Temporomandibular disorders (TMD) are a family of prevalent chronic pain disorders affecting masticatory muscles and/or the temporomandibular joint. There is no unequivocally recognized peripheral aetiology for idiopathic TMD. The central nervous system (CNS) may initiate and/or maintain the pain in idiopathic TMD due to sustained or long-term nociceptive input that induces maladaptive brain plasticity, and/or to inherent personality-related factors that may reduce the brain's capacity to modulate nociceptive activity. The main aim of this thesis is to determine whether there are structural neural abnormalities in patients with TMD, and whether these abnormalities are related to TMD pain characteristics, or to neuroticism. The specific aims are to delineate in TMD: (1) gray matter (GM) brain abnormalities and the contribution of pain and neuroticism to abnormalities; (2) the contribution of abnormal brain GM aging in focal cortical regions associated with nociceptive processes; and (3) abnormalities in brain white matter and trigeminal nerve and the contribution of pain. In groups of 17 female patients with TMD and 17 age- and sex- matched controls, magnetic resonance imaging revealed that patients with TMD had: (1) thicker cortex in the somatosensory, ventrolateral prefrontal and frontal polar cortices than controls, (2) cortical thickness in motor and cognitive areas that was negatively related to pain intensity, orbitofrontal cortical thickness that was negatively correlated to pain unpleasantness, and thalamic GM volume correlated to TMD duration, (3) an abnormal relationship between neuroticism and orbitofrontal cortical thickness, (4) abnormal GM aging in nociceptive, modulatory and motor areas, (5) widespread abnormalities in white matter tracts in the brain related to sensory, motor and cognitive functions, (6) reduced trigeminal nerve integrity related to pain duration, and (7) abnormal connectivity in cognitive and modulatory brain regions. In sum, this thesis demonstrates for the first time abnormalities in both peripheral nerve and CNS in patients with TMD.

Temporomandibular disorders (TMD) are a family of prevalent chronic pain disorders affecting masticatory muscles and/or the temporomandibular joint. There is no unequivocally recognized peripheral aetiology for idiopathic TMD. The central nervous system (CNS) may initiate and/or maintain the pain in idiopathic TMD due to sustained or long-term nociceptive input that induces maladaptive brain plasticity, and/or to inherent personality-related factors that may reduce the brain's capacity to modulate nociceptive activity. The main aim of this thesis is to determine whether there are structural neural abnormalities in patients with TMD, and whether these abnormalities are related to TMD pain characteristics, or to neuroticism. The specific aims are to delineate in TMD: (1) gray matter (GM) brain abnormalities and the contribution of pain and neuroticism to abnormalities; (2) the contribution of abnormal brain GM aging in focal cortical regions associated with nociceptive processes; and (3) abnormalities in brain white matter and trigeminal nerve and the contribution of pain. In groups of 17 female patients with TMD and 17 age- and sex- matched controls, magnetic resonance imaging revealed that patients with TMD had: (1) thicker cortex in the somatosensory, ventrolateral prefrontal and frontal polar cortices than controls, (2) cortical thickness in motor and cognitive areas that was negatively related to pain intensity, orbitofrontal cortical thickness that was negatively correlated to pain unpleasantness, and thalamic GM volume correlated to TMD duration, (3) an abnormal relationship between neuroticism and orbitofrontal cortical thickness, (4) abnormal GM aging in nociceptive, modulatory and motor areas, (5) widespread abnormalities in white matter tracts in the brain related to sensory, motor and cognitive functions, (6) reduced trigeminal nerve integrity related to pain duration, and (7) abnormal connectivity in cognitive and modulatory brain regions. In sum, this thesis demonstrates for the first time abnormalities in both peripheral nerve and CNS in patients with TMD.

L’opossum nait dans un état très immature, mais rampe avec ses membres antérieurs (MA) de l’orifice urogénital de la mère à une tétine, où il s’attache pour poursuivre son développement. Des informations sensorielles sont nécessaires pour guider le nouveau-né vers une tétine et déclencher son attachement. Des expériences précédentes ont montré que le système du trijumeau, dont dépend l’innervation somesthésique du museau, influence les mouvements précoces des MA. Le présent projet vise à déterminer si les mécanorécepteurs faciaux sont fonctionnels et exercent une influence sur les MA. On s’intéresse particulièrement aux cellules de Merkel, un mécanorécepteur épidermique innervé par des fibres à adaptation lente de type I (SA I). Ces cellules ont été localisées sur le pourtour du museau de l’opossum nouveau-né en utilisant un traceur cellulaire, l’AM1-43. Nous avons analysé les réponses musculaires des MA consécutives à l’application de forces calibrées au museau sur des préparations in vitro. Ces réponses sont bilatérales et simultanées, très variables, et leur intensité augmente avec la force de la stimulation. Lors de stimulations répétitives pendant 60 min, les réponses diminuent avec le temps. Le retrait de la peau faciale abolit presque ces réponses. De plus, l’application d’un antagoniste des récepteurs métabotropiques du glutamate, qui affecte l’activité des fibres SA I, ou d’un antagoniste des récepteurs purinergiques les diminue fortement, suggérant une participation des cellules de Merkel. Ces résultats soutiennent que le sens du toucher facial relayé par le système du trijumeau est fonctionnel chez l’opossum nouveau-né et qu’il pourrait influencer les mouvements des MA. / The opossum, Monodelphis domestica, is born very immature but crawls, unaided, with its forelimbs (FL) from the mother's birth canal to a nipple where it attaches to pursue its development. Sensory clues are needed to guide the newborn to the nipple and trigger its attachment to it. We postulated that the trigeminal system, responsible for sensory innervation of the face, is involved. Indeed, light pressure applied on the snout evokes FL movements in vivo, low intensity electrical stimulation of the trigeminal ganglion induces motor responses of the FL in vitro, and trigeminal fibers is distributed in the facial dermis and basal epidermis of the newborn. Also, slowly adapting mechanosensory receptors Merkel cells (AM1-43 positive) are present in the face epidermis. To determine if Merkel cells exert an influence on locomotion of newborn opossums, we analyzed the FL muscles responses following application of calibrated forces on the snout in in vitro preparations. Pressure applied to the face induced bilateral and simultaneous FL motor responses, which intensity is proportional to stimulation force. Following consecutive stimulations during 60 min, the responses tended to decrease. Removing the facial skin nearly abolished the responses. Bath applications of the glutamate metabotropic receptor antagonist YM298198, and of the purinergic receptors (P2) antagonist PPADS, decreased the muscles responses. These results support that touch sensitivity of the snout relayed by the trigeminal system is functional in newborn opossums and may influence FL movement, possibly contributing to guiding the animal to the nipple.

La communication entre les neurones est fondée sur leur capacité à changer leur patron de décharge pour l’encodage de différents messages. Pour plusieurs fonctions vitales, comme la respiration et la mastication, les neurones doivent pouvoir générer des patrons d’activité répétitifs, et les groupes de neurones responsables de ces décharges rythmiques sont des générateurs de patron central (GPC). En dépit de recherches soutenues, les mécanismes précis qui sous-tendent la rythmogénèse dans les GPCs ne sont pas bien définis. Le plus souvent, la potentielle contribution des astrocytes demeure grandement inexplorée, même si ces cellules sont aujourd’hui connues pour leur implication dans la modulation synaptique neuronale. Pour nos travaux, le noyau sensoriel principal du trijumeau (NVsnpr) a été pris comme modèle à cause de son rôle central dans les mouvements rythmiques de la mastication. Dans ce noyau, des travaux antérieurs ont montré que la décharge en bouffées rythmiques est déclenchée dans les neurones lorsque la concentration de calcium extracellulaire ([Ca2+]e) est artificiellement baissée. Nous fondant sur cette observation, notre première hypothèse a postulé que la baisse de la [Ca2+]e pouvait survenir de façon physiologique en lien avec des stimulations sensorielles pertinentes. Deuxièmement, parce que les astrocytes ont été impliqués dans le tamponnage et l’homéostasie d’ions extracellulaires comme le K+, nous avons postulé que ces cellules pouvaient jouer un rôle équivalent dans le contrôle de la [Ca2+]e. Nos résultats montrent que les astrocytes peuvent réguler la [Ca2+]e et ainsi contrôler la capacité des neurones à changer leur patron de décharge. Premièrement, en stimulant les afférences sensorielles au NVsnpr, nous avons montré que des baisses physiologiques de la [Ca2+]e sont observées en parallèle à l’apparition de bouffées rythmiques neuronales. Deuxièmement, nous avons démontré que les astrocytes répondent aux mêmes stimuli qui induisent l’activité rythmique neuronale, et que leur blocage avec un chélateur de Ca2+ empêche les neurones de générer un patron de décharge en bouffées rythmiques. Cette habilité est rétablie en rajoutant la S100β, une protéine astrocytaire liant le Ca2+, dans le milieu extracellulaire, alors que l’anticorps anti-S100β empêche l’activité rythmique. Ces résultats indiquent que les astrocytes régulent une propriété neuronale fondamentale : la capacité à changer de patron de décharge. Ainsi, les GPCs dépendraient des fonctions intégrées des astrocytes et des neurones. Ces découvertes pourraient avoir des implications transposables à plusieurs autres circuits neuronaux dont la fonction dépend de l’induction d’activité rythmique. / Communication between neurons rests on their capacity to change their firing pattern to encode different messages. For several vital functions, such as respiration and mastication, neurons need to generate a repetitive firing pattern, and the groups of neurons responsible for these rhythmic discharges are called central pattern generator (CPG). Despite intense research in this field, the exact mechanisms underlying rhythmogenesis in CPGs are not completely defined. In most instances, the potential contribution of astrocytes is largely unexplored, even though these cells are now well known to be involved in neuronal synaptic modulation. In our work, the trigeminal main sensory nucleus (NVsnpr) was used as a model owing to its central role in the rhythmic movement of mastication. Previous work have shown that rhythmic bursting discharge is triggered in NVsnpr neurons when extracellular calcium concentration ([Ca2+]e) is artificially decreased. Based on this observation, our first hypothesis postulated that the reduction of [Ca2+]e could also happen physiologically in relation to relevant sensory stimulation. Secondly, because astrocytes have been involved in the buffering and the homeostasis of extracellular ions like potassium, we have postulated that these cells could also play a role in the control of [Ca2+]e. The results presented in this thesis show that astrocytes can regulate [Ca2+]e and thus control the ability of neurons to change their firing pattern. First, we showed that stimulation of sensory afferent fibers to the NVsnpr induced neuronal rhythmic bursting and in parallel reduction of [Ca2+]e . Secondly, we have demonstrated that astrocytes respond to the same sensory stimuli that induce neuronal rhythmic activity, and their blockade with a Ca2+ chelator prevents generation of neuronal rhythmic bursting. This ability is restored by adding S100β, an astrocytic Ca2+-binding protein, to the extracellular space, while the application of an anti- S100β antibody prevents generation of rhythmic activity. These results indicate that astrocytes regulate a fundamental neuronal property: that is the capacity to change their firing pattern. Thus, CPG functions result from integrated neuronal and glial activities. These findings may have broad implications for many other neural networks whose functions depend on the generation of rhythmic activity.

Esse trabalho teve como objetivos avaliar as características odontologicas, verificar a ocorrência de complicações sensitivas orofaciais e determinar os aspectos funcionais mandibulares de 105 doentes com neuralgia idiopática do trigêmeo (NIT) submetidos \'a compressão radiculo-ganglionar com balão inflável. Foram realizadas 5 avaliações para cada doente: uma pré-cirúrgica e 4 pós-cirúrgicas (7, 30, 120 e 210 dias). Ocorreu comprometimento sensitivo mais intenso no território dos ramos maxilar e mandibular do nervo trigêmeo (p < 0,001) e poucas anormalidades no território do ramo oftálmico (p = 0,1815). As qualidades sensitivas calor, frio, tato, e dor foram afetadas. As queixas subjetivas de dormência foram mais frequentes do que as objetivadas durante o exame de sensibilidade facial (p < 0,001). foi elevada a ocorrência de disfunção oclusal (62,9%); 42,6% dos doentes apresentaram queixas espontâneas relacionadas à mastigação; houve intensificação da dor miofascial na musculatura mastigatória após a cirurgia (p < 0,001), que retornou aos valores iniciais após os 210 dias; a mobilidade mandibular também agravou-se (p < 0,001). Os autores concluiram que o procedimento é eficaz e seguro quando aplicado em doentes com NIT envolvendo o ramo oftálmico. Entretanto, recidiva é frequente e as complicações sensitivas e ou motoras orofaciais poderiam comprometer a qualidade de vida e dificultar a reabilitação funcional dos doentes. / The aim of this study was to determine dental characteristics, abnormalities in masticatory function and ocurrence of orofacial sensorial complications in 105 patients with trigeminal neuralgia treated with radiculo-ganglionar compression of the trigeminal ganglion with balloon. The patients were evaluated in the pre-operative period and in 4 post-operative evaluations (7, 30, 120 and 210 days). Sensory deficits were more severe in the area innervated by the maxillary and the mandibular trigeminal branches (p < 0.001); the ophthalmic branch presented abnormalities in few cases. The sensory qualities heat, cold, tactile and pain were affected. Subjective numbness was more frequent than sensory abnormalities findings at the post-operative sensitive evaluation (p < 0.001). Dental occlusion abnormalities were observed in 62.9% of the patients and 42.6% of patients\' complaints were spontaneous masticatory difficulties. During the post-operative period, myofascial pain of the masticatory muscles was statistically significant (p < 0.001), but normalized after 210 days in average. It also compromised the jaw mobility (p < 0.001). It was concluded that this procedure is safe for patients with idiopathic trigeminal neuralgia involving the ophthalmic branch; however, sensory and motor complications of the method can affect the quality of life and rehabilitation of the patient.

Neuralgia trigeminal (NT) é um tipo de dor neuropática orofacial intensa e debilitante sentida ao longo do nervo trigêmeo. Seu diagnóstico é meramente clínico, sem exames específicos, o que dificulta a presteza e eficácia do tratamento. O sintoma mais característico é alodínia mecânica relacionada à sensibilização central. Seu tratamento é farmacológico e/ou cirúrgico, no entanto pode trazer poucos benefícios e/ou importantes efeitos colaterais sem a garantia de remissão total dos sintomas. No tratamento farmacológico, os anticonvulsivantes são fármacos de primeira linha de tratamento, mas também são obtidas respostas de melhora com o uso de fármacos ansiolíticos e opioides. A dificuldade no tratamento da NT se deve, em parte, à falta de compreensão dos mecanismos envolvidos na geração da dor; sendo, desta forma, imprescindível a busca de um melhor entendimento da fisiopatologia da NT buscando identificar as vias envolvidas no processo de dor. Sendo assim, o objetivo deste estudo foi avaliar o possível envolvimento das vias gabaérgica, glutamatérgica e opioidérgica na fisiopatogenia de um modelo de neuralgia trigeminal em ratos. A resposta nociceptiva dos animais foi avaliada por meio do teste de von Frey eletrônicoantes e após a administração de agonistas gabaérgicos e opioidérgico e de antagonista glutamatérgico. Além da a resposta nociceptiva, foram avaliados os níveis de fator neurotrófico derivado do cérebro (BDNF) em gânglio trigeminal, tronco encefálico e córtex pré-frontal. Todos os procedimentos experimentais foram realizados na Unidade de Experimentação Animal (UEA), Unidade de Análises Moleculares e de Proteínas (UAMP) do Hospital de Clínicas de Porto Alegre (HCPA). A indução da dor neuropática foi por constrição crônica do nervo infraorbitário (CCI-ION) proposto por Imamura em 1997 adaptado (Imamura Y., Kawamoto H., Nakanishi O. Charactherization of heat-hiperalgesia in experimental trigeminal neuropathy in rats. Exp Brain Res, 1997; 116: 97-103.). Para isso foram utilizados 112 ratos Wistar, machos divididos para o procedimento cirúrgico em 3 grupos: Controle Total, Dor e Sham. Quatorze dias após o procedimento cirúrgico, tempo necessário para o desenvolvimento da dor neuropática, realizamos dois protocolos e dividimos os grupos em: protocolo 1: controle veículo, cirurgia sham veículo, cirurgia sham agonista benzodiazepínico, cirurgia sham antagonista glutamatérgico, dor veículo, dor agonista benzodiazepínico, dor antagonista glutamatérgico; protocolo 2: controle veículo, cirurgia sham veículo, cirurgia sham agonista gabaérgico, cirurgia sham agonista opioidérgico, dor veículo, dor agonista gabaérgico, dor agonista opioidérgico. Os animais dos grupos sham sofreram apenas incisão cirúrgica, sem constrição do nervo. Durante o procedimento cirúrgico a indução anestésica foi feita com cetamina (50mg/kg) e xilazina (5mg/kg) e a manutenção, com isoflurano 2-3%. O controle da dor pós-operatória foi obtido com cloridrato de tramadol (10mg/kg) no intervalo de 12h por 48h. A resposta hiperalgésica mecânica (von Frey) foi avaliada antes da indução do modelo (medida basal), 7 e 14 dias após cirurgia, pré-administração dos fármacos, 15,30 e 60 minutos após administração dos fármacos: veículo - solução salina; agonista benzodiazepínico – diazepam (2 mg/kg); antagonista glutamatérgico - MK- 801 (0,25mg/Kg), agonista opioidérgico – morfina (5,0 mg/kg) e agonista gabaérgico – fenobarbital (100mg/kg). Os dados foram analisados por Equações Estimativas Generalizadas e expressos em média+EPM. Todos os procedimentos foram aprovados pelo CEUA/HCPA e CEUA/UFRGS sob número: 14-0604 e 29310, respectivamente. Inicialmente foram realizados testes para verificação do modelo de dor, foi observado que 14 dias após cirurgia de constrição do nervo infraorbitário, os animais do grupo dor apresentaram redução nos limiares de retirada da face no teste de von Frey facial comparado aos grupos controle e sham [GEE: interação grupo x tempo (Waldχ2=15,81; 2, P<0,05)]. Após o estabelecimento do modelo, foram realizados testes nociceptivos para avaliação das vias envolvidas na neuralgia trigeminal, observou-se interação grupo x tempo (Wald _2=175,74;18, P<0,01). No protocolo 1, os animais do grupo dor que receberam agonista benzodiazepínico (diazepam) apresentaram aumento no limiar nociceptivo a partir de 15 minutos após administração e este resultado permaneceu por até 60 minutos. Os animais do grupo dor que receberam antagonista glutamatérgico (MK-801) apresentaram aumento no limiar de retirada da face somente na avaliação realizada 60 minutos após a administração do fármaco. No protocolo 2, análise estatística mostrou interação entre as variáveis tempo x grupo (Wald χ2=657,53;18, P<0,01), os animais do grupo dor que receberam fenobarbital apresentaram aumento no limiar nociceptivo a partir de 15 minutos após administração e este resultado permaneceu por até 60 minutos. Por outro lado, os animais que receberam morfina apresentaram um aumento no limiar nociceptivo significativamente maior que os animais que receberam fenobarbital, no entanto ambos os fármacos reverteram a hiperalgesia induzida pela exposição ao modelo de dor. Na análise bioquímica, em córtex e tronco encefálico foi observado aumento nos níveis de BDNF tanto nos animais submetidos ao modelo de NT quanto aos que foram expostos a cirurgia sham (ANOVA de uma via/SNK, F(2,22)=13,46; F(2,25)=6,08, P<0,05, respectivamente. Em nível periférico foi observado aumento nos níveis de BDNF em gânglio trigeminal nos animais submetidos ao modelo de NT (ANOVA de uma via/SNK,F(2,22)=4,09; P<0,05). Em soro não foi observada diferença nos níveis de BDNF entre os grupos (ANOVA de uma via, P>0,05). Com base nos resultados encontrados, este estudo sugere que há o envolvimento das vias gabaérgica, glutamatérgica e opioidergica no processamento da dor neuropática orofacial (neuralgia trigeminal) sugerindo que há funcionalidade dos receptores testados. No entanto, estes agonistas gabaérgicos não induziram analgesia nos animais controles e cirurgia sham sugerindo uma alteração na liberação ou na síntese de GABA nos animais com NT. Neste estudo, não foram realizadas análises de expressão e quantificação de receptores, apenas avaliou-se a resposta nociceptiva e os níveis de BDNF centrais e periféricos. De acordo com estes achados a via gabaérgica teve uma resposta mais rápida que a via glutamatérgica. Alguns estudos demonstram que os receptores glutamatérgicos estão localizados em fibras não mielinizadas na periferia isto pode justificar o atraso na resposta dos animais que receberam antagonista glutamatérgico em nosso estudo. Quanto aos níveis de BDNF observamos que há um aumento desta neurotrofina nos animais submetidos ao modelo de NT sugerindo um envolvimento deste neuromodulador em eventos neuroplásticos da NT. / Trigeminal neuralgia (TN) is a type of intense and debilitating orofacial neuropathic pain felt along the trigeminal nerve. The diagnosis is purely clinical, without specific tests, which makes the promptness and effectiveness of treatment. The most characteristic symptom is mechanical allodynia related to central sensitization. Its treatment is pharmacological and / or surgical treatment, however can bring few benefits and / or significant side effects without the guarantee of complete remission of symptoms. In drug treatment, the anticonvulsant drug are first-line treatment, but also enhances responses are obtained with the use of anxiolytic drugs and opioids. The difficulty in treating NT is due in part to the lack of understanding of the mechanisms involved in the generation of pain; It is thus essential to search for a better understanding of the pathophysiology NT seeking to identify the pathways involved in pain process. Thus, the aim of this study was to evaluate the possible involvement of GABAergic pathways, glutamatergic and opioidergic in the pathogenesis of a model of trigeminal neuralgia in rats. The nociceptive response of the animals was assessed using the von Frey test eletrônicoantes and after administration of opioid agonists and GABAergic and glutamatergic antagonist. In the nociceptive response was evaluated derived neurotrophic factor levels in the brain (BDNF) in the trigeminal ganglia, brainstem, and prefrontal cortex. All experimental procedures were performed in the Animal Experimentation Unit (UEA), Unit of Analysis and Molecular Protein (UAMP) Porto Alegre Clinical Hospital (HCPA). The induction of neuropathic pain was by chronic constriction of the infraorbital nerve (CCI-ION) proposed by Imamura in 1997 adapted. For that they were used 112 male Wistar rats divided into the surgical procedure into 3 groups: Full Control, Pain and Sham. Fourteen days after surgery, time required for the development of neuropathic pain, performed two protocols and divided groups: Protocol 1: vehicle control, sham surgery vehicle, sham surgery benzodiazepine agonist, sham surgery glutamatergic antagonist, vehicle pain, agonist benzodiazepine pain and antagonist glutamatergic pain. Protocol 2: vehicle control, vehicle sham surgery, sham surgery gabaergic agonist, sham surgery opioid agonist, vehicle pain, gabaergic agonist pain, opioid agonist pain. The animals of the sham group underwent only surgical incision without nerve constriction. During surgery anesthesia was induced with ketamine (50 mg / kg) and xylazine (5 mg / kg) and maintained with isoflurane 2-3%. The control of postoperative pain has been obtained with tramadol hydrochloride (10mg / kg) at 12h intervals for 48 hours. The mechanical hyperalgesic response (von Frey) was assessed before induction model (baseline measurement), 7 and 14 days after surgery, pre-administration of drugs, 15,30 and 60 minutes after drug administration: Vehicle - saline; benzodiazepine agonist - diazepam (2 mg / kg); glutamatergic antagonist - MK-801 (0.25mg / kg), opioid agonist - Morphine (5.0 mg / kg) and gabaergic agonist - phenobarbital (100 mg / kg). Data were analyzed by Equation Generalised Estimates and expressed as mean ± SEM. All procedures were approved by CEUA / HCPA and CEUA / UFRGS under number: 14-0604 and 29310, respectively. Initially tests were carried out to verify the pain model, it was observed that 14 days after infraorbital nerve constriction surgery, animals pain group showed a reduction in the face withdrawal thresholds to von Frey facial test compared to control and sham groups [ GEE: group x time interaction (Wald 2 = 15.81; 2, P <0.05)]. After the model category, nociceptive tests were performed to assess the pathways involved in the trigeminal neuralgia, observed group x time interaction (Wald _2 = 175.74; 18 P <0.01). In protocol 1, the animals in the group who received pain benzodiazepine agonist (diazepam) showed an increase in nociceptive threshold from 15 minutes after administration and this result remained for up to 60 minutes. Animals of group pain that received glutamate antagonist (MK-801) showed an increase in the withdrawal threshold of the face only in the assessment was performed 60 minutes after drug administration. In protocol 2, statistical analysis showed interaction between the variables time x group (Wald χ2 = 657.53; 18, P <0.01), the animals showed increased nociceptive threshold from 15 minutes after administration of phenobarbital and morphine and this result remained for up to 60 minutes. On the other hand, animals receiving morphine showed an increased nociceptive threshold significantly higher in the animals receiving phenobarbital, however both drugs reversed hyperalgesia induced by exposure to pain model. In biochemical analysis, cortex and brainstem was observed increase in BDNF levels in both animals subjected to NT model and those who were exposed to sham surgery (one-way ANOVA / SNK, F (2,22) = 13.46 ;. F (2,25) = 6.08, P <0.05, respectively, at the peripheral level increase was observed in BDNF levels in trigeminal ganglion of animals subjected to the NT model (one-way ANOVA / SNK F ( 2,22) = 4.09; P <. 0.05). In serum was no difference in BDNF levels between groups (one-way ANOVA, P> 0.05) based on the results, this study. suggests that there is involvement of gabaergic pathways, glutamatergic and opioidergic processing orofacial neuropathic pain (trigeminal neuralgia) suggesting that there is functionality of the tested receptors. However, these GABAergic agonists did not induce analgesia in control animals and sham surgery suggesting a change in release or GABA synthesis in animals with NT. in this study, were not realized expression analysis and quantification of receptors, only evaluated the nociceptive response and the central and peripheral levels of BDNF. According to these findings, gabaergic pathways had a faster response than glutamatergic pathways. Some studies have shown that glutamate receptors are located in non-myelinated fibers in the periphery that can justify the delay in the animals that received glutamatergic antagonist in our study. As for BDNF levels we observed that there is an increase of this neurotrophin in animals subjected to NT model suggesting an involvement of this neuromodulator in neuroplastic events NT.

A deeper understanding of both movement control and the effects of nociceptor inputs on our motor systems is critical for proper clinical diagnosis of musculo-skeletal dysfunctions and for development of novel rehabilitation schemes. In the jaw system, masticatory movements are produced by a central pattern generator (CPG) located in the brainstem. Considerable efforts have been made in deciphering this neuronal network. The present thesis contributes towards an increasingly detailed understanding of its essential elements, and presents a hypothesis of how deep somatic pain (i.e. muscle pain) may be evoked and interferes with the masticatory CPG circuitry. In Paper I, the expression of c-Fos-like protein was used as a molecular marker to visualize brainstem neurons that were active during induced fictive mastication in the anesthetized and paralyzed rabbit. Our findings provide a previously lacking detailed record of the neuronal populations that form the masticatory motor pattern. Certain cells were located in brainstem areas previously suggested to be involved in the masticatory CPG. However, it was a new finding that neurons in the dorsal part of the trigeminal main sensory nucleus (NVsnpr-d) may belong to this circuitry. Paper II focused on the discovered neurons in NVsnpr in an in vitro slice preparation from young rats.  Intracellular recordings allowed us to define two cell types based on their response to depolarizing current. Microstimulation applied to the trigeminal motor nucleus, its reticular border, the parvocellular reticular formation and the nucleus reticularis pontis caudalis, elicited postsynaptic potentials in 81% of the neurons tested. Responses obtained were predominately excitatory and sensitive to gluta-matergic antagonists DNQX or/and APV. Some inhibitory and biphasic responses were also evoked. Bicuculline methiodide or strychnine blocked the IPSPs indicating that they were mediated by GABAA or glycinergic receptors. About one third of the stimulations activated both types of neurons antidromically. Neurons in NVsnpr-d seem to gather all the conditions that can theoretically account for a role in masticatory rhythm generation. In Paper III, the masticatory model system was used to investigate the possible role of muscle spindle primary afferents in development of persistent musculoskeletal pain. Following intramuscular acidic (pH 4.0) saline injections of rat masseter muscles, in vitro whole cell recordings were done from jaw closing muscle spindle somata located in the trigeminal mesencephalic nucleus (NVmes). Compared to control neurons, the somata of afferents exposed to acid had more hyperpolarized membrane potentials, more hyperpolarized thresholds for firing, high frequency membrane oscillations and ectopic bursting of action potentials. These changes in membrane properties lasted for up to 35 days. Within the same time frame experi-mental animals showed hypersensitivity to touch on the skin covering the injected muscle. Similar saline injections also resulted in a significant increase of activity dependent c-Fos expression in NVmes neurons compared to controls. Immuno-fluorescence and lectin binding studies indicated that small-caliber muscle afferents containing known nociceptor markers (CGRP, SP, P2X3, TRPV1 and IB4) and expressing glutamate receptors are found close to the annulo-spiral endings of the NVmes afferents. Combined, our new observations support the hypothesis that excessive release of glutamate, within muscle spindles due to ectopically evoked antidromic action potentials, could lead to development of persistent musculoskeletal pain by activation and/ or sensitization of adjacent muscle afferent nociceptors.

The controlled lowering of core body temperature to 32°C to 34°C is defined as therapeutic hypothermia (TH). Therapeutic hypothermia has been shown to improve neurological outcome and survival in unconscious patients successfully resuscitated after cardiac arrest. Brain temperature is important for cerebral protection therefore methods for primarily cooling the brain have also been explored. This thesis focuses on the likelihood that intranasal cooling can induce, maintain and control cerebral hypothermia. The method uses bilaterally introduced intranasal balloons circulated with cold saline. Selective brain cooling induced with this method was effectively accomplished in pigs with normal circulation while no major disturbances in systemic circulation or physiological variables were recorded. The temperature gradients between brain and body could be maintained for at least six hours. Intranasal balloon catheters were used for therapeutic hypothermia initiation and maintenance during and after successful resuscitation in pigs. Temperature reduction was also obtained by combined intranasal cooling and intravenous ice-cold fluids with possible additional benefits in terms of physiologic stability after cardiac arrest. Rewarming was possible via the intranasal balloons. In these studies brain temperature was recorded invasively by temperature probes inserted in the brain. The fast changes in pig’s brain temperature could also be tracked by a non-invasive method. High-spatial resolution magnetic resonance spectroscopic imaging (MRSI) without internal reference showed a good association with direct invasive temperature monitoring. In addition the mapping of temperature changes during brain cooling was also possible. In awake and unsedated volunteers subjected to intranasal cooling brain temperature changes were followed by two MR techniques. Brain cooling was shown by the previously calibrated high-spatial resolution MRSI and by the phase-mapping method. Intranasal cooling reduced body temperature slightly. The volunteers remained alert during cooling, the physiological parameters stable, and no shivering was reported.

Le mouvement masticatoire est généré et coordonné par un générateur de patron central (GPC) situé au niveau du pont. Plusieurs résultats antérieurs de notre laboratoire soutiennent que le réseau de neurones à l’origine de la rythmogénèse est situé dans le noyau sensoriel principal du nerf trijumeau (NVsnpr). Ces mêmes expériences révèlent que des diminutions de la concentration calcique extracellulaire ([Ca2+]e) tiennent une place importante dans la génération des bouffées de décharges des neurones de cette région. Notre laboratoire tente de vérifier si la contribution des astrocytes à l’homéostasie de la concentration calcique extracellulaire est impliquée dans la genèse du rythme. Cette étude a pour but la caractérisation spatiale du syncytium astrocytaire au sein du NVsnpr dorsal et l’étude de l’effet de la [Ca2+]e sur les propriétés astrocytaires électrophysiologiques et de connectivité. Nous avons utilisés pour ce faire la technique d’enregistrement par patch-clamp sur une préparation en tranche de tronc cérébral de rat. Nous démontrons ici que la diminution de la [Ca2+]e n’affecte pas les propriétés électrophysiologiques astrocytaires, mais induit une augmentation de la taille du syncytium. De plus, nous établissons l’existence au sein du NVsnpr dorsal d’une organisation anatomofonctionnelle du réseau astrocytaire calquée sur l’organisation neuronale. / The masticatory movement is generated and coordinated by a central pattern generator (CPG) located in the pons. Previous results from our laboratory suggest that the neural network responsible for its rythmogenesis is located in the trigeminal main sensory nucleus (NVsnpr). Moreover, results indicate that in this region, decrease in extracellular calcium concentration ([Ca2+]e) plays an important role in genarating burst. One of our laboratory's goal is to assess if the contribution of astrocytes to the extracellular calcium concentration homeostasis is involved in the genesis of the mastication rhythm. With this study, we characterized the astrocyte syncytium within the NVsnpr and measured the effect of [Ca2+]e on the astrocytes electrophysiology and their networks. A patch-clamp recording technique in conjunction with a rat brain stem slice preparation was used. We demonstrate that a decrease in [Ca2+]e does not affect the electrophysiological properties of astrocytes but induces an increase in the size of the syncytium. We also report the existence, within the dorsal NVsnpr, of an anatomofunctional organization between neurons and astrocytes.

No description available.