This study has analyzed patterns of c-fos expression in both control and anesthetized animals and in anesthetized rats subjected to various forms of peripheral stimulation. Labeled cells were counted in the spinal cord, brainstem, hypothalamus, and thalamus. Little c-fos immunoreactivity was found in control animals. Prolonged inhalational anesthesia increased the number of labeled cells at several brainstem sites. Noxious stimulation of anesthetized rats induced c-fos within the neuraxis in patterns consistent with data obtained from electrophysiological studies and in additional locations for which few direct electrophysiological data are available, such as the ventrolateral medulla, the posterior hypothalamic nucleus, and the reuniens and paraventricular thalamic nuclei. Gentle mechanical stimulation was ineffective in inducing c-fos-like protein. The data suggest that c-fos can be used as a transynaptic marker for neuronal activity following noxious stimulation. However, c-fos is expressed only in some kinds of neurons following peripheral stimulation, and it therefore may be an incomplete marker for nociresponsive activity. In addition, at least a few neurons express c-fos protein in the absence of noxious stimulation. Experiments analyzing c-fos expression must be designed with care, as both extraneous stimuli and anesthetic depth influence the results.
A modification of the formalin test for assessing pain and analgesia in the orofacial region of the rat is described. A formalin solution (5%) was subcutaneously injected into the upper lip, then the length of time the animal spent rubbing the injected zone was recorded. Two distinct periods of intensive rubbing activity were identified: an early phase between 0 and 3 min after the injection and a late phase between 18 and 42 min after the injection. Acetylsalicylic acid, paracetamol and morphine all had an antinociceptive effect during the two phases although incomplete during the early phase. Our results indicate that this orofacial formalin test is a valid technique for the study of orofacial pain.
In Nembutal-anesthetized rats, 31 physiologically identified medullary dorsal horn (MDH) cells were labeled with horseradish peroxidase (HRP). Ten responded only to deflection of one or more vibrissae. Six cells were activated by guard hair movement only, six by deflection of guard hairs or vibrissa(e), and seven by pinch of facial skin with serrated forceps. Different classes of low-threshold cells could not be distinguished on the basis of their somadendritic morphologies or laminar distribution. Neurons activated by multiple vibrissae were unique, however, in that one sent its axon into the medial lemniscus, and three projected into the trigeminal spinal tract. None of the guard hair-only or vibrissae-plus-guard hair neurons had such projections. Cells that responded best to noxious stimulation were located mainly in laminae I, II, and deep V, while neurons activated by vibrissa(e) and/or guard hair deflection were located in layers III, IV, and superficial V. Low-threshold neurons generally had fairly thick dendrites with few spines, whereas high-threshold cells tended to have thinner dendrites with numerous spines. Moreover, the dendritic arbors of low-threshold cells were, for the most part, denser than those of the noxious cells. Neurons with mandibular receptive fields were located in the dorsomedial portion of the MDH; cells with ophthalmic fields were found in the ventrolateral MDH, and maxillary cells were interposed. Cells sensitive to deflection of dorsal mystacial vibrissae and/or guard hairs were located ventral to those activated by more ventral hairs. Neurons with rostral receptive fields were found in the rostral MDH, while cells activated by hairs of the caudal mystacial pad, periauricular, and periorbital regions were located in the caudal MDH. Receptive-field types were encountered that have not been reported for trigeminal primary afferent neurons: multiple vibrissae; vibrissae plus guard hairs; and wide dynamic range. The latter two can be explained by the convergence of different primary afferent types onto individual neurons. Our failure to find a significant relationship between dendritic area (in the transverse plane) and the number of vibrissae suggests that primary afferent convergence may not be responsible for the synthesis of the multiple vibrissae receptive field. Excitatory connections between MDH neurons may, therefore, account for multiple vibrissae receptive fields in the MDH.
The extracellular activity of single neurons was recorded in subnucleus caudalis (medullary dorsal horn) of chloralose-anesthetized cats to test the effects of electrical and natural stimuli that activated afferents supplying the jaw and tongue muscles as well as the face, teeth, and intraoral mucosa. Many caudalis neurons that could be functionally classified on the basis of their cutaneous receptive-field properties as low-threshold mechanoreceptive (LTM), wide-dynamic-range (WDR), or nociceptive-specific (NS) neurons could be excited by muscle afferent stimuli. Only five neurons were encountered that received muscle afferent inputs and had no demonstrable cutaneous, dental, or mucosal input. The muscle afferent inputs were a particular feature of the cutaneous nociceptive (i.e., WDR and NS) neurons. Approximately two-thirds of this nociceptive neuronal population (n = 109) could be excited by jaw and/or tongue muscle stimulation, whereas only a small proportion of the LTM neuronal population (n = 247) was activated by muscle afferent stimulation. Neurons with a demonstrated direct axonal projection to the contralateral thalamus as well as nonprojection neurons received muscle afferent inputs. The caudalis nociceptive neurons receiving muscle as well as cutaneous afferent inputs had receptive-field properties comparable to those previously described for caudalis cutaneous nociceptive neurons; they were predominantly located in laminae I/II and V/VI, and many also received convergence of tooth pulp afferent inputs. These neurons generally had larger cutaneous receptive fields than neurons unresponsive to muscle afferent stimulation. The muscle afferent inputs were considered to be predominantly of a nociceptive character for several reasons. These included the long latency and high threshold of most neuronal responses evoked by electrical stimulation of the muscle afferents, the predominance of afferents of small diameter in some of the muscle nerves stimulated, the preferential responsiveness to the muscle afferent stimulation of neurons that were functionally identified as cutaneous nociceptive neurons, and the responsiveness of most of the neurons excited by electrical stimulation of the muscle nerves also to noxious mechanical or thermal stimulation of muscle and the injection of two or more algesic chemicals into small arteries supplying the jaw and tongue muscles. Of the algesic chemicals used in this study (7% NaCl, KCl, bradykinin, histamine, 5-HT), the first two were found to be the most effective and to cause the most rapidly induced excitation.(ABSTRACT TRUNCATED AT 400 WORDS)
The face of the lightly anesthetized macaque was stimulated with noxious and non-noxious natural stimuli and with brief electrical pulses. Unit responses were recorded extracellularly from somatosensory cells located in bulbar lateral reticular formation (LRF) regions which are ventromedially contiguous with the trigeminal nucleus caudalis. The following functional properties characterized the cells of the LRF. (1) They had a wide range of thresholds, latencies and stimulus following capacities. (2) They most frequently adapted slowly to stimulation although a variety of tonic and phasic response patterns was also observed. (3) They exhibited inhibitory as well as excitatory modes of action. (4) They did not have a clearly defined somatotopy. (5) They were innervated by projections from fields which varied considerably in size. (6) They revealed marked differences in modality and sensitivity in that some responded exclusively and other differentially to specific forms of painful and non-painful stimuli applied to their peripheral fields.The results closely correspond to those reported for LRF cells in other species and they support the recent proposal that the LRF has a fundamental role in the mediation of trigeminal pain operating in this capacity in conjunction with other nuclear zones of the medulla.
Discharges were recorded extracellularly with tungsten microelectrodes from single neurons that were located in the cat's caudal bulbar reticular formation in anesthetized or decerebrated preparations. The responses to qualitatively and quantitatively graded mechanical and thermal stimuli were analyzed with respect to the lowest stimulus intensity needed to elicit a response, the response pattern evoked by the adequate stimulus, the receptive field size and the position of the receptive field on the body surface. The most typical neuron found in both types of preparations had an excitatory response when a heavy-noxious somatic stimulus was applied to a receptive field that was restricted to the face. The cutaneous fields were large and in some cases extended over the entire head. The qualitative classification of a heavy-noxious stimulus was confirmed by a close correlation between thermal and pressure thresholds of the units and psychophysical evaluations of various stimulus intensities. It was concluded that the caudal medullary reticular formation was physiologically distinguishable from more rostral parts of the brain stem reticular formation because of the functional specificity of a large proportion of the units in this part of the reticular formation.
The activity of 160 single neurons excited by electrical stimulation of the canine tooth pulp was studied in the subnucleus caudalis (medullary dorsal horn) and the subnucleus oralis of the trigeminal (V) spinal tract nucleus in chloralose-anesthetized cats to test the effects of natural as well as electrical stimulation of the tooth pulp. The neurons were functionally classified on the basis of their cutaneous receptive-field properties as low-threshold mechanoreceptive (LTM), wide dynamic range (WDR), or nociceptive specific (NS). The orofacial receptive-field properties and responses evoked by electrical stimulation of the tooth pulp indicated that the oralis and caudalis neurons examined had characteristics typical of those previously documented for oralis LTM neurons and for caudalis LTM, WDR, and NS neurons. Each neuron was also tested with cold and warm stimulation of the canine tooth, and some neurons were also tested for responsiveness to thermal stimulation of the premolar tooth or to mechanical and chemical stimuli delivered to the dentine of the canine tooth. Although all the neurons could be excited by electrical stimulation of the pulp, we found that the only neurons that consistently responded to thermal pulp stimuli were those located in the V subnucleus caudalis. Moreover, only those caudalis neurons that had been functionally classified as nociceptive (4 WDR and 21 NS neurons) showed this responsiveness. Heating of the canine or premolar tooth excited 24 of these 25 nociceptive neurons; cooling activated only 3, and none of the small number of neurons tested with mechanical and chemical stimulation of the dentine was excited. The response of the nociceptive neurons to heating of the tooth contrasted with the responses of the same neurons to pinching and heating of their cutaneous receptive field.(ABSTRACT TRUNCATED AT 400 WORDS)
1. The medulla oblongata caudal to the obex was explored with glass capillary microelectrodes filled with fast green dye in urethan-chloralose-anesthetized cats. Concurrently, the trigeminal integument was mechanically stimulated. Two classes of units, which were activated by innocuous mechanical stimulation of the ipsilateral trigeminal integument, were identified. Both of them showed a somatotopic organization. 2. Units maximally activated by tactile stimulation of a small receptive field in the ipsilateral trigeminal integument were located in pars magnocellularis of the trigeminal subnucleus caudalis. Receptive field falling within, the mandibular distribution were found in the dorsomedial part of pars magnocellularis, maxillary fields occurred next, and ophthalmic fields were represented ventrolaterally. The rostrum (the mouth and nose) was represented just behind the obex, and the receptive field represented gradually shifted to the periphery of the face along the rostrocaudal axis, finally proceeding to dermatomes of cervical segments. At each transverse plane, tactile sensation of the most rostral segment of the represented integument projected onto the marginal zone of pars magnocellularis immediately adjacent to substantia gelatinosa, while the most peripheral segment projected onto the core of quasi-semicircular pars magnocellularis. The result is in contrast with the scheme proposed by previous investigators. 3. Within the lateral part of subnucleus reticularis dorsalis medullae oblongatae, which is ventromedially contiguous with pars magnocellularis, trigeminal units homologous with lamina V units in the spinal dorsal horn were identified. Typically, these units had a graded response in the center of the receptive field, but responded only to strong mechanical stimuli applied to the periphery of the receptive field. In the center of the receptive field, tactile stimulation of a small restricted area was effective, and the tactile receptive field showed a somatotopic organization. The mandibular distribution was represented dorsomedially, the maxillary distribution next to that, and the ophthalmic distribution laterally. Furthermore, a rostrocaudal differentiation similar to that observed with tactile units in pars magnocellularis was found. Hence, a double somatotopic representation of tactile sensation within the caudal medulla oblongata was indicated.
The responses of 475 neurons to orthodromic oral-facial stimuli were studied in adult cats anesthetized with chloralose. These neurons were located in trigeminal (V) subnucleus caudalis, which has recently been termed the medullary dorsal horn in view of its functional and structural similarities with the spinal cord dorsal horn. The oral-facial stimuli included nonnoxious tactile and occasionally thermal stimuli, tooth pulp stimuli, noxious mechanical and radiant-heat stimuli, and graded electrical stimulation of several oral-facial nerves, skin, and mucosa. In addition, antidromic stimulation techniques were employed to determine if the functionally identified neurons projected to the posterior thalamus or V subnucleus oralis. All the units could be broadly classified as either cutaneous nociceptive or low-threshold mechanoreceptive (LTM) neurons. A proportion of neurons in each of these two categories also responded to tooth pulp stimulation and were considered in a separate category of pulp-driven neurons. A total of 304 LTM neurons in layers III/IV, V, and VI of caudalis were studied. Most responded exclusively to low-threshold oral-facial tactile stimuli (85 also responded to tooth pulp stimuli). The short-latency (<5 ms) excitatory responses to peripheral electrical stimuli noted for most of these neurons indicated an A-β-afferent input, but some had longer latency discharges suggestive of an A-δ input. Approximately 20% of the LTM neurons also had visceral afferent convergence since they also responded to glossopharyngeal nerve (IX) or superior laryngeal nerve (SLN) stimulation. Almost 12% had a direct axonal projection to the thalamus, with a mean antidromic conduction velocity of 20.4 m/s. In addition, 50% of the LTM neurons tested with stimulation of the subnucleus oralis were found to project to oralis, but with a significantly slower conduction velocity (mean 5.7 m/s) than the trigeminothalamic axons. The 122 cutaneous nociceptive neurons examined were located in a somatotopically organized fashion in layers I-II and also in layers V-VI. Approximately 15% projected directly to the thalamus and 33% had a slower conducting projection to oralis; the conduction velocities of their axons projecting to the thalamus or oralis were significantly slower than those of LTM neurons. The projection from caudalis nociceptive (and LTM) neurons to oralis may be involved in the recently demonstrated ascending modulating influence that caudalis exerts over synaptic transmission in oralis neurons. Most nociceptive neurons had oral-facial mechanoreceptive fields limited to one V division or less, and approximately 15% showed visceral afferent convergence from IX and SLN. They were classified as either wide dynamic range (WDR) or nociceptive specific (NS). The WDR neurons responded to nonnoxious stimuli as well as to noxious mechanical and thermal stimuli. Although they had significantly shorter response latencies to peripheral electrical stimuli than did NS neurons, their responses indicated excitatory inputs from A-δ, and in some cases from C-afferents, as well as from A-β-afferent fibers. Some of the WDR neurons with a C-fiber input showed the phenomenon of 'windup.' One group of NS neurons responded to heavy pressure and pinch, and most also responded to noxious radiant heat. They had electrically evoked responses indicative of excitatory inputs from A-δ afferents, and approximately 50% also showed C-fiber discharges. Other NS neurons responded to pinch but not to heavy pressure, and most of these also responded to noxious radiant heat. Their electrically evoked responses suggested excitatory inputs only from A-δ-afferents. One-third of the caudalis population of cutaneous nociceptive and LTM neurons could be activated by electrical stimulation of the maxillary and/or mandibular canine tooth pulp. Nearly all of these neurons also had cutaneous or oral afferent inputs. Some pulp-driven neurons were found to project to the thalamus or V subnucleus oralis, were located in layers I-II, III/IV, V, and VI of caudalis, and had the cutaneous or mucosal receptive-field properties described above for nociceptive or LTM neurons. Approximately 25% of the neurons classified as LTM had a pulp afferent input, as did 12% of WDR neurons and almost 50% of the NS neurons. The response properties of these neurons are compared with those of other pulp-driven neurons previously described in the V main sensory nucleus and subnucleus oralis. Although it would be premature to rule out the involvement in dental pain of these rostral V brain stem neurons, our results indicate that neurons in caudalis, and particularly the pulp-driven NS neurons, may be of critical importance for the appreciation of dental pulp pain.
The objectives of the present study were 1) to utilize Fos immunohistochemistry as a marker for neuronal activity in order to examine the population of neurons in the medulla that is engaged by activation of nociceptive peripheral afferents and 2) to determine whether catecholamine-containing neurons in the medulla also express noxious heat-evoked Fos-like immunoreactivity. Noxious heating of the hindpaw evoked specific patterns of Fos-like immunoreactivity in the medulla in regions known to be involved in both nociceptive processing and cardiovascular regulation. Noxious heating of the hindpaw significantly increased the mean number of neurons expressing Fos-like immunoreactivity in the contralateral ventrolateral medulla. Increased numbers of Fos-positive neurons also were observed in both the ipsilateral and the contralateral A1 catecholamine cell groups. Similarly, in the contralateral medullary dorsal reticular fields, noxious heating of the hindpaw significantly increased the mean number of neurons expressing Fos-like immunoreactivity. In contrast, in the paramedian reticular nucleus, noxious heating of the hindpaw resulted in a significant decrease in the mean number of neurons expressing Fos-like immunoreactivity. No significant differences in the mean numbers of neurons expressing Fos-like immunoreactivity were noted in the A2, C1, or C2/C3 medullary catecholamine cell groups. These results suggest that noxious stimuli affect pools of neurons in the medulla with multiple physiological functions.
An ultrastructural study is made of the synaptic contacts occurring between structures labelled anterogradely and retrogradely in the superficial dorsal horn following injections of cholera toxin subunit B or horseradish peroxidase in the dorsal reticular nucleus of the medulla oblongata of the rat. Both tracers revealed labelled axonal boutons in lamina I with round synaptic vesicles and a few large granular vesicles making asymmetrical synaptic contacts upon labelled somata and dendrites. After injections of Phaseolus vulgaris leucoagglutinin in the dorsal reticular nucleus, labelled boutons identical to those revealed by the two other tracers were presynaptic to unlabelled somata and dendrites. In addition, dorsoreticular neurons were labelled retrogradely following injections of cholera toxin subunit B into the superficial dorsal horn of the cervical enlargement. These observations show the occurrence of a reciprocal connection between dorsal reticular and lamina I neurons. Considering the putative excitatory nature of the axodendritic contacts in lamina I, a positive feedback circuit is suggested, whereby the nociceptive signals transmitted to the dorsal medullary reticular formation by marginal neurons are intensified.
Corneal input to the spinal trigeminal nucleus (Vsp) was assessed by examining Fos-like immunoreactivity (Fos-LI) after chemical irritant stimulation by mustard oil in chloralose-anesthetized cats. The distribution of Fos-LI within the ipsilateral Vsp was bimodal: a dominant group of cells within the superficial laminae at caudal levels of subnucleus caudalis and a second group of cells within the ventrolateral pole of Vsp at obex levels and within the adjacent interstitial islands. Few Fos-positive cells were seen within the Vsp rostral to the mid-portion of subnucleus interpolaris or within the contralateral Vsp. To assess the involvement of caudal portions of the Vsp in mediating the adrenal and autonomic responses to corneal stimulation, mustard oil was applied before and after lidocaine blockade of the Vsp at obex levels in a second group of cats. Corneal stimulation alone increased significantly (P < 0.001) the adrenal secretion of catecholamines, adrenal blood flow, mean arterial pressure and heart rate. With the exception of heart rate, the adrenal and autonomic responses to mustard oil were greatly attenuated or abolished by lidocaine blockade of the ipsilateral Vsp at the level of the obex, a region that displayed a high number of Fos-positive cells after corneal stimulation. These results indicate that neurons within the Vsp at or more caudal than the level of the obex process chemical irritant input from the cornea and are necessary for corneal-evoked changes in adrenal and autonomic function.
Brainstem catecholaminergic neurons have been implicated as mediating adaptive autonomic and neuroendocrine responses to cardiovascular challenges. To clarify the nature of this involvement, immuno- and hybridization histochemical methods were used to follow c-fos expression in these neurons in response to acute stimuli that differentially affect blood pressure and volume. From low basal levels, hypotensive hemorrhage (15%) provoked a progressive increase in the number and distribution of Fos-immunoreactive (ir) nuclei in the nucleus of the solitary tract (NTS), the A1 and C1 cell groups of the ventrolateral medulla, and in the pontine A5, locus coeruleus, and lateral parabrachial cell groups peaking at 2.0-2.5 hours after the challenge. Fos-ir ventrolateral medullary neurons, subsets of which were identified as projecting to the paraventricular hypothalamic nucleus or spinal cord, were predominantly aminergic, whereas most of those in the NTS were not. Infusion of an angiotensin II antagonist blunted hemorrhage-induced Fos expression in the area postrema, and attenuated that seen elsewhere in the medulla and pons. Nitroprusside-induced isovolemic hypotension yielded a pattern of c-fos induction similar to that seen following hemorrhage, except in the area postrema and the A1 cell group, where the response was muted or lacking. Phenylephrine-induced hypertension stimulated a restricted pattern of c-fos expression, largely limited to induced hypertension stimulated a restricted pattern of c-fos expression, largely limited to non-aminergic neurons, whose distribution in the NTS conformed to the termination patterns of primary baroreceptor afferents, and in the ventrolateral medulla overlapped in part with those of vagal cardiomotor and depressor neurons. These findings underscore the importance of brainstem catecholaminergic neurons in effecting integrated homeostatic responses to cardiovascular challenges and their ability to responding strategically to specific modalities of cardiovascular information. They also foster testable predictions as to effector neuron populations that might be recruited to respond to perturbations in individual circulatory parameters.
The medulla oblongata was examined with Fos and tyrosine hydroxylase (TH) immunohistochemistry after 2 h of intermittent nasopharyngeal stimulation with formaldehyde vapour in the conscious rabbit. The stimulation caused apnoea, bradycardia and a rise in blood pressure known to be associated with vigorous vasoconstriction. Fos-positive neurons occurred in the spinal trigeminal nucleus, the nucleus tractus solitarius, the raphe nuclei and the ventrolateral medulla. In the rostral ventrolateral medulla, 68% of the Fos-positive neurons were TH-positive C1 cells. Our data indicate that nasopharyngeally-evoked peripheral vasoconstriction is associated with activation of C1 neurons.
Horseradish peroxidase (HRP) was stereotaxically injected into the nucleus accumbens (Acb), and visceral noxious stimulation given by injecting formalin into the stomach. Sections of the medulla were subjected to HRP reaction combined with immunohistochemical reactions for Fos protein (ABC method) and tyrosine hydroxylase (TH, PAP method). The catecholaminergic neurons of the medulla (including vagal complex, ventrolateral medulla and reticular formation between them) which expressed Fos protein and projected to Acb were studied. The results showed that HRP retrogradely labeled cells were seen in the medulla bilaterally with apparent ipsilateral predominance and TH-LI and Fos-LI single labeled cells were bilaterally distributed; HRP/TH,TH/Fos double labeled neurons were more numerous than HRP/Fos double-labeled neurons. HRP/TH/Fos triple-labeled neurons were small in number and were mainly distributed in the nucleus tractus solitarii (nTS) and ventrolateral medulla (VLM), but only a few labeled cells were located in RF between nTS and VLM. It is concluded that TH-LI neurons in the medulla projected to Acb and some of them expressed Fos protein after noxious stimulation of the stomach.
The location of neurons in the caudal medulla oblongata that project to the superficial or deep dorsal horn was studied in the rat, by means of retrograde labelling from confined spinal injection sites. The tracer cholera toxin subunit B was injected into laminae I-III (fuve rats) or I-V (three rats) at C4-7 spinal segments. Neurons projecting to the superficial dorsal horn were located in the dorsomedial part of the dorsal reticular nucleus ipsilaterally, the subnucleus commissuralis of the nucleus tractus solitarius bilaterally, and a region occupying the lateralmost part of the ventrolateral reticular formation between the lateral reticular nucleus and the caudal pole of the spinal trigeminal nucleus, pars caudalis, bilaterally. Neurons projecting to the deep dorsal horn, which were only labelled when laminae I-V were filled by the tracer, occurred in the dorsomedial and ventrolateral parts of the dorsal reticular nucleus and in the ventral reticular nucleus bilaterally. A few cells were located in the above described lateralmost portion of the ventrolateral reticular formation bilaterally and in the ventral portion of the ipsilateral cuneate nucleus. In the light of previous data demonstrating that dorsal horn neurons project to the dorsal reticular nucleus, the ventrolateral reticular formation, and the nucleus tractus solitarius, and that neurons in these three medullary regions are involved in pain inhibition at the spinal level, the descending projections demonstrated here suggest the occurrence of spino-medullary-spinal loops mediating the analgesic actions elicited in each nucleus upon the arrival of nociceptive input from the dorsal horn.
We have studied the induction of c-fos immunoreactivity (c-fos IR) in catecholaminergic and vasopressinergic immunoreactive neurons after repeated phosphate-buffered saline (PBS) loading or after repeated elicitation of the baroreceptor reflex via repeated infusion of the vasoconstrictor agent L-phenylephrine. About 75% and 30%, respectively, of the tyrosine-hydroxylase immunoreactive (IR) cell bodies of the ventral noradrenaline (NA) A1/adrenaline (A) C1 and dorsal NA A2/A C2 areas and 60% and 30%, respectively, of the phenylethanolamine N-methyltransferase IR nerve cells of the adrenaline C1 and C2 areas and 25% of the vasopressin (VP) IR neurons of the supraoptic (SO) nucleus developed nuclear c-fos IR after repeated PBS loading. This phenomenon remained unaltered by the repeated elicitation of the baroreceptor reflex. These results suggest that the activation of volume receptors promotes homeostatic responses via activation of early genes in subsets of central medullary noradrenaline and adrenaline neurons and SO VP neurons of the urethane-anaesthetized rat.
We have used the evoked expression of the immediate early gene-encoded proteins (c-Fos, Fos B, Jun B, Jun D, c-Jun and Krox-24) to monitor sensory processing in the hindbrain structures of rats undergoing somatic inflammation. Experiments were performed on freely moving animals that did not experience constraints other than those imposed by the disease itself. Local injections of chemicals were used to cause subcutaneous inflammation of the plantar foot or monoarthritis by intracapsular injection. Labelling was studied at survival times that corresponded either to the time points of maximum labelling in the spinal cord (4 h for the subcutaneous model, 24 h and two weeks for the monoarthritis model) or at survival times that corresponded to the chronic phase of monoarthritis evolution (six, nine and 15 weeks). Controls consisted of freely moving, unstimulated animals. Basal expression was observed for all immediate early genes and in a variety of stuctures, but always remained moderate. All immediate early gene-encoded protein expressions except c-Jun were evoked, but except for c-Fos, and to a lesser extent Jun D, intensities of staining always remained faint. The following results will be mainly based on c-Fos expression, as this protein proved to be the most effective marker for all the survival times studied.
Antibodies against the c-fos protein product Fos were used to map the first- and higher-order neurons in the rabbit medulla oblongata after electrical stimulation of the vagus nerve. Fos immunoreactivity appeared bilaterally except in the nucleus tractus solitarii. Seven areas were labeled: the nucleus tractus solitarii, the area postrema, the subnucleus lateralis caudalis magnocellularis medullar oblongata, the lateral reticular nucleus, the ambiguus nucleus, the dorsal part of the spinal trigeminal nucleus, the nucleus reticularis lateralis, the lateral border of the external cuneatus nucleus, the medial part of the inferior olivary nucleus (subnucleus beta). The last two areas have never been visualized with conventional tracing techniques and may represent higher-order neurons connected to visceral vagal pathways. No labeling was observed in the nodose ganglion.
Noxious somatic stimuli elicit vasopressin secretion, an effect thought to result from activation of a facilitatory input from Al catecholamine cells of the medulla oblongata. To better characterize the Al cell response and effects on other neuroendocrine Al projection targets, particularly within the paraventricular nucleus, we have now mapped c-fos expression in neurochemically identified catecholamine and neurosecretory cells following a noxious somatic stimulus. Unilateral hind paw pinch significantly increased c-fos expression in contralateral Al cells whereas other brainstem catecholamine cell groups were unaffected. Expression of c-fos was also increased in the supraoptic nucleus, this effect being more pronounced for vasopressin than oxytocin neurosecretory cells and, as with Al cells, primarily on the side contralateral to the stimulated paw. In contrast, the increase in the paraventricular nucleus was greater in oxytocin rather than in vasopressin cells. Additionally there was a significant rise in c-fos expression in medial parvocellular paraventricular nucleus cells of noxiously stimulated animals. Notably, the majority of tuberoinfundibular corticotropin-releasing factor cells are located in this medial parvocellular zone.
The nociceptive nature of the neurons of the superficial dorsal horn (laminae I-III) which project to the medullary ventrolateral reticular formation is studied in the rat. Medullary injections of Fluoro-Gold showed exclusive retrograde labeling of laminae I-III cells when the tracer filled a zone intermediate between the lateral tip of the lateral reticular nucleus and the spinal trigeminal nucleus, pars caudalis. This zone is here called VLMlat. Following noxious mechanical or thermal stimulation of the skin, double-labeled neurons, which stained retrogradely and were Fos-immunoreactive, prevailed in laminae I and IIo. Double-labeled neurons were few in lamina IIi after thermal stimulation and entirely lacking in lamina III after the two kinds of stimulation. Findings in lamina I confirm previous electrophysiological data (see Menétrey et al., J. Neurophysiol., 52 (1984) 595-611) showing that lamina I cells projecting to the ventrolateral reticular medulla convey noxious messages. The occurrence of numerous double-labeled cells in lamina IIo suggests that this lamina is also involved in nociceptive transmission to the VLMlat.
Recent studies have provided evidence that excitatory amino acid antagonists can exert analgesic effects in animals. These studies, however, have focused primarily on phasic pain or hyperalgesia rather than tonic pain. The present study evaluates the effects of systemic administration of Memantine (1-amino-3,5-dimethyl-adamantane), a clinically used N-methyl-D-aspartate (NMDA) receptor antagonist, on formalin-induced phasic and tonic pain behavior in the rat. Memantine (2.5, 5.0, 10.0 and 20.0 mg/kg) or normal saline was injected i.p. 1 h prior to a s.c. injection of formalin (5%, 50 microliters) into the vibrissal pad of adult rats (n = 5/group). Pain behavior was measured by the number of seconds of formalin-induced face grooming during a 42-min post-injection observation period. Saline-injected animals displayed a biphasic face-grooming response, consisting of an early, phasic phase (0-6 min) and a delayed, prolonged tonic phase (12-42 min). Memantine at doses of 2.5-10 mg/kg produced a significant dose-related inhibition of the second phase (65-93%) and a much smaller inhibition of the first phase (up to 52%). A higher dose (20 mg/kg) further inhibited both phases but also produced other motor effects (increased exploratory and decreased freezing behavior, hind-paw weakness and gait ataxia) which were not observed at the lower doses. These results suggest that the NMDA receptor antagonist Memantine can block formalin-induced tonic and, to a lesser extent, phasic pain, at doses that do not alter observed motor behaviors.
We have used the evoked expression of the immediate early gene-encoded proteins (Krox-24, c-Fos, Fos B, Jun D, Jun B, c-Jun) to monitor visceral processing in both the spinal cord and hindbrain structures of rats undergoing either mechanical colorectal or chemical intraperitoneal stimulation. Experiments were conducted under controlled volatile anaesthesia to suppress affective reactions that visceral stimulations may induce. The results refer to the effects of anaesthesia alone, and of both innocuous and noxious stimulations.
C-fos expression in hypothalamic
- Dw Smith
- Day
- Ta
Smith DW, Day TA (1994) C-fos expression in hypothalamic
Distri-bution and neurochemical phenotypes of caudal medullary neurons activated to express cFos following peripheral admin-istration of cholecystokinin Somatic sensory proper-ties of bulbar reticular neurons
- Rinaman L Jg
- Em Stricker
- Hoffman
- Jp
- T Takenaka
- Encabo
Rinaman L, Verbalis JG, Stricker EM, Hoffman GE (1993) Distri-bution and neurochemical phenotypes of caudal medullary neurons activated to express cFos following peripheral admin-istration of cholecystokinin. J Comp Neurol 338:475-490 S egundo JP, Takenaka T, Encabo H (1967) Somatic sensory proper-ties of bulbar reticular neurons. J Neurophysiol 30:1221-1238