Existem crescentes evidências sobre a contribuição da liberação de noradrenalina (NA) central na consolidação das memórias. Teoricamente, o Núcleo do Trato Solitário (NTS) recebe informações e diversos estímulos periféricos, que são então projetados ao Núcleo Paragigantocelular (PGi). Este, por sua vez, utiliza neurotransmissores, predominantemente excitatórios, para influenciar a ativação do Locus Coeruleus (LC). Então, o LC envia projeções noradrenérgicas ao hipocampo e à amígdala, influenciando os processos mnemônicos. Aqui nós demonstramos que a inibição pelo muscimol do NTS, PGi ou LC até 3 horas após o treino na tarefa de reconhecimento de objetos (RO) impede a consolidação da memória medida 24 h após o treino. Adicionalmente, a infusão de timolol, um antagonista de receptores β-adrenérgicos, na região CA1 do hipocampo também impede a consolidação deste tipo de memória. A infusão de NA na região CA1 do hipocampo não altera a retenção da memória, mas, reverte o prejuízo causado pela inibição do NTS, PGi ou LC. A infusão de NMDA no LC após a inibição do NTS ou PGi também reverte essa amnésia. Concomitantemente, verificamos que a inibição NTS, PGi ou LC bloqueia o aumento da expressão do fator neurotrófico derivado do cérebro (BDNF, do inglês brain-derived neurotrophic factor) que ocorre 120 min após o treino na tarefa de reconhecimento de objetos na região CA1 do hipocampo. Também a infusão de NA na região CA1 do hipocampo após a inibição do NTS, PGi ou LC ou de NMDA no LC após a inibição do NTS ou PGi promovem novamente o aumento do BDNF120 min após o treino no RO. Com isso conclui-se que a ativação da via NTS-PGi-LC-Hipocampo é necessária para que ocorra consolidação da memória de RO, na qual desempenha um papel o BDNF hipocampal. / There is evidence of the contribution of brain noradrenaline release (NA) to memory consolidation. The Nucleus of the Solitary Tract (NTS) receives information originated by peripheral stimuli and projects to the Paragigantocellularis Nucleus (PGi), which influences the Locus Coeruleus (LC) through excitatory neurotransmitters. The LC sends noradrenergic projections to the hippocampus and amygdala, influencing the memory processes. Here we show that inhibition by muscimol of NTS, PGi or LC up to 3 h after object recognition training impairs the consolidation of the memory measured 24 h later. Additionally, the infusion of timolol in the CA1 region of hippocampus also inhibits consolidation of this type of memory. The infusion of NA into the CA1 region of hippocampus does not alter memory consolidation of this task, but reverts the deleterious effect of NTS, PGi or LC inhibition. The infusion of NMDA in LC after inhibition of NTS or PGi also reverts the amnesia. Concomitantly, the inhibition of NTS, PGi or LC blocks the increase of brain-derived neurotrophic factor (BDNF) expression in CA1 that occurs 120 min after training in the object recognition task. Further, the infusion of NA in CA1 after inhibition of NTS, PGi or LC; or of NMDA in LC after inhibition of NTS or PGi promotes the BDNF increase seen 120 min after object recognition training. Thus, it is concluded that the activation of NTSPGi- LC-Hippocampus pathway is necessary for consolidation of the object recognition memory, and hippocampal BDNF is involved in this process.

Objetivos: Este trabalho teve como objetivos avaliar in vivo os efeitos microcirculatórios e a sobrevida de animais submetidos ao choque hemorrágico tratados com vasopressina e noradrenalina associadas à reposição volêmica com solução de NaCl 0,9% . Desenho do estudo: Estudo prospectivo, randomizado, controlado, intervencionista em modelo animal. Materiais e métodos: Utilizou-se hamsters machos do tipo sírio dourado, com idade entre 6 e 8 semanas e massa corporal entre 60 e 80 gramas. Os animais foram anestesiados para colocação de uma câmara dorsal. Após 5 a 7 dias, foram re-anestesiados para implante de cânulas na carótida e na veia jugular. No dia seguinte realizou-se o experimento. Os animais sofreram choque hemorrágico por meio da retirada de 40% da volemia, definida como 7% do peso corporal, e mantidos em choque por uma hora. Após, os animais foram aleatoriamente divididos em três grupos : Grupo SF0,9% (N = 6) - recebeu solução de NaCl 0,9% em volume de duas vezes o volume de sangue retirado; Grupo VP recebeu solução de NaCl 0,9% em volume de duas vezes o volume de sangue retirado, associado à infusão contínua durante uma hora de vasopresssina (0,0001 UI/kg/min por uma hora); e Grupo Nora. recebeu solução de NaCl 0,9% em volume de duas vezes o volume de sangue retirado, associado à infusão contínua de solução de noradrenalina (2 g/kg/min por uma hora). Foram avaliados os diâmetros das arteriolas e vênulas e a densidade capilar funcional (DCF) no momento basal, após o choque e após o tratamento. Os parâmetros laboratoriais observados foram: pH, HCO-3, BEx, paO2, paCO2 e lactato durante as três fases do experimento. Após o término do tratamento, foi visualizado o rolamento e adesão de leucócitos , assim como a sobrevida dos animais durante setenta e duas horas. Resultados: A terapia com reposição volêmica por si ou associada à vasopressina ou à noradrenalina não alterou valores relativos à gasometria arterial ou lactato em relação ao choque. A terapia com vasopressina associada à reposição volêmica manteve a densidade capilar funcional após ressuscitação do choque hemorrágico, (97% do valor basal da mediana), enquanto tratamento com solução de NaCl 0,9% apenas, não obteve o mesmo resultado (70% do valor basal da mediana). A noradrenalina associada à reposição volêmica piorou a densidade capilar funcional após o tratamento (44% do valor basal da mediana). A sobrevivência em setenta e duas horas foi significativamente menor no grupo da noradrenalina do que no grupo da vasopressina (33% em relação ao grupo Vaso). Ao final do experimento não foi observada diferença estatisticamente significativa relativa à adesão ou rolamento de leucócitos. Conclusão: Durante o choque hemorrágico, o tratamento com infusão de solução de NaCl 0,9% associada à vasopressina mantém a DCF, enquanto que o tratamento com solução de NaCl 0,9% somente ou associada à noradrenalina pioram a DCF. O tratamento com vasopressina melhora a sobrevida dos animais, em comparação ao tratamento com noradrenalina. Embora a adesão leucocitária não esteja significativamente alterada entre os grupos, houve uma tendência em obtermos uma adesão menor no grupo da vasopressina. / Objectives: The goal of this work was to evaluate in vivo the microcirculatory effects and survival of animals subjected to hemorrhagic shock treated with vasopressin or noradrenalin associated to volume infusion associated with NaCl 0,9% Study design: Prospective, randomized, controlled, intervencionist study in animal model. Materials and methods: Golden Syrian hamsters were used, aging between 6 and 8 weeks with body mass ranging from 60 to 80 grams. Animals were anesthetized for dorsal chamber implant. After 5 to 7 days there was a new anesthesia for carotid artery and jugular vein catheter implantation. Next day the experiment took place. Animals suffered a hemorrhagic shock by withdrawal of 40% of blood volume, defined as 7% of body weight, and kept in shock condition for 1 hour. After, animals were randomly divided in three groups. SF0,9% group (N=6) received NaCl 0,9% two times the shed volume; VP group (N=6) received NaCl 0,9% two times the shed volume plus continuous infusion for one hour of vasopressin solution (0,0001UI/kg/min for one hour); Nora group received NaCl 0,9% two times the shed volume plus continuous infusion of noradrenalin solution (2mcg/kg/min for one hour). Arteriolar diameter, venular diameter and functional capillary density (FCD) were evaluated in baseline, after shock and after treatment. Laboratory parameters observed were: pH, HCO-3, BEx, paO2, paCO2 and lactate during all three phases of experiment. After end of treatment, leucocyte rolling and adhesion were visualized, as well as animal survival during seventy two hours. Results: Volume infusion by itself or associated with vasopressin or noradrenalin didnt altered blood gas analysis values or lactate related to hemorrhagic shock. Vasopressin therapy associated with volume infusion sustained functional capillary density after hemorrhagic shock resuscitation (97% of baseline median values), while treatment with NaCl 0,9% only, didnt obtained the same result (70% of baseline median values). Noradrenalin associated to volume infusion worsened the functional capillary density after treatment (44% of baseline median values). Survival in seventy two hours were significantly lower in noradrenalin group comparing to vasopressin group (33% relating to Vaso group). In the end of experiment wasnt observed any significantly statistical difference relative to leucocyte rolling or adhesion. Conclusion: During hemorrhagic shock, treatment with NaCl 0,9% infusion associated with vasopressin sustains FCD, while treatment with NaCl 0,9% only or associated to noradrenalin worsens FCD. Treatment with vasopressin solution improves survival comparing to noradrenalin infusion treatment. Although leucocyte adhesion wasnt significantly altered among groups, there was a trend in observe lesser adhesion in vasopressin group.

Orientador: Dora Maria Grassi-Kassisse / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-07T08:09:26Z (GMT). No. of bitstreams: 1 Moreira_MariliaFrancisco_M.pdf: 1378785 bytes, checksum: dcaf29a0a3f00f96318d03ee2362989c (MD5) Previous issue date: 2005 / Resumo: Lipídios provenientes da dieta têm um importante efeito no sistema de sinalização transmembrana presente nas células cardíacas. Um vez ingeridos em excesso ocorre aumento no conteúdo de colesterol na membrana da célula cardíaca o qual afeta a atividade da adelilil ciclase ligada ao receptor ß e conseqüentemente as respostas cronotrópicas e inotrópicas às catecolaminas causando arritmogênese. O objetivo desta tese foi analisar os efeitos de sessões de natação com o objetivo de prevenir os efeitos de dieta hiperlipídica sobre a sensibilidade de átrios direitos isolados a agonistas adrenérgicos. Ratos Wistar machos adultos foram usados após uma semana de adaptação em salas climatizadas 22±2ºC e com ciclo claro-escuro de 12 h (luzes acendendo as 6:30 da manhã). Os experimentos foram realizados de acordo com os princípios para utilização de animais em pesquisa e educação e adotado pelo COBEA (Colégio Brasileiro de Experimentação Animal). Os animais foram randomicamente divididos em dois grupos, sedentários (S) e que praticaram exercício (T). O exercício constou de sessões de natação na freqüência de 5 dias na semana com 50 minutos de duração durante 20 dias (4x5 dias) em tanque de água com temperatura de 34 ± 2oC. Estes dois grupos foram ainda subdivididos em 2 subgrupos, o que recebia ração padrão (N) e outro que recebia a dieta rica em lipídios (H). O átrio direito foi isolado e curvas cumulativas dose-resposta a noradrenalina (NA) e isoprenalina (ISO) foram obtidas, na ausência ou presença de inibição da recaptação neuronal e extraneuronal. Não houve alteração na freqüência cardíaca basal ou resposta máxima aos agonistas nos átrios isolados dos diferentes grupos experimentais (NS, NT, HS e HT). O programa de exercício físico proposto induziu em ratos que ingeriram dieta padrão uma subsensibilidade à noradrenalina e uma supersensibilidade a isoprenalina. Os átrios direitos isolados de ratos sedentários que ingeriram dieta rica em lipídios apresentaram, após quatro semanas, subsensibilidade a noradrenalina e nenhuma alteração na sensibilidade a isoprenalina. A associação da dieta rica em lipídios com o programa de exercício físico preveniu as alterações observadas tanto pela dieta hiperlipídica como pelo programa de exercício isoladamente. Os valores pD2 para NA foram: 7,44±0,09 (NS); 6,65±0,17* (HS); 6,52±0,25* (NT); 7,15±0,04**# (HT) e para ISO foram: 8,37±0,12 (NS); 8,52±0,10 (HS); 8,94±0,09* (NT); 8,55±0,08 (HT). Onde as diferenças significativas (p<0.05 ANOVA seguida de Tukey) foram indicadas como segue: * vs NS; ** vs NT, # vs HS. Este programa de exercício físico induziu alterações na resposta atrial semelhantes àquelas observadas no modelo de choques nas patas (três sessões diárias), ou seja, diminuição na resposta mediada pelos adrenoceptores ß1 (NA) e aumento na resposta mediada pelo adrenoceptor ß2 (ISO). A dieta hiperlipídica oferecida aos ratos durante 4 semanas induziu subsensibilidade a noradrenalina e esta resposta é provavelmente devida a alterações na membrana lipídica e alta atividade adrenérgica relatada neste modelo animal. O motivo da ausência de alterações cardíacas quando da associação dieta hiperlipídica e exercício físico precisa sem investigado / Abstract: Dietary lipids has an important effect on transmembrane signaling system in the heart increasing the cardiac membrane cholesterol content, which in time affect ß-adrenoceptor/adenylyl cyclase activity and the inotropic and chronotropic responses to catecholamines causing arrhytmogenesis.The aim of this work was to analyze the effect of swimming, to prevent the effects of high fat-CHO diet on the sensitivity to adrenergic agonists in rat isolated right atria. Adult male Wistar rats were used after one week of adaptation in acclimated room at 22±2ºC and 12h light-dark cycle (lights on at 6:30 a.m.). The experiments were carried out in accordance with the principles for the use of animals in research and education and adopted by COBEA (Brazilian College for Animal Experimentation). The animals were randomly divided into two groups, sedentary (S) and exercised (T) with a swimming sessions, 5 days a week (50 min. session) during 20 days (4x5 days) in a water glass tank with temperature at 34 ± 2oC. These two groups were divided into two subgroups; one of them fed a standard chow (N) and the other, a high fat-CHO diet (H). The atrium was isolated as described before and cumulative concentration-response curves to noradrenaline (NA) and isoprenaline (ISO) were obtained, in the absence or presence of inhibitors of neuronal and extraneuronal uptake. There was no alteration in the basal heart rate or maximal response to the agonists in isolated atria rats from different groups (NS, NT, HS and HT). The physical exercise program in rats fed with standard chow induced right atria subsensitivity to noradrenaline and supersensitivity to isoprenaline. Atria isolated from rats fed with high fat-CHO diet showed subsensitivity to noradrenaline and no alterations on the sensitivity to isoprenaline. Association of high fat-CHO diet and this physical exercise program prevented the alterations induced by the diet or the exercise values were: 7.44±0.09 (NS); 6.65±0.17* (HS); 6.52±0.25* (NT), 7.15±0.04**(HT) and ISO pDprograms alone. The NA pD2 # 2 values were: 8.37±0.12 (NS); 8.52±0.10 (HS); 8.94±0.09* (NT), 8.55±0.08 (HT). Significant differences (p<0.05 ANOVA plus Tukey test) were indicated as follows: *compared to the NS group, **compared to the NT group, #compared to the HS group. The uptake inhibition did not alter the responses obtained. This exercise program induced alterations in the right atria response similar to those induced by foot shock stress, it means, decrease on ß1-adrenoceptor mediated (NA) response and increase on ß2-adrenoceptor mediated (ISO) response. High fat-CHO diet offered during four weeks induced subsensitivity to noradrenaline, this response is probably due to alterations on the lipid membrane content and high adrenergic activity reported in this animal model. The absence of effects on atria response observed in rats submitted to high fat-CHO diet and exercise program needs to be investigated. / Mestrado / Fisiologia / Mestre em Biologia Funcional e Molecular

Au cours de l’administration simultanée de plusieurs médicaments injectables, sont utilisées une ligne principale de perfusion et une ou plusieurs ligne(s) en dérivation. La ligne principale est directement reliée au cathéter et permet l’administration des solutions utilisées pour l’apport hydro-électrolytique ou de maintenir une voie d’abord veineuse perméable. Les autres thérapeutiques vont être perfusées en dérivation de cette ligne principale. La prise en charge des patients relevant de soins intensifs et de réanimation nécessite une polymédication. Les traitements d’urgence impliquent majoritairement des médicaments à marge thérapeutique étroite. Lors de l’administration de médicaments en solutions concentrées, de faibles perturbations du débit d’administration peuvent engendrer de fortes perturbations cliniques et notamment, pour les médicaments vasoactifs, créer une instabilité hémodynamique. C’est pourquoi il est important d’étudier la problématique de la perfusion simultanée, de déterminer l’impact sur le débit massique des lignes de perfusion et la technique optimale des changements de seringues pour prévenir les variations intempestives du débit de perfusion. Ce mémoire présente un travail de développement et d’évaluation d’une démarche d’optimisation d’un système de perfusion complexe. Il consiste à étudier au travers d’évaluations in vitro et d’études cliniques la conception d’une ligne de perfusion en évaluant notamment un dispositif médical innovant afin de proposer, in fine, une solution applicable dans un service de soins intensifs. La première partie consiste à présenter l’ensemble des dispositifs médicaux de perfusion utilisés dans un département d’anesthésie-réanimation. La seconde partie s’intéresse à l’administration d’un médicament couramment perfusé sur la voie proximale: la noradrénaline. Les études in vitro, corroborées par des données cliniques, ont permis de montrer la supériorité de l’administration de noradrénaline à 0,5 mg/mL perfusée en Y avec une solution saline isotonique à débit fixe de 5 mL/h. Cette multiperfusion fait intervenir l’utilisation d’un prolongateur trois voies à faible volume résiduel, permettant d’optimiser les conditions de relais de seringues, connues comme étant à l’origine d’instabilités hémodynamiques chez les patients traités par catécholamines. Un programme hospitalier de recherche clinique interrégional est déposé dans le but d’établir des recommandations de perfusion des catécholamines.La troisième partie aborde l’administration des médicaments sur voie distale en sélectionnant l’insuline comme marqueur-médicament. Les résultats de cette étude clinique prospective randomisée contrôlée ont montré que l’utilisation d’un dispositif médical innovant, le dispositif Edelvaiss-Multiline 8 (Doran International) caractérisé par un tube multilumières à faible volume résiduel qui permet de dédier une voie à une seule thérapeutique, permettait de réduire significativement le temps passé en hypoglycémie pour 1000 heures de perfusion d’insuline au cours de perfusion continue d’insuline en soins intensifs périopératoires.Enfin, dans une dernière partie, les critères clés d’un montage optimisé de multiperfusion sont élaborés et sont mis en application dans un département d’anesthésie-réanimation dans le but d’optimiser et uniformiser la ligne de perfusion des patients. Ce travail a permis de valider les caractéristiques clés de la ligne de perfusion définis dans de précédentes études non cliniques : la nature du matériau des dispositifs médicaux utilisés, l’utilisation de valves appropriées, la minimisation des volumes internes des tubulures de perfusion, l’utilisation de systèmes de perfusion automatisés permettant de contrôler au mieux le débit d’administration des médicaments. / For the simultaneous administration of injectable drugs, the infusion line includes a main line with one or several derivative lines. The main line, which is directly connected to the catheter, is dedicated to hydration infusion or to maintain a permeable vein. Other medications will be added on the derivative lines.Intensive care unit patients frequently require lots of medications in the same time. Most of emergency drugs are substances with narrow therapeutic range. When concentrated solutions are employed, tiny mass flow rate disturbances can provoke clinical damages, such as haemodynamic instability. So, several parameters have been studied on simultaneous infusions: mass flow rate and syringes changeovers.The purpose of this work was to develop and optimize complex infusion line systems. An innovative infusion medical device has been evaluated in clinical trials and in vitro studies. The final objective was to design an optimized infusion line, which could be applied to ICUs.The whole medical devices used in ICUs was first listed. Then, noradrenaline has been used as the reference drug to study central venous catheter proximal line. A 0.5 mg/mL noradrenaline solution Y-infused with a saline (5mL/h) has been shown by clinical and in vitro data to be the best solution. Nevertheless, this conclusion was valid only with the use of a very low dead-space volume Y-extension set. Thanks to this device, syringe changeovers optimization is possible.The central venous catheter distal line has been studied in a second time through an open randomized controlled prospective clinical trial. Primary endpoint of the study was the impact of two different insulin infusion lines (Edelvaiss-Multiline 8, Doran International versus standard line) on glycaemic variability. Doran’s innovative device consists of an exten¬sion set with eight accesses connected to nine separated lumens in a single tube. This allows to dedicate an isolated way for insulin. With its use, a significant decrease of hypoglycaemia occurring in 1000h of infusion period was clinically demonstrated. Finally, all the data were synthetized to optimize an ICU multi-infusion line. The one, which has been designed for surgery and intensive care units, was tested on patients.To conclude, items responsible for mass flow rate disturbances have been identified: medical devices material, addition of appropriated valves, internal volume line minimization and use of automated infusion systems (as pumps). The ideal infusion line has to take into account all these parameters.

Avaliar o efeito in vitro da adrenalina, noradrenalina e cortisol sobre a formação do biofilme, produção de ácido e expressão de fatores de virulência por S. mutans. Método: Para a formação do biofilme foi realizada uma monocultura de S. mutans (UA159) em discos de hidroxiapatita (HA) associados à adrenalina, noradrenalina e cortisol por cinco dias. Após esse período, foi realizada a contagem bacteriana (ufc/ml) para cada grupo experimental. A avaliação da queda do pH (produção de ácidos) foi realizada a cada 12 horas até o final do experimento (cinco dias). Para a avaliação da expressão de fatores de virulência, RNA total de biofilme maduro (cinco dias) foi extraído, e a análise da expressão de genes relacionados com a virulência do S. mutans (gtfB, gtfC, gtfD, brpA e ldh) foi realizada através de RT-qPCR. Resultados: As catecolaminas (adrenalina e noradrenalina) e também o cortisol aumentaram significativamente a formação de colônias de S. mutans em relação ao grupo controle. Uma queda no valor do pH foi observada nas 12 primeiras horas em todos os grupos. Após este período, os valores se mantiveram praticamente estáveis até o final do experimento (120h), não havendo diferença estatística entre o grupo controle e os grupos testes. Em relação ao efeito das catecolaminas/cortisol sobre a expressão de genes de virulência do S. mutans, não se observou diferença estatística significativa entre os diferentes grupos. Conclusões: Os achados do presente estudo demonstraram que a adrenalina, noradrenalina e cortisol aumentam a formação do biofilme (in vitro), sem alterar o pH (produção de ácidos) e expressão dos genes de virulência avaliados. / To evaluate the in vitro effect of adrenaline, noradrenaline and cortisol on biofilm formation, acid production and expression of virulence factors by S. mutans. Methods: Biofilm formation was performed by a monoculture of S. mutans (UA159) in discs of hydroxyapatite (HA) associated with adrenaline, noradrenaline and cortisol for five days. Further, bacterial count was performed (cfu / ml) for each experimental group. Evaluation of the reduction in pH (acid production) was conducted then every 12 hours until the end of treatment (five days). For evaluation of the virulence factors of mature biofilm, total RNA (five days) was extracted and the analysis of expression of genes related to the virulence of S. mutans (gtfB, gtfC, gtfD, brpA and ldh) was performed by RT-qPCR. Results: Catecholamines (adrenaline and noradrenaline) and cortisol tested in this study significantly increased the formation of colonies of S. mutans in the control group. In all groups (control, adrenaline, noradrenaline and cortisol) the highest reduction in pH was observed during the first 15 minutes, with no statistical difference between the control and test group. PH measurements were performed until the 5th day of the experiment at intervals of 12h. Continuous reduction was observed during the first 12 hours in all groups after this period, the values remained almost stable until the end of the experiment (120h). There was no statistical difference between the control and test groups. Regarding the effect of catecholamine / cortisol on the expression of virulence genes of the S. mutans, there was no statistically significant difference among the groups and the control group. Conclusions: The findings of this study demonstrated that adrenaline, norepinephrine and cortisol increase the formation of biofilm (in vitro), without changing the pH (initial and in biofilm) and expression of genes evaluated.

The establishment of physiological norms for psychologically hardy vs. non-hardy individuals was attempted by examination of levels of salivary cortisol and urinary norepinephrine before and after a mid-term examination stressor. Normative data was collected on the reported frequency of stressors and their severity one week prior to the examination, and self-reported ratings of stress immediately prior to the examination. Performance on the examination as a function of hardiness was explored. Associations between demographic variables and psychological hardiness were also studied. Results from this study were inconclusive in establishing physiological norms for psychologically hardy individuals. Associations were found between: 1) hardiness and frequency of stressors; 2) hardiness and age; and 3) self-reported ratings of stress and anxiety as measured by the State-Trait Anxiety Inventory (STAI).

The locus coeruleus (LC) projects ubiquitously to the cortex, and noradrenaline (NA) exerts powerful neuromodulatory control on cortical excitation and inhibition. Previous work has shown that NA plays an important role in motor processes, and further posits that dysregulation in NA function could be one of the culprits of motor-related deficits in many neurodevelopmental disorders, including Autism Spectrum Disorder. In order to characterize the change in NA levels during motor learning in awake and behaving mice, I employed a newly developed optical NA sensor, combined with in vivo two-photon imaging, to visualize spatiotemporal activation patterns of NA in the motor cortex. This experimental approach allows us to track and chronically image the same region of the motor cortex over multiple days, thus permitting the characterization of NA activity throughout the entirety of the motor learning process. I found that NA levels increase significantly during the initial phase of learning, which coincides with the structural and functional plastic changes that have been previously reported in the motor cortex during early stages of motor learning. The NA activity returns to baseline levels as the mice develop their movement strategy; however, the regions of NA release become more spatially clustered during the learning process. The results reported in this thesis provide a novel glimpse into the dynamics of NA activity in the motor cortex during motor learning, and it will provide new direction for the development of therapeutic strategies and diagnostic criteria for motor-related dysfunction in neurodevelopmental diseases.

Bugula neritina is a sessile marine bryozoan with a pelagic larval stage. Larvae frequently settle on boat hulls, facilitating the introduction of B. neritina to bays and estuaries worldwide. Adrenergic agonists, such as the vertebrate hormone noradrenaline, inhibit larval settlement in a variety of marine invertebrate species, including B. neritina. Light also inhibits B. neritina larval settlement, yet the underlying mechanisms by which light and adrenergic compounds exert their effects on larvae are not well understood. Octopamine is considered the invertebrate analog of noradrenaline, and may be an endogenous hormone involved in larval settlement pathways. I observed the effects of the adrenergic agonist noradrenaline and the adrenergic antagonist phentolamine on larval settlement, and found that high concentrations of noradrenaline increased larval mortality, inhibited larval attachment, and increased larval swimming behavior. High concentrations of phentolamine also increased larval mortality, but increased larval attachment and decreased larval swimming behavior. I used fluorescent labeling and microscopy to localize sensory system components, and found that larvae possess adrenergic-like receptors, as well as tyrosine hydroxylase-like and octopamine-like immunoreactivity. I also exposed larvae to phentolamine in both dark and light conditions, and found that light significantly inhibited larval attachment, but phentolamine blocked those inhibitory effects. These results suggest that B. neritina larvae possess adrenergic-like receptors, which serve as the binding sites for noradrenaline and phentolamine. These are likely octopamine receptors, and octopamine may be one endogenous compound involved in controlling larval phototaxis and settlement behavior. Light may increase octopamine production, thereby stimulating cilial activity, extending swimming behavior, and preventing larvae from attaching to a substrate. This research sheds light on previously unknown sensory mechanisms in B. neritina larvae, and may aid in the development of new biofouling control strategies.

No description available.

Yes / Here we describe for the first time the distinctive pharmacological profile for IRL752, a new phenyl-pyrrolidine derivative with regio-selective CNS transmission-enhancing properties. IRL752 (3.7-150 μmol/kg, s.c.) was characterised through extensive in vivo studies, using behavioural, tissue neurochemical and gene expression, as well as microdialysis methods. Behaviourally, the compound normalised tetrabenazine-induced hypoactivity, while unable to stimulate basal locomotion in normal animals or to either accentuate or reverse hyperactivity induced by amphetamine or MK-801. IRL752 induced but minor changes in monoaminergic tissue neurochemistry across NA- and DA-dominated brain regions. The expression of neuronal activity-, plasticity-, and cognition-related IEGs (immediate early genes) however increased by 1.5- to 2-fold. Furthermore, IRL752 dose-dependently enhanced cortical catecholamine dialysate output to 600-750% above baseline, while striatal DA remained unaltered and NA rose to ~250%; cortical and hippocampal dialysate ACh increased to ~250% and 190% above corresponding baseline, respectively. In line with this cortically preferential transmission-promoting action, the drug was also pro-cognitive in the novel object recognition and reversal learning tests. In vitro neurotarget affinity and functional data, coupled to drug exposure support the hypothesis that 5‑HT7 receptor and α2(C)-adrenoceptor antagonism are key contributors to the in vivo efficacy and original profile of IRL752. The cortical-preferring facilitatory impact on catecholamine (and ACh) neurotransmission, along with effects on IEG expression and cognition-enhancing features, are in line with the potential clinical usefulness of IRL752 in conditions where these aspects may be dysregulated, such as in axial motor and cognitive deficits in Parkinson's Disease.