Medullary Dorsal Reticular Nucleus Research Articles

Sex-related correspondence between mechanical hypersensitivity and the discharge of medullary pain control neurons in neuropathic rats

Here we studied whether the sex-related difference in mechanical hypersensitivity induced by neuropathy is associated with the discharge rate of medullary pain control neurons. We performed experiments in male and female rats with spared nerve injury (SNI) model of peripheral neuropathy. Mechanical hypersensitivity was assessed behaviorally by monofilaments. Discharge rates of pain-control neurons were determined using in vivo single unit recordings under light anesthesia. Recording targets were two medullary nuclei involved in descending pain control: the rostral ventromedial medulla (RVM) and the medullary dorsal reticular nucleus (DRt). Based on the response to peripheral noxious stimulus, neurons were classified as pronociceptive RVM ON-like or DRt neurons, or antinociceptive RVM OFF-like neurons. Behavioral results indicated that the mechanical hypersensitivity induced by SNI was significantly stronger in females than males. The ongoing discharge rates of pronociceptive RVM ON-like neurons were higher and those of antinociceptive RVM OFF-like neurons lower in SNI females than SNI males. Ongoing discharge rates of pronociceptive DRt neurons were not significantly different between SNI females and males. The results suggest that a sex difference in the discharge rate of pain control neurons in the RVM but not DRt may contribute to the maintenance of stronger neuropathic hypersensitivity in females.

Monoaminergic and Opioidergic Modulation of Brainstem Circuits: New Insights Into the Clinical Challenges of Pain Treatment?

The treatment of neuropathic pain remains a clinical challenge. Analgesic drugs and antidepressants are frequently ineffective, and opioids may induce side effects, including hyperalgesia. Recent results on brainstem pain modulatory circuits may explain those clinical challenges. The dual action of noradrenergic (NA) modulation was demonstrated in animal models of neuropathic pain. Besides the well-established antinociception due to spinal effects, the NA system may induce pronociception by directly acting on brainstem pain modulatory circuits, namely, at the locus coeruleus (LC) and medullary dorsal reticular nucleus (DRt). The serotoninergic system also has a dual action depending on the targeted spinal receptor, with an exacerbated activity of the excitatory 5-hydroxytryptamine 3 (5-HT3) receptors in neuropathic pain models. Opioids are involved in the modulation of descending modulatory circuits. During neuropathic pain, the opioidergic modulation of brainstem pain control areas is altered, with the release of enhanced local opioids along with reduced expression and desensitization of μ-opioid receptors (MOR). In the DRt, the installation of neuropathic pain increases the levels of enkephalins (ENKs) and induces desensitization of MOR, which may enhance descending facilitation (DF) from the DRt and impact the efficacy of exogenous opioids. On the whole, the data discussed in this review indicate the high plasticity of brainstem pain control circuits involving monoaminergic and opioidergic control. The data from studies of these neurochemical systems in neuropathic models indicate the importance of designing drugs that target multiple neurochemical systems, namely, maximizing the antinociceptive effects of antidepressants that inhibit the reuptake of serotonin and noradrenaline and preventing desensitization and tolerance of MOR at the brainstem.

The Modulation of Pain by Metabotropic Glutamate Receptors 7 and 8 in the Dorsal Striatum.

The dorsal striatum, apart from controlling voluntary movement, displays a recently demonstrated pain inhibition. It is connected to the descending pain modulatory system and in particular to the rostral ventromedial medulla through the medullary dorsal reticular nucleus. Diseases of the basal ganglia, such as Parkinson's disease, in addition to being characterized by motor disorders, are associated with pain and hyperactivation of the excitatory transmission. A way to counteract glutamatergic hyperactivation is through the activation of group III metabotropic glutamate receptors (mGluRs), which are located on presynaptic terminals inhibiting neurotransmitter release. So far the mGluRs of group III have been the least investigated, owing to a lack of selective tools. More recently, selective ligands for each mGluR of group III, in particular positive and negative allosteric modulators, have been developed and the role of each subtype is starting to emerge. The neuroprotective potential of group III mGluRs in pathological conditions, such as those characterized by elevate glutamate, has been recently shown. In the dorsal striatum, mGluR7 and mGluR8 are located at glutamatergic corticostriatal terminals and their stimulation inhibits pain in pathological conditions such as neuropathic pain. The two receptors in the dorsal striatum have instead a different role in pain control in normal conditions. This review will discuss recent results focusing on the contribution of mGluR7 and mGluR8 in the dorsal striatal control of pain. The role of mGluR4, whose antiparkinsonian activity is widely reported, will also be addressed.

The medullary dorsal reticular nucleus as a relay for descending pronociception induced by the mGluR5 in the rat infralimbic cortex

Metabotropic glutamate receptor 5 (mGluR5) activation in the infralimbic cortex (IL) induces pronociceptive behavior in healthy and monoarthritic rats. Here we studied whether the medullary dorsal reticular nucleus (DRt) and the spinal TRPV1 are mediating the IL/mGluR5-induced spinal pronociception and whether the facilitation of pain behavior is correlated with changes in spinal dorsal horn neuron activity. For drug administrations, all animals had a cannula in the IL as well as a cannula in the DRt or an intrathecal catheter. Heat-evoked paw withdrawal was used to assess pain behavior in awake animals. Spontaneous and heat-evoked discharge rates of single DRt neurons or spinal dorsal horn wide-dynamic range (WDR) and nociceptive-specific (NS) neurons were evaluated in lightly anesthetized animals.Activation of the IL/mGluR5 facilitated nociceptive behavior in both healthy and monoarthritic animals, and this effect was blocked by lidocaine or GABA receptor agonists in the DRt. IL/mGluR5 activation increased spontaneous and heat–evoked DRt discharge rates in healthy but not monoarthritic rats. In the spinal dorsal horn, IL/mGluR5 activation increased spontaneous activity of WDR neurons in healthy animals only, whereas heat-evoked responses of WDR and NS neurons were increased in both experimental groups. Intrathecally administered TRPV1 antagonist prevented the IL/mGluR5-induced pronociception in both healthy and monoarthritic rats.The results suggest that the DRt is involved in relaying the IL/mGluR5-induced spinal pronociception in healthy control but not monoarthritic animals. Spinally, the IL/mGluR5-induced behavioral heat hyperalgesia is mediated by TRPV1 and associated with facilitated heat-evoked responses of WDR and NS neurons.

Galanin-Mediated Behavioural Hyperalgesia from the Dorsomedial Nucleus of the Hypothalamus Involves Two Independent Descending Pronociceptive Pathways.

Activation of the dorsomedial nucleus of the hypothalamus (DMH) by galanin (GAL) induces behavioural hyperalgesia. Since DMH neurones do not project directly to the spinal cord, we hypothesized that the medullary dorsal reticular nucleus (DRt), a pronociceptive region projecting to the spinal dorsal horn (SDH) and/or the serotoninergic raphe-spinal pathway acting on the spinal 5-HT3 receptor (5HT3R) could relay descending nociceptive facilitation induced by GAL in the DMH. Heat-evoked paw-withdrawal latency (PWL) and activity of SDH neurones were assessed in monoarthritic (ARTH) and control (SHAM) animals after pharmacological manipulations of the DMH, DRt and spinal cord. The results showed that GAL in the DMH and glutamate in the DRt lead to behavioural hyperalgesia in both SHAM and ARTH animals, which is accompanied particularly by an increase in heat-evoked responses of wide-dynamic range neurons, a group of nociceptive SDH neurones. Facilitation of pain behaviour induced by GAL in the DMH was reversed by lidocaine in the DRt and by ondansetron, a 5HT3R antagonist, in the spinal cord. However, the hyperalgesia induced by glutamate in the DRt was not blocked by spinal ondansetron. In addition, in ARTH but not SHAM animals PWL was increased after lidocaine in the DRt and ondansetron in the spinal cord. Our data demonstrate that GAL in the DMH activates two independent descending facilitatory pathways: (i) one relays in the DRt and (ii) the other one involves 5-HT neurones acting on spinal 5HT3Rs. In experimental ARTH, the tonic pain-facilitatory action is increased in both of these descending pathways.

Role of glutamate receptors in the dorsal reticular nucleus in formalin‐induced secondary allodynia

The role of glutamate receptors present in the medullary dorsal reticular nucleus (DRt) in the formalin test and formalin-induced secondary nociception was studied in rats. Secondary mechanical allodynia was assessed with von Frey filaments applied to the rat's hindpaw, and secondary thermal hyperalgesia was evaluated with the tail-immersion test. The selective glutamate receptor antagonists MK801 (N-methyl-D-aspartate receptor antagonist), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (AMPA/KA receptor antagonist) and A841720 (metabotropic glutamate 1 receptor antagonist) were injected into the DRt before or 6 days after formalin injection in the rat. In the formalin test, the three antagonists significantly reduced the number of flinches in both phases of the test. DRt microinjection of MK801 or A841720, but not of CNQX, reduced both secondary nociceptive behaviors. Moreover, pre-treatment with the three antagonists injected into the DRt prevented the development of secondary mechanical allodynia and secondary thermal hyperalgesia. Similarly, in these rats, the number of c-Fos-like immunoreactive neurons were markedly reduced in both the superficial and deep lamina of the dorsal horn. Our findings support the role of DRt as a pain facilitator in acute and chronic pain states, and suggest a key role of glutamate receptors during the development and maintenance of formalin-induced secondary allodynia.

The Striatum and Pain Modulation

The aim of this review was to give a general aspect of the sensorial function of the striatum related to pain modulation, which was intensively studied in our laboratory. We analyse the effect of electrical and chemical stimulation of the striatum on the orofacial pain, especially that produced by tooth pulp stimulation of the lower incisors. We demonstrated specific sites within the nucleus which electrical or chemical stimulation produced inhibition of the nociceptive jaw opening reflex. This analgesic action of the striatum was mediated by activation of its dopamine D(2) receptors and transmitted through the indirect pathways of the basal ganglia and the medullary dorsal reticular nucleus (RVM) to the sensorial nuclei of the trigeminal nerve. Its mechanism of action was by inhibition of the nociceptive response of the second order neurons of the nucleus caudalis of the V par.

Parathyroid hormone 2 receptor and its endogenous ligand tuberoinfundibular peptide of 39 residues are concentrated in endocrine, viscerosensory and auditory brain regions in macaque and human

Parathyroid hormone receptor 2 (PTH2R) and its ligand, tuberoinfundibular peptide of 39 residues (TIP39) constitute a neuromodulator system implicated in endocrine and nociceptive regulation. We now describe the presence and distribution of the PTH2R and TIP39 in the brain of primates using a range of tissues and ages from macaque and human brain. In situ hybridization histochemistry of TIP39 mRNA, studied in young macaque brain, due to its possible decline beyond late postnatal ages, was present only in the thalamic subparafascicular area and the pontine medial paralemniscal nucleus. In contrast, in situ hybridization histochemistry in macaque identified high levels of PTH2R expression in the central amygdaloid nucleus, medial preoptic area, hypothalamic paraventricular and periventricular nuclei, medial geniculate, and the pontine tegmentum. PTH2R mRNA was also detected in several human brain areas by RT-PCR. The distribution of PTH2R-immunoreactive fibers in human, determined by immunocytochemistry, was similar to that in rodents, including dense fiber networks in the medial preoptic area, hypothalamic paraventricular, periventricular and infundibular (arcuate) nuclei, lateral hypothalamic area, median eminence, thalamic paraventricular nucleus, periaqueductal gray, lateral parabrachial nucleus, nucleus of the solitary tract, sensory trigeminal nuclei, medullary dorsal reticular nucleus, and dorsal horn of the spinal cord. Co-localization suggested that PTH2R fibers are glutamatergic, and that TIP39 may directly influence hypophysiotropic somatostatin containing and indirectly influence corticotropin releasing-hormone containing neurons. The results demonstrate that TIP39 and the PTH2R are expressed in the brain of primates in locations that suggest involvement in regulation of fear, anxiety, reproductive behaviors, release of pituitary hormones, and nociception.

Parathyroid hormone 2 receptor and its endogenous ligand TIP39 in the brain of macaque and human

Event Abstract Back to Event Parathyroid hormone 2 receptor and its endogenous ligand TIP39 in the brain of macaque and human Attila G. Bago1, 2, Eugene Dimitrov3, Richard Saunders3, Laszlo Seress4, Miklos Palkovits2, Ted B. Usdin3 and Arpad Dobolyi2* 1 National Institute of Neurosurgery, Hungary 2 Hungarian Academy of Sciences and Semmelweis University, Department of Anatomy, Hungary 3 National Institute of Mental Health, United States 4 University of Pécs, Hungary Parathyroid hormone 2 receptor (PTH2R) and its ligand, tuberoinfundibular peptide of 39 residues (TIP39) form a neuromodulator system in rodents present in limbic, endocrine, auditory, and viscerosensory brain areas. In this study, we describe the presence and distribution of PTH2R and TIP39 in the primate brain. RT-PCR experiments revealed PTH2R mRNA expression in several areas and cell groups of the human brain. Subsequent in situ hybridization histochemistry in macaque brain confirmed high levels of PTH2R expression in the central amygdaloid nucleus, the medial preoptic area, the hypothalamic paraventricular and periventricular nuclei, the medial nucleus of the medial geniculate body and the pontine tegmentum. Fluorescent amplification immunochemistry studies in humans demonstrated high density of PTH2R-immunoreactive fibers in many brain regions including the medial preoptic area, the paraventricular hypothalamic nucleus, the median eminence, the paraventricular nucleus of thalamus, the periaqueductal gray, the parabrachial nuclei, the nucleus of the solitary tract, the spinal trigeminal nucleus, the medullary dorsal reticular nucleus, and the dorsal horn of the spinal cord. However, no immunolabeled cell bodies were found suggesting that PTH2R is promptly transported from cell bodies towards axon terminals. In contrast to the widespread distribution of PTH2R, TIP39 mRNA was detectable only in the subparafascicular area of the posterior thalamus and the medial paralemniscal nucleus in the lateral pons. The results demonstrate that TIP39 and the PTH2R are expressed in the brain of primates with locations suggesting involvement in regulation of fear, anxiety, reproductive behaviors, release of pituitary hormones, and nociception. Support was provided by the Hungarian Science Foundation NKTH-OTKA K67646 grant for AD, the consortial FP6 EU Grant BrainNet II LSHM-CT-2004-503039 for MP, and the NIMH Intramural Research Program for TBU. Arpád Dobolyi is grantee of the Bolyai János Scholarship. Conference: 12th Meeting of the Hungarian Neuroscience Society, Budapest, Hungary, 22 Jan - 24 Jan, 2009. Presentation Type: Poster Presentation Topic: Homeostatic regulatory mechanisms Citation: Bago AG, Dimitrov E, Saunders R, Seress L, Palkovits M, Usdin TB and Dobolyi A (2009). Parathyroid hormone 2 receptor and its endogenous ligand TIP39 in the brain of macaque and human. Front. Syst. Neurosci. Conference Abstract: 12th Meeting of the Hungarian Neuroscience Society. doi: 10.3389/conf.neuro.01.2009.04.001 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 02 Mar 2009; Published Online: 02 Mar 2009. * Correspondence: Arpad Dobolyi, Hungarian Academy of Sciences and Semmelweis University, Department of Anatomy, Budapest, Hungary, dobolyi@ana.sote.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Attila G Bago Eugene Dimitrov Richard Saunders Laszlo Seress Miklos Palkovits Ted B Usdin Arpad Dobolyi Google Attila G Bago Eugene Dimitrov Richard Saunders Laszlo Seress Miklos Palkovits Ted B Usdin Arpad Dobolyi Google Scholar Attila G Bago Eugene Dimitrov Richard Saunders Laszlo Seress Miklos Palkovits Ted B Usdin Arpad Dobolyi PubMed Attila G Bago Eugene Dimitrov Richard Saunders Laszlo Seress Miklos Palkovits Ted B Usdin Arpad Dobolyi Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Dynamic of migration of HSV‐1 from a medullary pronociceptive centre: antinociception by overexpression of the preproenkephalin transgene

Herpes Simplex Virus type 1 (HSV-1) vectors are known to inhibit nociceptive transmission at the spinal cord after peripheral applications. Similar approaches may also be useful when applied at the supraspinal pain control system as the system includes pronociceptive (facilitatory) components. We performed a study aimed to analyse the migration of HSV-1 along with the inhibition of pronociception from the medullary dorsal reticular nucleus (DRt), a major facilitatory component of the supraspinal pain control system. To study the migration, a HSV-1 vector expressing lacZ under control of the human cytomegalovirus (hCMV) promoter was injected in the DRt and the expression of beta-galactosidase (beta-gal) was detected at 2, 4, 7, 10 and 14 days. Numerous beta-gal-immunoreactive neurons were observed at the injection site until day 4, and at some of the brain areas projecting to the DRt until day 7. To block the pronociceptive effects of the DRt, a HSV-1 vector expressing the preproenkephalin transgene, under the control of hCMV promoter, was injected into the DRt. Behavioural evaluation was performed at the time-points referred above, using the paw withdrawal latency test to evaluate thermal nociceptive responses. Anti-hyperalgesic effects persisted during 4 days, decreasing after that time-point. The present study demonstrates that selective migration of HSV-1 should be considered in gene therapy strategies based on HSV-1 injections into the brain. The study also shows that it is possible to decrease pain facilitation from the brain using opioidergic inhibition of pronociceptive supraspinal areas.

Contribution by DRt descending facilitatory pathways to maintenance of spinal neuron sensitization in rats

We investigated in different experimental rat models the potential facilitatory contribution of the medullary dorsal reticular nucleus (DRt) descending pathway to the expressions of the sensory spinal neuron sensitization such as increased spontaneous and noxious evoked activities, responsivity to heterotopic afferences stimulation and long lasting afterdischarges (ADs). We carried out experiments by recording from ipsilateral lumbar Wide Dynamic Range (WDR) neurons and by simultaneously monitoring the DRt neuron activity in neuropathic pain rats with chronic constriction injury of one sciatic nerve (CCI), in sham-operated and in “intact” rats. In particular, we recorded the spinal neuron spontaneous activities and the activities evoked by noxious stimulations of ipsi- and contralateral sciatic supplied areas before and during DRt activity blockade. Additionally, in “intact rats” we modulated WDR activity by iontophoretic NMDA to mimic CCI WDR hyperactivity without peripheral damage. We found that during DRt activity blockade in CCI rat neurons and in “intact” rat NMDA-treated neurons, the spontaneous activity was significantly reduced, the responses to contralateral sciatic area stimulation were reduced or suppressed, the responses to ipsilateral sciatic area were poorly affected (slightly reduced or unaffected), except for the poststimulus afterdischarges that were mostly suppressed. In sham-operated rats, the neuronal activity was not affected by DRt blockade. The finding that during the DRt nucleus blockade some expressions of spinal neurons sensitization, seemingly associated to sensory disorders in neuropathic pain, fade or extinguish designates a likely facilitatory role of DRt in the maintenance of neuronal sensitization and thus a contribution to neuropathic pain state.

Novel applications of common stereology software to represent the complete distribution, density and spatial organization of anterogradely labelled fibers in neuroanatomical tract-tracing studies

In a recent study we have adapted for the first time available software used in spatial neuronal organization studies to represent the complete fiber distribution of a brain projection [Leite-Almeida H, Valle-Fernandes A, Almeida A. Brain projections from the medullary dorsal reticular nucleus: an anterograde and retrograde tracing study in the rat. Neuroscience 2006;140:577–95]. Here we describe the technique in detail using the injection of biotinylated-dextran amine (BDA) in the rat cuneate nucleus and its projection through the pyramidal decussation as an example. Camera-lucida-like computer drawings were produced with StereoInvestigator software. Using high magnification lens, rat brain sections were screened and lines superimposed with virtually all fibers and terminal boutons present in each brain section. Additionally, in each screening field of the section, all focal planes were inspected and lines depicted only on focused fibers and/or fiber segments. Later, NeuroExplorer software was used to interpret fiber depth information and obtain the three-dimensional pathway of each fiber bundle along the rostrocaudal extension of each section. This computer methodology presents several advantages over classical camera-lucida hand-made drawings: (i) the software is prepared to work interchangeably in different magnifications without misplacements, which allows the representation of virtually all labelled fibers and, therefore, results in a much more accurate fiber location in each brain section; (ii) three-dimensional information of all individual fibers and fiber bundles is recorded; (iii) the method is more flexible allowing, for example, to colour differently labelled fibers according to their profiles (e.g. terminal or ramified fiber arborizations and terminal boutons).

Brain projections from the medullary dorsal reticular nucleus: An anterograde and retrograde tracing study in the rat

In the last 15 years a role has been ascribed for the medullary dorsal reticular nucleus as a supraspinal pain modulating area. The medullary dorsal reticular nucleus is reciprocally connected with the spinal dorsal horn, is populated mainly by nociceptive neurons and regulates spinal nociceptive processing. Here we analyze the distribution of brain projections from the medullary dorsal reticular nucleus using the iontophoretic administration of the anterograde tracer biotinylated-dextran amine and the retrograde tracer cholera toxin subunit B.Fibers and terminal boutons labeled from the medullary dorsal reticular nucleus were located predominately in the brainstem, although extending also to the forebrain. In the medulla oblongata, anterograde labeling was observed in the orofacial motor nuclei, inferior olive, caudal ventrolateral medulla, rostral ventromedial medulla, nucleus tractus solitarius and most of the reticular formation. Labeling at the pons-cerebellum level was present in the locus coeruleus, A5 and A7 noradrenergic cell groups, parabrachial and deep cerebellar nuclei, whereas in the mesencephalon it was located in the periaqueductal gray matter, deep mesencephalic, oculomotor and anterior pretectal nuclei, and substantia nigra. In the diencephalon, fibers and terminal boutons were found mainly in the parafascicular, ventromedial, and posterior thalamic nuclei and in the arcuate, lateral, posterior, peri- and paraventricular hypothalamic areas. Telencephalic labeling was consistent but less intense and concentrated in the septal nuclei, globus pallidus and amygdala.The well-known role of the medullary dorsal reticular nucleus in nociception and its pattern of brain projections in rats suggests that the nucleus is possibly implicated in the modulation of: (i) the ascending nociceptive transmission involved in the motivational-affective dimension of pain; (ii) the endogenous supraspinal pain control system centered in the periaqueductal gray matter–rostral ventromedial medulla–spinal cord circuitry; (iii) the motor reactions associated with pain.

The medullary dorsal reticular nucleus enhances the responsiveness of spinal nociceptive neurons to peripheral stimulation in the rat.

Single-unit spinal recordings combined with application of glutamate into the medullary dorsal reticular nucleus were used to assess the action of this nucleus upon deep dorsal horn neurons in rats. Injection of high glutamate concentrations (10 and 100 mm) induced a dramatic and long-lasting increase of the responses of wide-dynamic range neurons to electrical stimulation of the sciatic nerve in the noxious range, without affecting ongoing discharges. Post-stimulus time histograms revealed that this increase concerned the post-discharge, but not A- or C-fibre-mediated responses, which remained unchanged independently of the stimulation frequency applied. The onset of the glutamate-induced response enhancement occurred with a concentration-dependent time delay and developed slowly until its maximum. These data indicate that the medullary dorsal reticular nucleus exerts a facilitating action upon deep dorsal horn wide-dynamic range neurons by enhancing their capacity to respond to peripheral stimulation through prolongation of their discharge. This action is accompanied by the strengthening of wind-up of deep dorsal horn wide-dynamic range neurons, hence providing a plausible substrate for chronic pain states. These results are in agreement with previous behavioural studies suggesting a pronociceptive role for the dorsal reticular nucleus [Almeida et al. (1996) Brain Res. Bull., 39, 7-15; Almeida et al. (1999) Eur. J. Neurosci., 11, 110-122], and support the involvement of a reverberating circuit, previously described in morphological studies [Almeida et al. (1993) Neuroscience, 55, 1093-1106; Almeida et al. (2000) Eur. J. Pain, 4, 373-387], which probably operates only at a certain threshold of activation.

Brain afferents to the medullary dorsal reticular nucleus: a retrograde and anterograde tracing study in the rat.

The medullary dorsal reticular nucleus (DRt) was recently shown to belong to the supraspinal pain control system; neurons within this nucleus give origin to a descending projection that increases spinal nociceptive transmission and facilitates pain perception [Almeida et al. (1999), Eur. J. Neurosci., 11, 110-122]. In the present study, the areas of the brain that may modulate the activity of DRt neurons were investigated by using of tract-tracing techniques. Injection of a retrograde tracer into the DRt resulted in labelling in multiple areas of the brain. In the contralateral orbital, prelimbic, infralimbic, insular, motor and somatosensory cortices labelling was prominent, but a smaller ipsilateral projection from these same areas was also detected. Strong labelling was also noted in the central amygdaloid nucleus, bed nucleus of stria terminalis and substantia innominata. Labelled diencephalic areas were mainly confined to the hypothalamus, namely its lateral and posterior areas as well as the paraventricular nucleus. In the mesencephalon, the periaqueductal grey, red nucleus and deep mesencephalic nucleus were strongly labelled, whereas, in the brainstem, the parabrachial nuclei, rostroventromedial medulla, nucleus tractus solitarius, spinal trigeminal nucleus, and the parvocellular, dorsal, lateral and ventral reticular nuclei were the most densely labelled regions. All deep cerebellar nuclei were labelled bilaterally. These data suggest that the DRt integrates information from the somatosensory, antinociceptive, autonomic, limbic, pyramidal and extrapyramidal systems while triggering its descending facilitating action upon the spinal nociceptive transmission.

Reciprocal connections between the medullary dorsal reticular nucleus and the spinal dorsal horn in the rat

The synaptic architecture of spinal afferents of the dorsal portion (DRtd) of the medullary dorsal reticular nucleus (DRt) is studied. After iontophoretic injections of cholera toxin subunit B (CTb) into the superficial (laminae I–II), deep (laminae IV–V) or entire (laminae I–V) dorso-ventral extent of the spinal dorsal horn at the cervico-thoracic or lumbo-sacral levels, axonal boutons of two distinct types were labelled in the DRtd. Type A boutons (82% after cervico-thoracic injections and 92% after lumbo-sacral injections) were roundish, small and presented few mitochondria and small, round synaptic vesicles. Type B boutons (18% after cervico-thoracic injections and 8% after lumbo-sacral injections) were elongated, two to three times larger, and exhibited numerous mitochondria and larger round vesicles. Both types of bouton established asymmetrical synaptic contacts with small dendritic profiles and, less frequently, with dendritic trunks and perikarya. Retrograde labelling occurred in the postsynaptic profile of 15–18% type A boutons labelled from any injection site. Taken together with previous data on DRt-spinal synaptic contacts at the superficial dorsal horn, the present results point to the occurrence of a reciprocal excitatory loop connecting the dorsal DRt and lamina I, which may be at the basis of the DRt-mediated pain-facilitating effects described recently.

The medullary dorsal reticular nucleus facilitates pain behaviour induced by formalin in the rat.

The influence of the dorsal reticular nucleus (DRt) on pain behaviour during the formalin test was studied in the rat by lesioning the nucleus through local application of electrical current or quinolinic acid. Animals in which the DRt was lesioned ipsilaterally to the paw injected with formalin spent less time in focused (licking, biting or scratching the injected paw) and total (focused pain behaviour plus protection of the injected paw during movements) pain behaviour, and showed paw-jerks less frequently than non-lesioned animals in both phases 1 and 2 of the test. Animals in which the DRt was lesioned contralaterally to the injected paw presented a decrease in total pain behaviour and number of paw-jerks only during phase 2. The number of superficial (laminae I-II) and deep (laminae III-VI) spinal dorsal horn cells expressing the c-fos proto-oncogene 2 h after subcutaneous injection of formalin was reduced by 34% and 50%, respectively, in animals with an ipsilateral DRt lesion as compared to non-lesioned rats. No differences in c-fos expression were observed after lesioning the DRt contralateral to the formalin injection. The results indicate that the DRt is involved in the facilitation of nociception during the formalin test by enhancing the response capacity of dorsal horn neurons to noxious stimulation. It is suggested that the pronociceptive action of the DRt is mediated by the reciprocal connections it establishes with the spinal dorsal horn.

Activation by cutaneous or visceral noxious stimulation of spinal neurons projecting to the medullary dorsal reticular nucleus in the rat: a c-fos study.

The involvement of spinal neurons in the transmission of cutaneous and visceral nociceptive input to the medullary dorsal reticular nucleus was studied. Rats were injected with cholera toxin subunit B in the left dorsal reticular nucleus and subjected 4 days later to noxious mechanical, thermal or chemical stimulation of the proximal internal aspect of the left thigh, or to chemical stimulation of the urinary bladder. Sections of spinal segments T13-L3 were processed immunocytochemically for cholera toxin subunit B and Fos protein. The percentage of double-labelled cells in the population of Fos-positive cells was higher in lamina I (1-4%) than in deeper laminae (0-0.7%) following all stimuli. The percentage of double-labelled cells in the population of retrogradely labelled cells was 30-53% in lamina I and 0-5% in laminae III-X. Visceral stimulation activated more retrogradely labelled lamina I cells than any kind of cutaneous stimulation. Pyramidal cells were activated in higher numbers than multipolar and flattened cells after thermal cutaneous or visceral stimulation, and in lower numbers than multipolar cells after mechanical stimulation. These results suggest that, in the experimental conditions used, spinal cord cells conveying noxious input to the dorsal reticular nucleus are concentrated in lamina I. They further indicate that the spinal-dorsal reticular nucleus pathway plays a major role in the transmission of nociceptive visceral input, and point to the preferential involvement of pyramidal cells in cutaneous thermal and visceral processing.

Projection sites of superficial or deep dorsal horn in the dorsal reticular nucleus.

The termination sites in the medullary dorsal reticular nucleus (DRt) of axons arising from superficial or deep dorsal horn cells were analysed by anterograde tracing with biotinylated dextran (BD). Following BD injections in spinal laminae I-III or in these laminae plus the dorsal funiculus, labelled fibres and boutons were located in the dorsal most ipsilateral portion of the DRt. Injections covering laminae III-V or laminae I-V resulted in bilateral labelling of the ventral DRt and ipsilateral labelling of the dorsal DRt. These findings, together with previous data concerning the origin of DRt projections to the superficial or deep dorsal horn, suggest the occurrence of reciprocal anatomical loops connecting the dorsal or the ventral areas of both regions.

Descending projections from the medullary dorsal reticular nucleus make synaptic contacts with spinal cord lamina I cells projecting to that nucleus: An electron microscopic tracer study in the rat

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.