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Convergent data showed that bulbo-spinal serotonergic projections exert complex modulatory influences on nociceptive signaling within the dorsal horn. These neurons are located in the B3 area which comprises the median raphe magnus (RMg) and the lateral paragigantocellular reticular (LPGi) nuclei. Because LPGi 5-HT neurons differ from RMg 5-HT neurons regarding both their respective electrophysiological properties and responses to noxious stimuli, we used anatomical approaches for further characterization of the respective spinal projections of LPGi versus RMg 5-HT neuron subgroups. Adult Sprague-Dawley rats were stereotaxically injected into the RMg or the LPGi with the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L). The precise location of injection sites and RMg vs LPGi spinal projections into the different dorsal horn laminae were visualized by PHA-L immunolabeling. Double immunofluorescent labeling of PHA-L and the serotonin transporter (5-HTT) allowed detection of serotonergic fibers among bulbo-spinal projections. Anterograde tracing showed that RMg neurons project preferentially into the deep laminae V-VI whereas LPGi neuron projections are confined to the superficial laminae I-II of the ipsilateral dorsal horn. All along the spinal cord, double-labeled PHA-L/5-HTT immunoreactive fibers, which represent only 5–15% of all PHA-L-immunoreactive projections, exhibit the same differential locations depending on their origin in the RMg versus the LPGi. The clear-cut distinction between dorsal horn laminae receiving bulbo-spinal serotonergic projections from the RMg versus the LPGi provides further anatomical support to the idea that the descending serotonergic pathways issued from these two bulbar nuclei might exert different modulatory influences on the spinal relay of pain signaling neuronal pathways., Highlights • Anterograde tracing and 5-HT transporter immunochemistry allowed investigations of bulbo-spinal 5-HT projections in rats. • 5-HT fibers represent ∼5% vs ∼15% of descending projections from n. raphe magnus vs n. paragigantocellularis, respectively. • 5-HT fibers from the n. raphe magnus project mainly into dorsal horn laminae V-VI, and to lower extent into laminae I-II. • In contrast, 5-HT fibers from the n. paragigantocellularis project densely and exclusively into superficial laminae I-II. • These anatomical data suggest that spinal 5-HT from raphe magnus vs paragigantocellularis n. exert distinct pain controls.

La TGIR Huma-Num a labellisé pour quatre ans (2014-2017) un consortium consacré à la 3D pour réaffirmer, au cœur des projets 3D, les enjeux scientifiques des SHS. Le consortium 3D SHS s’est proposé de diffuser à travers cet ouvrage des guides de bonnes pratiques (archivage 3D, logiciels et matériels, vocabulaire, cahier des charges ) à destination de la communauté SHS.

Serotonin (5-HT) neurons are involved in wake promotion and exert a strong inhibitory influence on rapid eye movement (REM) sleep. Such effects have been ascribed, at least in part to the action of 5-HT at post-synaptic 5-HT1A receptors (5-HT1AR) in the brainstem, a major wake/REM sleep regulatory center. However, the neuroanatomical substrate through which 5-HT1AR influence sleep remains elusive. We therefore investigated whether a brainstem structure containing a high density of 5-HT1AR mRNA, the GABAergic Gudden's dorsal tegmental nucleus (DTg), may contribute to 5-HT-mediated regulatory mechanisms of sleep-wake stages. We first found that bilateral lesions of the DTg promote wake at the expense of sleep. In addition, using local microinjections into the DTg in freely moving mice, we showed that local activation of 5-HT1AR by the prototypical agonist 8-OH-DPAT enhances wake and reduces deeply REM sleep duration. The specific involvement of 5-HT1AR in the latter effects was further demonstrated by ex vivo extracellular recordings showing that the selective 5-HT1AR antagonist WAY 100635 prevented DTg neuron inhibition by 8-OH-DPAT. We next found that GABAergic neurons of the ventral DTg exclusively targets glutamatergic neurons of the lateral mammillary nucleus (LM) in the posterior hypothalamus by means of anterograde and retrograde tracing techniques using cre driver mouse lines and a modified rabies virus. Altogether, our findings strongly support the idea that 5-HT-driven enhancement of wake results from 5-HT1AR-mediated inhibition of DTg GABAergic neurons that would in turn disinhibit glutamatergic neurons in the mammillary bodies. We therefore propose a Raphe→DTg→LM pathway as a novel regulatory circuit underlying 5-HT modulation of arousal., SCOPUS: ar.j, info:eu-repo/semantics/published

Cree en 2014 par le conseil scientifique de la tres grande infrastructure de recherche (Tgir) Huma-Num, le consortium 3D coordonne par Archeovision federe dix equipes de recherche qui developpent des technologies et produisent des modeles 3D dans le cadre de projets de recherche en sciences humaines et sociales. Le consortium 3D fait partie de la derniere vague de labellisation d’Huma-Num et rejoint ainsi ArchiPolis, Archives des Ethnologues, Archives des Mondes Contemporains, Cahier, Cartes ...

The present study was designed to record electrophysiological responses to graded noxious thermal stimuli of serotonergic and nonserotonergic neurons in the lateral paragigantocellular reticular (LPGi) and the raphe magnus (RMg) nuclei in rats. All of the neurons recorded were juxtacellularly filled with neurobiotin and identified with double immunofluorescent labeling for both neurobiotin and serotonin. Under halothane anesthesia (0.75%), noxious thermal stimuli ⩾48°C activated almost all (88%) of the serotonergic neurons located within the LPGi (n=16). The increase in firing was clear (3.4±0.3spike/s: mean of responses above the population median) and sustained during the whole application of strong thermal noxious stimuli, with a high mean threshold (48.3±0.3°C) and large receptive fields. Recording of serotonergic neurons in the RMg (n=21) demonstrated that the proportion of strongly activated (>2spike/s) neurons (19% vs 59% for the LPGi) as well as the magnitude of the activation (2.1±0.4spike/s: mean of responses above the population median) to thermal noxious stimuli were significantly lower than in the LPGi (P

Among receptors mediating serotonin actions in pain control, the 5-HT(7)R is of special interest because it is expressed by primary afferent fibers and intrinsic GABAergic and opioidergic interneurons within the spinal dorsal horn. Herein, we investigated whether GABA and/or opioids contribute to 5-HT(7)R-mediated control of neuropathic pain caused by nerve ligation. Acute administration of 5-HT(7)R agonists (AS-19, MSD-5a, E-55888) was found to markedly reduce mechanical and thermal hyperalgesia in rats with unilateral constriction injury to the sciatic nerve (CCI-SN). In contrast, mechanical hypersensitivity caused by unilateral constriction injury to the infraorbital nerve was essentially unaffected by these ligands. Further characterization of the anti-hyperalgesic effect of 5-HT(7)R activation by the selective agonist E-55888 showed that it was associated with a decrease in IL-1ß mRNA overexpression in ipsilateral L4-L6 dorsal root ganglia and lumbar dorsal horn in CCI-SN rats. In addition, E-55888 diminished CCI-SN-associated increase in c-Fos immunolabeling in superficial laminae of the lumbar dorsal horn and the locus coeruleus, but increased c-Fos immunolabeling in the nucleus tractus solitarius and the parabrachial area in both control and CCI-SN rats. When injected intrathecally (i.t.), bicuculline (3 μg i.t.), but neither phaclofen (5 μg i.t.) nor naloxone (10 μg i.t.), significantly reduced the anti-hyperalgesic effects of 5-HT(7)R activation (E-55888, 10 mg/kg s.c.) in CCI-SN rats. These data support the idea that 5-HT(7)R-mediated inhibitory control of neuropathic pain is underlain by excitation of GABAergic interneurons within the dorsal horn. In addition, 5-HT(7)R activation-induced c-Fos increase in the nucleus tractus solitarius and the parabrachial area suggests that supraspinal mechanisms might also be involved.

Activités de fouilles soutenues par : Université de Lille 3 (UMR 8164 du CNRS HALMA-IPEL) avec la collaboration du Musée Oiasso d’Irun, de l’Association Arkeolan, de la Soprintendenza archeologica di Pompei et de l’École française de Rome Mis au jour entre octobre 1823 et février 1824, le sanctuaire de Fortune qui réunit un temple monumental et des annexes destinées à l’organisation du culte avec logement, cuisine et salle de banquet (VII, 4, 1 et 2) est une découverte ancienne mainte fois me...

Non-technical summary Defence reactions are physiological responses to imminent danger. They include increased blood pressure and heart rate, and a reduction in the reflex cardiac response to changes in blood pressure. Two regions in the brain, the hypothalamus and the periaqueductal grey area, known to be involved in pathological conditions such as anxiety, are involved in the production of these responses. However, there is no direct connection between those regions. In this study we report that the midbrain cuneiform nucleus links the hypothalamus and periaqueductal grey area. Our findings may explain how anxiety is related to cardiovascular pathologies. Abstract Defence responses triggered experimentally in rats by stimulation of the dorsomedial nucleus of the hypothalamus (DMH) and the dorsolateral periaqueductal grey matter (PAG) inhibit the cardiac baroreflex response (i.e. bradycardia). It has also been proposed that the midbrain cuneiform nucleus (CnF) is involved in active responses. Our aim was to identify the neurocircuitry involved in defence-induced baroreflex inhibition, with a particular focus on the link between DMH, CnF and dorsolateral PAG. Microinjection of the anterograde tracer Phaseolus vulgaris leucoaggutinin into the CnF revealed a dense projection to the dorsolateral PAG. Moreover, activation of neurons in the CnF induced increased expression of Fos protein in the dorsolateral PAG. Inhibition of neurons of the CnF or dorsolateral PAG prevented the inhibition of baroreflex bradycardia induced by DMH or CnF stimulation, respectively. These results provide a detailed description of the brain circuitry underlying acute baroreflex modulation by neurons of the DMH. Our data have shown for the first time that the CnF plays a key role in defence reaction-associated cardiovascular changes; its stimulation, from the DMH, activates the dorsolateral PAG, which, in turn, inhibits baroreflex bradycardia.

Activités de fouilles menées par : École française de Rome [en collaboration avec la surintendance archéologique de Rome] Cette quatrième campagne des fouilles menées par l’École française de Rome, dirigées par Martine Dewailly et conduites en collaboration avec Fedora Filippi de la Soprintendenza per i beni archeologici di Roma, est théoriquement la dernière à s’inscrire dans le cadre du projet financé par l’Agence Nationale de la Recherche intitulé « Du stade de Domitien à l’actuelle piazza...

Several voltage-gated sodium channels are expressed in primary sensory neurons where they control excitability and participate in the generation and propagation of action potentials. Peripheral nerve injury-induced alterations in both tetrodotoxin (TTX)-sensitive and TTX-resistant sodium channels have been proposed to contribute to neuropathic pain caused by such lesion. We herein investigated whether the blockade of TTX-sensitive channels could reduce pain-related behaviors and evoked c-Fos immunoreactivity in rats with neuropathic pain produced by chronic unilateral constriction injury to either the sciatic nerve or the infraorbital nerve. Acute as well as subchronic administration of TTX (1-6 mug/kg s.c.) was found to suppress for up to 3 h allodynia and hyperalgesia in sciatic nerve-ligated rats. In contrast, TTX was only moderately effective in rats with ligated infraorbital nerve. In sciatic nerve-ligated rats, TTX administration prevented the increased c-Fos immunoreactivity occurring in the dorsal horn of the lumbar cord and some supraspinal areas in response to light mechanical stimulation of the nerve-injured hindpaw. The anti-allodynia/antihyperalgesia caused by TTX in these neuropathic rats was promoted by combined treatment with naloxone (0.5 mg/kg s.c.) but unaffected by the 5-HT(1B) receptor antagonist F11648 (0.5 mg/kg s.c.) and the alpha(2)-adrenergic receptor antagonist idazoxan (0.5 mg/kg i.v.). In contrast, the anti-allodynic and anti-hyperalgesic effects of TTX were significantly attenuated by co-administration of morphine (3 mg/kg s.c.) or the cholecystokinin(2)-receptor antagonist CI-1015 (0.1 mg/kg i.p.). These results indicate that TTX alleviates pain-related behaviors in sciatic nerve-lesioned rats through mechanisms that involve complex interactions with opioidergic systems.

Restraint stress produces changes in the sleep pattern that are mainly characterized by a delayed increase in rapid eye movement sleep (REMS) amounts. Because the serotonin (5-HT) and the hypocretin (hcrt) systems that regulate REMS are interconnected, we used mutant mice deficient in the 5-HT transporter (5-HTT−/−) to examine the role of 5-HT and hcrt neurotransmissions in the sleep response to stress.In contrast to wild-type mice, restraint stress did not induce a delayed increase in REMS amounts in5-HTT−/−mice, indicating impaired sleep homeostasis in mutants. However, pharmacological blockade of the hcrt type 1 receptor (hcrt-R1) before restraint stress restored the REMS increase in5-HTT−/−mice. In line with this finding,5-HTT−/−mutants displayed after restraint stress higher long-lasting activation of hypothalamic preprohcrt neurons than wild-type mice and elevated levels of the hcrt-1 peptide and the hcrt-R1 mRNA in the anterior raphe area. Thus, hypocretinergic neurotransmission was enhanced by stress in5-HTT−/−mice. Furthermore, in5-HTT−/−but not wild-type mice, hypothalamic levels of the 5-HT metabolite 5-hydroxyindole acetic acid significantly increased after restraint stress, indicating a marked enhancement of serotonergic neurotransmission in mutants.Altogether, our data show that increased serotonergic -and in turn hypocretinergic- neurotransmissions exert an inhibitory influence on stress-induced delayed REMS. We propose that the direct interactions between hcrt neurons in the hypothalamus and 5-HT neurons in the anterior raphe nuclei account, at least in part, for the adaptive sleep–wakefulness regulations triggered by acute stress.

Bernard Jean-François, Dewailly Martine, Lovergne Edwige, Caldarini Carla, Méniel Patrice, Russo Jacopo. Piazza Navone, 62. In: Mélanges de l'École française de Rome. Antiquité, tome 121, n°1. 2009. Antiquité. pp. 297-314.

Our first aim was to elucidate the mechanisms underlying the hypotensive response elicited by 5-HT(2) receptor activation in the nucleus tractus solitarius (NTS). In pentobarbitone-anaesthetized rats, intra-NTS administration of 2,5-dimethoxy-4-iodoamphetamine (DOI), a wide spectrum 5-HT(2) receptor agonist, but not an antagonist of selective 5-HT(2B) and 5-HT(2C) receptors, produced a decrease in blood pressure and heart rate. The maximal cardiovascular changes obtained by DOI (0.5 pmol) could be almost completely abolished by prior intra-NTS microinjection (10 pmol) of MDL-100907, a selective 5-HT(2A) receptor antagonist, but not by 5-HT(2B) or 5-HT(2C) receptor antagonists. In addition, using extracellular recordings we found that the large majority of identified cardiovascular rostroventrolateral medulla (RVLM) neurons were almost totally inhibited by NTS 5-HT(2A) receptor stimulation. We then investigated whether intra-NTS administration of a subthreshold dose (0.05 pmol) of DOI, known to facilitate the cardiovagal component of the baroreflex, could also modulate the sympathoinhibitory component of this reflex. These experiments showed that neither the decrease in the activity of the cardiovascular RVLM neurons and lumbar sympathetic nerve activities produced by aortic occlusion (gain of the baroreflex), nor the hypotensive response elicited by aortic nerve stimulation, were potentiated by the microinjection of DOI under such conditions. These data show that activation of 5-HT(2A), but not 5-HT(2B) or 5-HT(2C), receptors, located on NTS neurons, elicits depressor and bradycardic responses, and that this 5-HT(2A)-mediated hypotension is produced via the inhibition of RVLM cardiovascular neurons. In addition, NTS 5-HT(2A) receptor activation facilitates the cardiac but not the sympathetic baroreflex response.

Despite recent discoveries made in myeloproliferative disorders other than chronic myelogenous leukemia, which it is hoped will result in earlier diagnosis, and better evaluation and management of patients, hematological evolution to myelofibrosis, acute leukemia, and myelodysplastic syndromes (AL/MDS) remain major causes of long-term mortality in polycythemia vera (PV) and essential thrombocythemia (ET) patients. Evaluation of long-term leukemogenic risk of currently available drugs, therefore, is crucial. We report updated results of three French prospective trials of hydroxyurea and pipobroman in PV and ET patients with a median follow-up longer than 10 years. The results show that the incidence of AL/MDS is higher than previously reported with no evidence of a plateau (with approximately 40% of AL/MDS cases occurring after the 12th year of follow-up). Although hydroxyurea currently remains the first choice in the treatment of high-risk PV and ET patients, the use of nonleukemogenic drugs, such as interferon alpha (IFN-alpha) or anagrelide, should be assessed more widely in randomized trials using accurate diagnostic criteria and taking into account the presence of the JAK2 mutation, given that they may have an impact on disease evolution.

An open-label, phase II non-randomised trial was conducted with darbepoetin (DAR), an erythropoiesis-stimulating factor with prolonged half-life, at a weekly dose of 300 mug subcutaneously in 62 anaemic patients with myelodysplastic syndrome (MDS) with an endogenous erythropoietin (EPO) level

This study investigated the responses of posterior triangular (PoT) thalamic neurons to tactile and noxious calibrated stimuli in anesthetized rats. We report here that 41% of PoT units responded to cutaneous stimulation, in most cases, by increasing strongly their firing. Forty-five percent of the responding units were nociceptive specific (NS), 19% were nociceptive nonspecific (NNS), and 36% were tactile. The NS units responded only to frankly noxious stimuli applied to relatively large receptive fields (several parts of the body). They encoded nociceptive temperatures chiefly in 46-50°C ranges. The NNS units resembled NS units but also responded to innocuous stimuli. Tactile units responded chiefly to repeated innocuous stimuli applied to very small receptive fields (one to two fingers or vibrissae).A representative sample of PoT somatosensory neurons, characterized first by their response to innocuous and noxious cutaneous stimuli, were filled with juxtacellular injection of biotin-dextran that made it possible to label adequately the soma, the dendrites, and the entire axon of PoT neurons. We observed that the axons of NS neurons terminated only in secondary somatosensory (S2) cortex, whereas the axons of NNS and tactile neurons projected chiefly to the insular cortex and the amygdala.In conclusion, our results demonstrate a spinal-PoT-S2/insular cortices nociceptive pathway that conveys nociceptive messages arising from lamina I and spinal neurons of deep laminas. Furthermore, our results demonstrate for the first time that projections of PoT neurons are correlated to their physiological properties.

From 1968 to 1993, 179 newly diagnosed patients with polycythemia vera (PV) were enrolled in a prospective study using pipobroman as first chemotherapy. Among them, 140 fulfilled the Polycythemia Vera Study Group criteria for PV, and 39 patients (22%) can be considered as idiopathic erythrocytosis (IE). Vascular events occurred in 10% of IE and 20% of PV patients and solid tumors in 7.7% of IE and 12.8% of PV patients. There were no differences between PV and IE patients with regard to progression to myelofibrosis (MF), leukemic events and overall survival. Overall, 98.3% of patients initially responded to pipobroman, with very mild toxicity. A total of 164 PV patients who received more than 1 year of pipobroman were analyzed for long-term evolution. The actuarial risk of thrombosis was 15.6 and 23.8% at 10 and 18 years, respectively. In all, 21 patients developed a solid tumor during follow-up, added and/or switched drugs being a risk factor. Actuarial risk of MF was as low as 4.9 and 9.4% at 10 and 15 years, respectively. Actuarial risk of leukemia was 14.4 and 18.7% at 10 and 15 years, respectively. Hyperleukocytosis at diagnosis was the only variable significantly associated with higher risk of leukemia. The median survival was 15.5 years, with two initial adverse prognostic factors: age above 60 years and hyperleukocytosis. Despite an increasing risk of leukemia with time, survival was not lower when compared to the French matched population. Only age and hyperleukocytosis at diagnosis were found to have a prognostic value in PV.

Projections to the forebrain from lamina I of spinal and trigeminal dorsal horn were labeled anterogradely with Phaseolus vulgaris-leucoagglutinin (PHA-L) and/or tetramethylrhodamine-dextran (RHO-D) injected microiontophoretically. Injections restricted to superficial laminae (I/II) of dorsal horn were used primarily. For comparison, injections were also made in deep cervical laminae. Spinal and trigeminal lamina I neurons project extensively to restricted portions of the ventral posterolateral and posteromedial (VPL/VPM), and the posterior group (Po) thalamic nuclei. Lamina I also projects to the triangular posterior (PoT) and the ventral posterior parvicellular (VPPC) thalamic nuclei but only very slightly to the extrathalamic forebrain. Furthermore, the lateral spinal (LS) nucleus, and to a lesser extent lamina I, project to the mediodorsal thalamic nucleus. In contrast to lamina I, deep spinal laminae project primarily to the central lateral thalamic nucleus (CL) and only weakly to the remaining thalamus, except for a medium projection to the PoT. Furthermore, the deep laminae project substantially to the globus pallidus and the substantia innominata and more weakly to the amygdala and the hypothalamus. Double-labeling experiments reveal that spinal and trigeminal lamina I project densely to distinct and restricted portions of VPL/VPM, Po, and VPPC thalamic nuclei, whereas projections to the PoT appeared to be convergent. In conclusion, these experiments indicate very different patterns of projection for lamina I versus deep laminae (III-X). Lamina I projects strongly onto relay thalamic nuclei and thus would have a primary role in sensory discriminative aspects of pain. The deep laminae project densely to the CL and more diffusely to other forebrain targets, suggesting roles in motor and alertness components of pain.

The lateral part of the ventromedial thalamus (VM l) relays nociceptive inputs from the whole body surface to the dorsolateral frontal cortex. The aim of the present study was to investigate the effects of systemic morphine on nociceptive activity evoked in VM l neurones either by thermal (48 degrees C) or by supramaximal percutaneous electrical stimuli. The noxious thermal evoked responses were depressed by 10.8 +/- 10.1%, 48.3 +/- 23.0% and 67.3 +/- 10.1%, 5 min after i.v. injections of 1.0, 1.73 and 3.0 mg/kg of morphine, respectively. Moreover, strong depressive effects on the Adelta- and C-fibre responses were already present 5 min after the injection. The responses were significantly reduced by 7.2 +/- 5.9%, 32.5 +/ 11.1% and 37.2 +/- 11.8% for Adelta fibres after i.v. injections of 1.0, 1.73 and 3.0 mg/kg of morphine, respectively. The corresponding values for C-fibre evoked responses were 16.3 +/- 16.2%, 57.0 +/- 12.0% and 69.0 +/- 8.2%. The dose of morphine that reduced VM l neuronal nociceptive responses by 50% (1.73 mg/kg) was around 3.5 times lower than that necessary to inhibit the responses of its spinal or medullary relays under similar experimental conditions. These results, added to the data of the literature, suggest that supraspinal effects of morphine are primarily mediated at the thalamic level. It is tempting to speculate that morphine-induced reductions of attentional or psychomotor responses related to pain may be mediated by its action on VM l.

Serotoninergic innervation of the central nervous system is provided by hindbrain raphe nuclei (B1–B9). The extent to which each raphe subdivision has distinct topographic organization of their projections is still unclear. We provide a comprehensive description of the main targets of the rostral serotonin (5-HT) raphe subgroups (B5–B9) in the mouse brain. Adeno-associated viruses that conditionally express GFP under the control of the 5-HT transporter promoter were used to label small groups of 5-HT neurons in the dorsal (B7d), ventral (B7v), lateral (B7l), and caudal (B6) subcomponents of the dorsal raphe (DR) nucleus as well as in the rostral and caudal parts of the median raphe (MR) nucleus (B8 and B5, respectively), and in the supralemniscal (B9) cell group. We illustrate the distinctive and largely non-overlapping projection areas of these cell groups: for instance, DR (B7) projects to basal parts of the forebrain, such as the amygdala, whereas MR (B8) is the main 5-HT source to the hippocampus, septum, and mesopontine tegmental nuclei. Distinct subsets of B7 have preferential brain targets: B7v is the main source of 5-HT for the cortex and amygdala while B7d innervates the hypothalamus. We reveal for the first time the target areas of the B9 cell group, demonstrating projections to the caudate, prefrontal cortex, substantia nigra, locus coeruleus and to the raphe cell groups. The broad topographic organization of the different raphe subnuclei is likely to underlie the different functional roles in which 5-HT has been implicated in the brain. The present mapping study could serve as the basis for genetically driven specific targeting of the different subcomponents of the mouse raphe system. Electronic supplementary material The online version of this article (doi:10.1007/s00429-014-0924-4) contains supplementary material, which is available to authorized users.

International audience; A new architectural study of the stadium of Domitian, as well as archaeological research work in and around Piazza Navona, leads to reconsider some of the hypothesis presented in 1943 by A. M. Colini. This new analysis takes advantage of recent studies on Stadium architecture all over the Empire and of an improved knowledge of the topography of the Campus Martius. The Stadium of Domitian was conceived as a deeply original building. An elitist monument, it is one of the most accomplished models in the category of spectacles building. The way it was conceived, and especially the organisation of the substructure supporting the seats, made easier the re-use of a building, which has always played a major role in the urban organisation of central Rome. Modern Piazza Navona, which keeps the formal memory of the ancient stadium, is towered by the cupola of Boromini’s church of S. Agnese, at the centre of the western long side. The authors show how its specific architectural features predisposed this location to host the earliest shrine dedicated to the young martyr.; La reprise du dossier architectural du stade de Domitien, ainsi que l’apport de nouvelles recherches menées dans les caves de l'immeuble occupé par l'EfR (62, place Navone) et sur le pourtour de la place ont permis de renouveler certaines hypothèses formulées en 1943 par A. M. Colini. Cette nouvelle analyse profite également de récentes études sur l'architecture des stades et d'une meilleure connaissance de la topographie du Champ de Mars. Le stade de Domitien révèle toute l'originalité de sa conception architecturale. Édifice élitiste, il constitue l'un des exemples les plus aboutis des monuments de spectacle. Sa conception d'ensemble et tout particulièrement l'organisation de la structure qui supportait ses gradins ont favorisé la réutilisation d'un monument qui n'a cessé de jouer un rôle important dans l'urbanisme du cœur de Rome. L'actuelle piazza Navona, qui conserve la mémoire de l'édifice antique, est dominée par la coupole de l'église Sainte- Agnès située au centre du long côté Ouest. Les auteurs montrent comment certaines caractéristiques architecturales du stade de Domitien ont prédisposé cet emplacement particulier à accueillir le premier lieu de culte dédié à la jeune martyre.

The lateral capsular division (CeLC) of the central nucleus (Ce) of the amygdala, in the rat, has been shown to be the main terminal area of a spino(trigemino)-parabrachio-amygdaloid nociceptive pathway [Bernard & Besson (1990) J. Neurophysiol. 63, 473-490; Bernard et al. (1992) J. Neurophysiol. 68, 551-569; Bernard et al. (1993) J. Comp. Neurol. 329, 201-229]. The projections to the forebrain from the CeLC and adjacent regions were studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L) restricted in subdivisions of the Ce and the basolateral amygdaloid nucleus anterior (BLA). Our data showed that the entire CeLC projects primarily and extensively to the substantia innominata dorsalis (SId). The terminal labelling is especially dense in the caudal aspect of the SId. The other projections of the CeLC in the forebrain were dramatically less dense. They terminate in the bed nucleus of the stria terminalis (BST) and the posterior hypothalamus (pLH). No (or only scarce) other projections were found in the remaining forebrain areas. The Ce lateral division (CeL) and the Ce medial division (CeM), adjacent to the CeLC, also project to the SId with slightly lower density labelling. However, contrary to the case of the CeLC, both the CeL and the CeM extensively project to the ventrolateral subnucleus of the BST (BSTvl) with a few additional terminals found in other regions of the lateral BST. Only the CeM projects densely to both the interstitial nucleus of the posterior limb of the anterior commissure and the caudal most portion of the pLH. The projections of the BLA are totally different from those of the Ce as they terminate in the dorsal striatum, the accumbens nucleus, the olfactory tubercle, the nucleus of olfactory tract and the rostral pole of the cingulate/frontal cortex. This study demonstrates that the major output of the nociceptive spino(trigemino)-parabrachio-CeLC pathway is to the SId. It is suggested that the CeLC-SId pathway could have an important role in anxiety, aversion and genesis of fear in response to noxious stimuli.

The somatosensory properties of ventromedial (VM) thalamic neurons were investigated in anesthetized rats by examining their responses to calibrated cutaneous stimuli. A population of neurons within the lateral part of the ventromedial thalamus (VMl) showed two peaks of activation after percutaneous electrical stimuli, regardless of which part of the body was stimulated. The early and late peaks were elicited by Aδ- and C-fiber activities with mean conduction velocities of 12.9 ± 0.9 and 1 ± 0.2 m/sec, respectively. These responses were strongly depressed or blocked after microinjections within the medullary subnucleus reticularis dorsalis of xylocaine or the NMDA antagonist MK-801. None of the VMlneurons responded to innocuous cutaneous or proprioceptive stimuli. In contrast, all these neurons responded to noxious mechanical and thermal stimulation of the limbs and showed monotonic increases in their discharges to increasingly strong noxious cutaneous stimuli. In addition, some VMlneurons were antidromically activated by stimulation in layer I of the dorsolateral frontal cortex. These findings suggest that the rat VMlconveys and encodes cutaneous nociceptive inputs from any part of the body surface to layer I of the dorsolateral neocortex. This reticulo-thalamo-cortical network may allow any signal of pain to gain access to widespread areas of the neocortex and thus help prime the cortex for attentional reactions and/or the coordination of motor responses.

The organization of projections from the parabrachial (PB) area to the ventral posterior parvicellular (VPpc) "gustatory" and intralaminar nuclei of the thalamus was studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L), into subregions of the PB area. The present study is a follow-up of three former studies (Bernard et al. [1993] J. Comp. Neurol. 329:201-229; Alden et al. [1994] J. Comp. Neurol. 341:289-314; Bester et al. [1997a] J. Comp. Neurol. 383:245-281) that examined PB projections onto the amygdala, the bed nucleus of the stria terminalis, and the hypothalamus. Our data showed that (1) the region centered in the internal lateral PB subnucleus projects densely with a bilateral and symmetric pattern to the caudal portion of the paracentral and, to a lesser extent, to the adjacent portion of the central and parafascicular medial thalamic nuclei; (2) the mesencephalic PB region centered in the ventral lateral subnucleus and scattered neurons in the subjacent brachium conjunctivum project primarily, although diffusely, to the central medial thalamic nucleus. The third region includes two subgroups: (3a) the medial subgroup, including the medial, the waist area, and the ventral lateral subnuclei of the pontine PB area, projects bilaterally but with a weak ipsilateral predominance to the VPpc, terminals bearing large varicosities. Additionally, a diffuse projection with small varicosities spreads in the area between the two VPpc nuclei and the central medial nucleus. (3b) The lateral subgroup, centered in the external medial subnucleus, projects with a contralateral predominance in the periphery of the VPpc nuclei, most terminals being located around the dorsomedial tip. It is suggested that the PB projections to the intralaminar nucleus could be involved in cortical limbic arousal processing in relation with nociceptive, (somatic, visceral, and intraoral) and gustatory aversive stimuli. The projection with large varicosities inside the VPpc could process gustatory discrimination.

The effect of graded inflammatory stimuli (intraplantar-carrageenan, 0.2, 1, and 6 mg/150 microl) on paw edema and c-Fos protein expression at two levels of the spinoparabrachial pathway, the spinal cord and parabrachial area (PB), were studied. The present study, in awake rats, is an extension of previous study (Bester et al. [1997] J. Comp. Neurol. 383:439-458) which evaluated, in anesthetized rats, the effect of graded cutaneous heat stimulation on c-Fos-expression at the same levels. At the spinal level, the c-Fos-protein-like-immunoreactive (c-Fos-LI) neurons were located primarily in superficial laminae ipsilateral to intraplantar carrageenan. The number of c-Fos-LI neurons increased dose dependently (r = 0.973, n = 24) for carrageenan, from a number close to zero for the saline injection. At the PB level, c-Fos was predominantly expressed contralateral to intraplantar carrageenan. c-Fos-LI neurons were located primarily around the pontomesencephalic junction in (i) a restricted pontine area, centered in the lateral crescent, and including an adjacent part of the outer portion of the external lateral subnucleus, and (ii) the mesencephalic superior lateral subnuclei. The number of c-Fos-LI neurons in the PB area was correlated with that in the superficial laminae (r = 0.935, n = 24) and with the paw edema (r = 0.931, n = 24). No significant changes in c-Fos expression were observed in the nucleus of the solitary tract and ventrolateral medulla. The close correlation between c-Fos expression at both the spinal and PB levels and inflammatory edema provides further evidence for the involvement of spinoparabrachial pathway in inflammatory nociceptive processes. The present results are congruent with the existence of electrophysiologically demonstrated spinoparabrachio-amygdaloid and -hypothalamic nociceptive pathways.

The distribution and organization of diencephalic projections from the subnucleus reticularis dorsalis (SRD) and the neighbouring cuneate nucleus (Cu) were studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin in SRD and Cu and wheat germ agglutinin-apo horseradish peroxidase-gold in some selected thalamic areas. As previously reported, the efferent projections from the Cu were essentially contralateral and terminated mainly in the ventroposterolateral thalamic nucleus. Less dense terminals from the Cu were also observed in the posterior thalamic group, the ventral aspect of the zona incerta and the caudal and dorsal portion of the reuniens area. Retrograde tracer injections in the medial ventroposterolateral thalamic nucleus labeled numerous cells in the contralateral Cu, with a smaller number in the gracile nucleus. From the SRD, terminals were observed in the lateral aspect of the ventromedial thalamic nucleus, the lateral parafascicular area and, to a lesser extent, in the ventral aspect of the zona incerta and the core of the reuniens area. Retrograde tracer injections in the lateral part of the ventromedial thalamic nucleus labeled cells in the caudal medulla, many of which were located in the dorsal-most aspect of the SRD throughout its caudo-rostral extent. The existence of SRD-thalamic connections reinforces the idea that the caudal reticular formation is an important nociceptive relay to the thalamus. Our data shed new light on old hypotheses suggesting that, in addition to spino-thalamic pathways, spino-reticulo-thalamic pathways may play an important role in distributing pain signals to the forebrain.

We have analysed in briefly anaesthetised rats (1% halothane for 18 minutes) the effects of innocuous and noxious heat, applied to the hindpaw, on evoked c-Fos immunoreactivity at the levels of the parabrachial area (PB), spinal cord, and nucleus of the solitary tract (NTS). After anaesthesia recovery, animals were left to move freely for 2 hours. At the spinal level, c-Fos was expressed primarily in the ipsilateral superficial laminae, increasing with the applied temperatures in a dependent manner in the noxious range (correlation coefficient r = 0.954, n = 20). At the NTS level, no noxiously evoked c-Fos expression was observed. At the PB level, c-Fos was expressed preferentially contralaterally, increasing with the applied temperatures in a dependent manner in the noxious range (r = 0.971, n = 25). The maximum expression was observed in the outer portion of the external lateral, the lateral crescent, and the superior lateral subnuclei around the pontomesencephalic junction. This was congruent with the densest supraspinal projection of lamina I neurones of the dorsal horn. Labelling in the PB area was highly correlated (r = 0.936, n = 20) with labelling in the superficial laminae. We conclude that, under our experimental procedures, noxious heat-induced c-Fos expression at the PB level depends on the intensity of the noxious stimulation. These data further support the relevance of the recently described spino-PB pain pathway. Because of their location, the Fos-immunoreactive neurones observed in the pontine and the mesencephalic divisions of the PB area were likely PB-amygdaloid and PB-hypothalamic nociceptive neurones, respectively.

The organization of projections from the parabrachial (PB) area to the hypothalamus was studied in the rat by using microinjections of Phaseolus vulgaris-leucoagglutinin (PHA-L) into subregions of the PB area. The present study is a follow-up of two former studies (Bernard et al. [1993] J. Comp. Neurol. 329:201-229; Alden et al. [1994] J. Comp. Neurol. 341:289-314) that examined PB projections onto the amygdala and the bed nucleus of the stria terminalis. The results demonstrate that 1) the mesencephalic PB region, centered in the lateral portion of the superior lateral subnucleus projects extremely densely to almost the entire dorsomedial subdivision of the ipsilateral ventromedial hypothalamic nucleus; 2) the mesencephalic PB region, located in the medial portion of the superior lateral subnucleus and weakly overflowing into the rostralmost dorsal lateral pontine subnucleus, projects densely to the retrochiasmatic area and, to a lesser extent, to the ipsilateral ventromedial nucleus of the hypothalamus; 3) the PB region, including the central lateral, a portion of the superior lateral, and the outer external lateral subnuclei, projects densely to the ipsilateral median, anteroventral, and periventricular preoptic hypothalamic nuclei and projects more weakly to the dorsal border of the paraventricular nucleus (PVN). No consistent projection was found in the magnocellular PVN. All of these PB regions also project diffusely to the dorsomedial area and to a small tuberal subfornical hypothalamic area. In addition, the medial half of the PB area projects consistently to the posterior lateral hypothalamus. It is suggested that these pathways may be involved in aversive-defensive behavior, in autonomic and neuroendocrine aspects of pain, and in feeding and energy metabolism regulation. J. Comp. Neurol. 383:245-281, 1997. © 1997 Wiley-Liss, Inc.

1. Thirty-five percent of 120 neurons recorded extracellularly in the parabrachial (PB) area of anesthetized rats responded to a peripheral cold stimulus (0 degrees C). The cold-sensitive neurons were located in the lateral PB area, and most of those exhibiting a strong response to cold stimuli were inside or in close vicinity to the area receiving a high density of projections from superficial neurons of the dorsal horn. 2. The receptive fields for cold stimulation often were restricted to one or two parts of the body with a contralateral predominance for the limbs. No side predominance was observed for the face. 3. From a low spontaneous activity (10th percentile < median < 90th percentile: 0.1 < 1.5 < 5 Hz), the PB neurons responded to cold noxious stimuli (0 degree C water bath or waterjet, 20 s), without observable delay, with a sustained discharge. The mean maximal response to the stimulus was 16.1 +/- 1.2 Hz (mean +/- SE; n = 42). 4. About one-half (45%) of these cold-sensitive neurons were activated specifically by cold stimulation and did not respond or were inhibited by noxious heat and/or pinch. The remaining (55%) cold-sensitive neurons were also driven by heat and/or pinch. 5. The cold-sensitive neurons exhibited a clear capacity to encode cold stimuli in the noxious range: the stimulus-response function was always positive and monotonic from 30 to 0 degrees C; the mean curve was linear between 20 and 0 degrees C before plateauing between 0 to -10 degrees C; the mean threshold to cold stimulation was 17.1 +/- 1 degrees C (n = 21) and the mean t50 was 10.7 +/- 1.1 degrees C (n = 13). 6. The cold-sensitive neurons responded to intense transcutaneous electrical stimulation with an early and/or a late peak of activation, the latencies of which were in the 15-50 ms and 80-170 ms ranges (n = 8), respectively, i.e., compatible with the activation of A delta and C fibers. Interestingly, the cold-specific neurons predominantly responded with a late peak, suggesting these neurons were primarily driven by peripheral C fibers. 7. The intravenous injection of morphine depressed the responses of PB neurons to cold noxious stimuli in a dose-related (1, 3, and 9 mg/kg) and naloxone reversible fashion. The ED50 value was estimated approximately 2 mg/kg. Furthermore, two populations of neurons could be separated according to their morphine sensitivity. 8. It is concluded that PB cold-nonspecific neurons could be involved in affective-emotional, autonomic and neuroendocrine reactions in response to noxious cold events. The PB cold-specific neurons could be, in addition, involved in some thermoregulatory processes.

1. Parabrachiohypothalamic (PB-H) neurons (n = 71) were recorded with extracellular micropipettes in the parabrachial (PB) area and were antidromically driven from the ventromedial nucleus (VMH) or the retrochiasmatic area (RCh) of the hypothalamus, in the anesthetized rat. The spontaneous activity of these neurons was very low, (10th percentile < median frequency < 90th percentile were 0.01 < 0.2 < 7 Hz). The axons of these neurons exhibited a very slow conduction velocity in the range of 0.2-1.4 m/s, i.e., corresponding to thin unmyelinated fibers. 2. Most PB-H neurons (89%) were located in the mesencephalic division of the PB area (mPB) mainly in the superior lateral (mPBsl) and external lateral (mPBel) subnuclei. 3. These units were separated in three groups: 1) a group of nociceptive-specific (NS) neurons (49%) activated by mechanical and/or thermal (heat) cutaneous stimuli only in noxious range; 2) a group of inhibited neurons (7%), not activated by any of the mechanical or thermal cutaneous stimuli but inhibited, by at least one of these stimuli, which had to be in noxious range; and 3) a group of nonresponsive neurons (44%). 4. The NS neurons responded exclusively to mechanical (pinch or squeeze) and/or thermal (waterbath or waterjet > 44 degrees C) noxious stimuli with a rapid onset, a marked and sustained activation, and generally no afterdischarge. The magnitude of the responses was between 2 and 30 Hz with a mean value of 14.8 +/- 1.4 Hz (mean +/- SE, n = 49). These neurons exhibited a clear capacity to encode thermal stimuli in the noxious range: 1) the stimulus-response function was always positive and monotonic; 2) the slope of the mean curve increased up to a maximum (between 46 and 50 degrees C) then beyond the slope decreased; and 3) the mean threshold was 44.3 +/- 2.2 degrees C. 5. The excitatory receptive fields of the NS neurons were often large including all (22% of the population) or several (67% of the population) parts of the body. In the few remaining cases (11%) they were restricted to one part of the body. In addition, in several cases, noxious stimuli applied outside the excitatory receptive field were found to strongly inhibit the discharge of NS neurons. 6. Most NS neurons responded to intense transcutaneous electrical stimulation with two peaks of activation.(ABSTRACT TRUNCATED AT 400 WORDS)

The organization of efferent projections from the pontine parabrachial (pPB) area to the forebrain rostral to the central nucleus of the amygdala (Ce) was studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L), into subregions of the pPB area. The present study is a follow-up of a former study (Bernard et al. [1993] J. Comp. Neurol. 329:201-229) which examines pPB projections onto the Ce. The results demonstrate that: (1) the pPB(m) region (the medial, the ventral lateral subnuclei and the waist area) diffusely projects to the lateral division (BSTL) of the bed nucleus of the stria terminalis (BST), the Ce-BSTL continuum (including, the dorsal portion of substantia innominata, the ventral portion of globus pallidus, the fundus striatum, and the substriatal area) and to a lesser extent the agranular insular cortex; (2) the pPB(1) region [the central lateral (pPBcl) and the outer portion of external lateral subnuclei] densely projects to the dorsal lateral subnucleus of BST (BSTdl); only the pPBcl subnucleus projects to the median, the anteroventral and the periventricular nuclei of the preoptic hypothalamus; and (3) the remaining pPB area (the dorsal lateral, part of the external lateral and the external medial subnuclei) projects to the nucleus of horizontal limb of diagonal band but does not project onto the BST and the preoptic hypothalamus. It is suggested that the pPB(m)-BSTL "diffuse pathway" is mainly implicated in motivational and autonomic aspects of taste. The pPB(1)-BSTdl and hypothalamic "concentrated pathways" could be implicated in autonomic and nociceptive processes.

It has recently been demonstrated that the nucleus centralis of the amygdala contains numerous neurons specifically driven by noxious stimuli. The aim of the present study was to investigate the effect of i.v. morphine on responses of neurons located in the nucleus centralis of the amygdala to noxious mechanical or thermal stimuli. It was observed, in halothane-anesthetized rats, that i.v. morphine caused a marked depression of responses induced by noxious thermal (waterbath, 50 degrees C) and mechanical (pinch) stimuli and caused a moderate depression of spontaneous activity in a dose-related (1, 3, 9 mg/kg) and naloxone reversible fashion. The ED50 value was 1.2 and 9 mg/kg for i.v. morphine for the evoked activity and spontaneous activity, respectively. The strong depressive effect of morphine on evoked activity probably reflects a direct action of this drug at both spinal and parabrachial levels. These results could account, at least in part, for the effect of morphine on the emotional-affective aspects of pain.

The 5-HT1A receptor (5-HT1AR) plays a key role in the inhibitory influence of serotonin (5-HT) on rapid eye movement (REM) sleep in rodents. However, the neuronal networks mediating such influence are mostly unknown, notably in the mouse. This led us to map 5-HT1AR mRNA, by in situ hybridization histochemistry (ISHH), and to characterize the neuronal phenotype of 5-HT1AR mRNA-positive neurons by dual ISHH and ISHH combined with immunohistochemistry, throughout the mouse rostral brainstem, a pivotal region for the generation of REM sleep and cortical activation. 5-HT1AR mRNA was found in most 5-HT neurons in the dorsal raphe (DR), the median raphe (MnR), the B9, and the interpeduncular (IP) nuclei. 5-HT1AR mRNA-positive neurons were also identified in individualized clusters of γ-aminobutyric acid (GABA)ergic neurons in the DR and in neurons of an undetermined phenotype in the MnR. In addition, 1) GABAergic neurons of the ventral portion of Gudden's dorsal tegmental nucleus (DTg), the IP, and the caudal portion of the deep mesencephalic nucleus (DpMe), and 2) glutamatergic neurons scattered in the caudal pontine reticular nucleus (PnC) and densely packed in the internal lateral parabrachial subnucleus (PBil) also expressed 5-HT1AR mRNA. In contrast, no specific 5-HT1AR-related ISHH signal was generally detected in brainstem cholinergic and catecholaminergic neurons. These results emphasize the role of 5-HT1AR as an autoreceptor and the phenotypical heterogeneity of 5-HT1AR-expressing neurons within the DR and the MnR in the mouse brain. They also provide a neuroanatomical basis for understanding the influence of 5-HT1AR on REM sleep and wakefulness. J. Comp. Neurol. 518:2744–2770, 2010. © 2010 Wiley-Liss, Inc.

1. Neurons (n = 177) were recorded with extracellular micropipettes in and around the nucleus centralis of the amygdala (Ce), in anesthetized rats. The spontaneous activity of these neurons was variable (0.25 less than 3 less than 35 Hz, n = 175; 10th percentile less than median less than 90th percentile). A majority (80%) of these neurons were excited or inhibited exclusively or preferentially by noxious stimuli. These units were separated into two groups: 1) a group of neurons excited by noxious stimuli (46% of the whole population) and 2) a group of neurons inhibited by noxious stimuli (34% of the whole population). 2. The receptive fields of both groups of neurons were very large: in about one-half the cases the neurons responded similarly from all parts of the body, and in the other cases the responses were greater when the stimuli were applied to a restricted part of the body. 3. Seventy-seven percent of the excited neurons had responses of relatively high magnitudes. In this group, most cells (75%) were exclusively driven by noxious stimuli; the others (25%) were preferentially activated by noxious stimuli. These neurons responded to mechanical (pinch or squeeze) and/or thermal (water bath or water jet greater than 44 degrees C) noxious stimuli with a marked and sustained activation. 4. Sixty percent of the inhibited neurons had a marked decrease of activity in response to noxious stimuli. In this group, most of them (81%) were exclusively inhibited by noxious stimuli, whereas the remainder (19%) were preferentially inhibited by noxious stimuli. These neurons responded to mechanical (pinch or squeeze) and/or thermal (water bath or waterjet greater than 44 degrees C) noxious stimuli with a suppression or a marked and sustained decrease in activity. 5. All of the nociceptive neurons responded to intense transcutaneous electrical stimulation with one or several components of activation or inhibition. According to their latencies, three types of components were distinguished: early, intermediate, and late components. We estimate that the early and the intermediate components would be triggered by the activity of peripheral fibers in the 6- to 20-m/s range and therefore could be in the A delta fibers range, whereas the late component would be triggered by fibers in the 0.5- to 1-m/s range and therefore could be in the C fibers range. 6. The neurons excited or inhibited by noxious stimuli were not homogeneously distributed in and around the Ce.(ABSTRACT TRUNCATED AT 400 WORDS)

Stimulation of the dorsolateral periaqueductal gray matter (dlPAG) and the B3 cell group inhibits the cardiovagal component of the baroreflex in rats. Our aim was to determine whether the defence reaction induces similar modulatory effects on the cardiac response of the von Bezold-Jarisch reflex and the carotid chemoreceptor reflex. We examined the effects of dlPAG stimulation on the reflex bradycardia triggered by systemic administration of phenylbiguanide or potassium cyanide. Electrical and chemical stimulation of the dlPAG produced marked inhibition of the cardiovagal components of the von Bezold-Jarisch and the carotid chemoreceptor reflexes. In addition, as 5-HT(3), NK(1) and GABA(A) receptor activation blocks cardiac reflex responses, we studied whether these receptors were involved in the dlPAG-induced inhibitory effects. We found that, after microinjection of granisetron (a 5-HT(3) receptor antagonist), bicuculline (a GABA(A) receptor antagonist) and GR-205171 (an NK(1) receptor antagonist) into the nucleus of the solitary tract (NTS), reflex bradycardic responses were preserved during dlPAG stimulation. Finally, activation of the B3 region also inhibited both reflex bradycardic responses, and these effects were prevented by prior blockade of 5-HT(3) receptors in the NTS. The inhibitory effect of dlPAG stimulation on the cardiac reflex responses was prevented by inhibition of neurons in the medullary B3 region. In conclusion, 5-HT(3), GABA(A) and NK(1) receptors in the NTS appear to be involved in the inhibition of the von Bezold-Jarisch reflex and the carotid chemoreceptor reflex bradycardia evoked by activation of neurons in the dlPAG and the raphe magnus.

The present study was designed to identify the neuronal mechanisms causing cardiac baroreflex inhibition associated with thermal nociception in rats. Under urethane-anesthesia, noxious thermal stimulior = 48 degrees C were found to inhibit the cardiac baroreflex, whereas noxious stimulior = 46 degrees C had no effect. Using double immunohistochemical labeling, noxious stimulior = 48 degrees C were found to evoke primarily a strong expression of Fos protein (Fos) encoded by c-fos gene in serotonergic neurons of lateral paragigantocellular reticular nucleus (LPGi). Noxious stimulior = 46 degrees C did not evoke Fos expression in any serotonergic neurons of the brainstem. Local blockade of neuronal activity by bilateral microinjections of fluorescent muscimol (a GABA(A) receptor agonist tagged with a fluorophore that allowed visualization of the injections) into both the LPGi and the raphe magnus nucleus prevented the inhibitory effect of noxious stimulior = 48 degrees C on the cardiac baroreflex. Bilateral microinjections of granisetron (a 5-HT(3) antagonist) within the nucleus tractus solitarius also prevented the inhibition of cardiac baroreflex elicited by noxious stimulior = 48 degrees C. These results show that activation of serotonergic cells in the LPGi is critical to trigger nucleus tractus solitarius-mediated cardiac baroreflex inhibition elicited by intense thermal noxious stimuli.

1. Neurons were recorded in the parabrachial (PB) area, located in the dorsolateral region of the pons (with the use of extracellular micropipette), in the anesthetized rat. Parabrachioamygdaloid (PA) neurons (n = 67) were antidromically identified after stimulation in the centralis nucleus of the amygdala (Ce). The axons of these neurons exhibit a very slow conduction velocity, between 0.26 and 1.1 m/s, i.e., in the unmyelinated range. 2. These PA neurons were located in a restricted region of the PB area: the subnuclei external lateral (PBel) and external medial (PBem). A relative somatotopic organization was found in this region. 3. These units were separated into two groups: 1) a group of nociceptive-specific (NS) neurons (69%), which responded exclusively to noxious stimuli, and 2) a group of nonresponsive (NR) neurons (31%). 4. The NS neurons exhibited low or lacked spontaneous activity. They responded exclusively to mechanical (pinch or squeeze) and/or thermal (waterbath or waterjet greater than 44 degrees C) noxious stimuli with a marked and sustained activation with a rapid onset and generally without afterdischarge. Noxious thermal stimuli generally induced a stronger response than the noxious mechanical stimuli. These neurons exhibited a clear capacity to encode thermal stimuli in the noxious range: 1) the stimulus-response function was always positive and monotonic; 2) the slope of the curve progressively increased up to a maximum where it was very steep, then the steepness of the slope decreased close to the maximum response; and 3) the mean threshold was 44.1 +/- 2 degrees C, and the point of steepest slope of the mean curve was around 47 degrees C. 5. The excitatory receptive fields of the NS neurons were large in the majority (70%) of the cases and included several areas of the body. A more marked activation was often obtained from stimuli applied to one part of the body, denoted as the preferential receptive field (PRF). In the other cases (30%), the excitatory receptive field was relatively small (SRF) and restricted to one part of the body (the tail, a paw, a hemiface, or the tongue). Both the PRF and SRF were more often located on the contralateral side. In addition, noxious stimuli applied outside the excitatory receptive field were found to strongly inhibit the responses of NS neurons. 6. All the NS neurons responded to intense transcutaneous electrical stimulation applied to the PRF or SRF with two peaks of activation.(ABSTRACT TRUNCATED AT 400 WORDS)

The present study was designed to identify the serotonergic pathway causing baroreflex inhibition associated with the defense reaction in rats. Under conditions that produce physiological responses typical of the defense reaction, electrical stimulation of the dorsal periaqueductal gray (dPAG) was found to double c-Fos immunoreactive serotonergic neurons within the midrostrocaudal extent of the B3 group (which comprises the raphe magnus and the lateral paragigantocellular reticular nuclei) in anesthetized rats. Local blockade of neuronal activity by microinjection of muscimol (a GABAA receptor agonist) directly into the B3 region prevented the inhibitory effect of dPAG activation on the cardiac baroreflex. Conversely, neuron activation by local application of D,L-homocysteic acid into B3 region caused baroreflex inhibition that was suppressed by microinjection of granisetron (a 5-HT3 antagonist) into the nucleus tractus solitarius. These results show that activation of serotonergic cells in the mid-portion of B3 group is critical to trigger baroreflex inhibition occurring during the defense reaction evoked by dPAG stimulation. J. Comp. Neurol. 506:108 –121, 2008. © 2007 Wiley-Liss, Inc. Indexing terms: heart rate; serotonin; c-Fos; tractus solitarius; periaqueductal; ventromedial medulla

Objectif La regulation des etats de vigilance par la serotonine (5-HT) s’effectue notamment au travers des recepteurs 5-HT1A. Ainsi, les agonistes de ces recepteurs induisent un eveil marque, principalement au detriment du sommeil paradoxal. Nous avons prealablement identifie une population de neurones du tronc cerebral qui exprime fortement ces recepteurs : les neurones GABAergiques du tegmentum dorsal de Gudden (DTg). Nous proposons que ce noyau participe a l’action de la 5-HT sur les etats de vigilance. Methodes Afin de tester cette hypothese, nous avons evalue les consequences de l’inactivation du DTg sur les etats de vigilance chez la souris suite a : (1) l’administration locale d’un agoniste specifique des recepteurs 5-HT1A (le 8-OH-DPAT) et (2) des lesions electrolytiques. Afin d’identifier les structures cibles du DTg, differents tracages anatomiques ont ete realises chez la souris. Resultats Chez la souris, la micro-injection locale de 8-OH-DPAT dans le DTg facilite la survenue de l’eveil au detriment du sommeil paradoxal. La specificite d’action du 8-OH-DPAT vis-a-vis des recepteurs 5-HT1A a ete validee chez des souris chez lesquelles l’effet local du 8-OH-DPAT a ete bloque par l’injection systemique prealable d’un antagoniste specifique des recepteurs 5-HT1A, le WAY 100635. La lesion bilaterale du DTg augmente l’eveil pendant la phase lumineuse sans affecter le sommeil paradoxal. La mise en œuvre de techniques de tracage anterograde nous a permis de montrer que les neurones du DTg projettent exclusivement au niveau de l’hypothalamus mamillaire lateral. Nous avons ensuite confirme ces donnees par tracage retrograde chez des souris transgeniques dont l’expression du recepteur 5-HT1A peut etre visualisee par une simple coloration (X-gal). Conclusion Ces donnees mettent en avant un role nouveau du DTg dans la modulation des etats de vigilance par la 5-HT. Elles permettent egalement de proposer que le circuit « noyaux du raphe/DTg/hypothalamus mamillaire » participe a la regulation des etats de vigilance. Dans ce cadre, les neurones GABAergiques du DTg faciliteraient la survenue du sommeil en exercant une pression negative sur les neurones promoteurs d’eveil de l’hypothalamus posterieur.