O núcleo do trato solitário é uma área integrativa do sistema nervoso central (CNS) envolvida no controle autonômico e respiratório. Estudos da literatura sugerem que o óxido nítrico (NO) exerce um importante papel na modulação dos reflexos cardiovasculares e ventilatórios no NTS. Além disso, evidências da literatura indicam uma possível interação entre o NO e o ATP no SNC. Fundamentados nessas evidências, no presente estudo, avaliamos a possível participação do NO na modulação dos parâmetros cardiorespiratórios basais e no processamento das respostas cardiovasculares e respiratórias à ativação do quimiorreflexo no NTS caudal de ratos não-anestesiados. Além disso, o possível papel do NO produzido pela óxido nítrico sintase neuronal (nNOS) nas respostas cardiovasculares e respiratórias à microinjeção unilateral de ATP no NTS caudal também foi avaliado. Para tanto, os animais foram submetidos ao implante de cânulas guia em direção ao NTS caudal e à canulação da artéria e veia femoral. Os parâmetros ventilatórios foram avaliados pelo método de pletismografia de corpo inteiro. A análise dos resultados monstrou que as microinjeções bilaterais do L-NAME, um inibidor nãoseletivo da NOS, no NTS caudal, promoveram um aumento significativo na pressão arterial basal dos animais, sugerindo um papel modulatório do NO sobre os neurônios envolvidos com as vias neurais do barorreflexo. No entanto, as microinjeções bilaterais do N-PLA, um inibidor seletivo da nNOS, no NTS caudal, não promoveram alterações significativas na pressão arterial basal, sugerindo que a produção do NO envolvido no controle autonômico basal no NTS caudal não é dependente da atividade da nNOS. Com relação às respostas do quimiorreflexo, as microinjeções bilaterais do L-NAME ou do N-PLA, no NTS caudal de ratos nãoanestesiados, promoveram um atenuação significativa no aumento da freqüência respiratória (fR) à ativação do quimiorreflexo, sugerindo a participação do NO Resumo xii produzido pela nNOS na modulação do componente respiratório do quimiorreflexo no NTS caudal. No entanto, as respostas pressora e bradicárdicas decorrentes da ativação do quimiorreflexo não foram alteradas pelas microinjeções bilaterais do LNAME ou N-PLA no NTS caudal, sugerindo que o NO não está envolvido na modulação das respostas cardiovasculares decorrentes da ativação deste reflexo. No que diz respeito às respostas decorrentes da microinjeção de ATP no NTS caudal, a análise dos resultados demonstrou que as respostas de aumento na pressão arterial, fR e ventilação minuto produzidas pela microinjeção unilateral de ATP no NTS caudal de ratos não-anestesiados foram significativamente atenuadas após a microinjeção do N-PLA no mesmo sítio, sugerindo a participação do NO produzido pela nNOS na modulação de tais respostas. Neste contexto, os achados do presente trabalho sugerem que no NTS caudal: 1) o NO, provavelmente produzido pela NOS endotelial, exerce um importante papel modulatório nas vias neurais do barorreflexo; 2) a neurotransmissão do aumento da fR decorrente da ativação do quimiorreflexo envolve a formação de NO produzido pela nNOS; 3) a neurotransmissão das respostas cardiovasculares decorrentes da ativação do quimiorreflexo não envolve a formação de NO; 4) a neurotransmissão das respostas cardiovascualres e respiratórias decorrentes da microinjeção unilateral de ATP envolve a formação de NO produzido pela nNOS. / The nucleus tractus solitarius (NTS) is an integrative area in the central nervous system (CNS) involved with the ventilatory and autonomic control. Several studies suggest that nitric oxide (NO) in the NTS plays an important role in the modulation of the cardiovascular and ventilatory reflexes. In addition, there is evidence indicating a possible interaction of NO and ATP in the CNS. Considering these findings, in the present study, we evaluated the possible role of NO on the modulation of the basal cardiorespiratory parameters as well as on the processing of the cardiovascular and ventilatory responses elicited by chemoreflex activation in the caudal NTS of awake rats. In addition, the possible role of NO produced by neuronal nitric oxide sintase (nNOS) on the cardiovascular and respiratory responses produced by unilateral microinjection of ATP into the caudal NTS was also evaluated. For this purpose, rats received bilateral guide cannulae in direction of the caudal NTS and femoral artery and vein were cannulated. The ventilatory measurements were obtained by whole-body pletismograph method. Our data showed that bilateral microinjections of L-NAME, a non-selective NOS inhibitor, into the caudal NTS, produced a significant increase in basaline mean arterial pressure, suggesting a modulatory role of NO in the neural pathways of the baroreflex. However, bilateral microinjections of N-PLA, a selective nNOS inhibitor, into the caudal NTS, produced no significant changes in the baseline mean arterial pressure, suggesting that NO produced by nNOS is not involved in the basal autonomic control in the caudal NTS. With respect to chemoreflex responses, bilateral microinjections of L-NAME or NPLA, into the caudal, produced a significant attenuation in the increase in respiratory frequency (fR) produced by chemoreflex activation, suggesting that NO produced by nNOS is involved in the modulation of the respiratory component of the chemoreflex. However, the pressor and bradicardic responses elicited by chemoreflex actiovation Abstract xv were not affected by microinjections of L-NAME or N-PLA, suggesting that NO is not involved in the modulation of the cardiovascular responses. With respect to ATP microinjection responses, the data showed that unilateral microinjection of ATP into the caudal NTS produced increase in arterial pressure, fR and minute ventilation, which were significantly attenuated by N-PLA, suggesting that NO produced by nNOS is involved in the modulation of the cardiovascular and ventilatory responses to ATP microinjection into the caudal NTS. In conclusion, the data of present study indicate that in the caudal NTS: 1) NO, produced probably by endothelial NOS, plays an important modulatory role on the neural pathways of the baroreflex; 2) the neurotransmission of the increase in respiratory frequency to chemoreflex activation involve NO production by nNOS. 3) NO is not involved in modulation of the autonomic components of chemoreflex; 4) the cardiovascular and ventillatory responses produced by ATP micronjection are, at least in part, mediated by NO produced by nNOS.

alpha-melanocyte stimulating hormone (alpha-MSH) is an important regulator of food intake, metabolic rate, and inflammation. Recently, alpha-MSH was shown to influence sympathetic activity and blood pressure regulation. In the present study, we investigated the cardiovascular effects of alpha-MSH in the nucleus tractus solitarii (NTS) of spontaneously hypertensive rats (SHR). Because nitric oxide (NO) is well-known to involve in central cardiovascular regulation, we elucidated the role of NO in the cardiovascular responses induced by alpha-MSH. In urethane-anesthetized SHR, unilateral microinjection of alpha-MSH (0.3-300 pmol) into the NTS produced dose-responsive depressor and bradycardic effects. The cardiovascular effects of alpha-MSH were abrogated by the antagonist of melanocortin receptor (MC3/4-R), SHU9119. Pretreatment with precursor of nitric oxide, L-arginine, enhanced the duration of alpha-MSH-mediated hypotensive effects, whereas prior application of L-NAME, a universal inhibitor of nitric oxide synthase (NOS), significantly attenuated the effects of alpha-MSH. Prior injection with inhibitor of inducible NOS, aminoguanidine, but not inhibitor of neuronal NOS, 7-nitroindazole, attenuated the hypotensive effect of alpha-MSH. In summary, these results indicated alpha-MSH induced depressor and bradycardic effects in the NTS of SHR. Besides, the hypotensive mechanism of alpha-MSH was mediated via MC4-R and involved with iNOS activation in the NTS of SHR.

No description available.

Intensive Forschung hat in den vergangenen Jahrzehnten zu einer sehr detaillierten Charakterisierung des Geschmackssystems der Säugetiere geführt. Dennoch sind mit den bislang eingesetzten Methoden wichtige Fragestellungen unbeantwortet geblieben. Eine dieser Fragen gilt der Unterscheidung von Bitterstoffen. Die Zahl der Substanzen, die für den Menschen bitter schmecken und in Tieren angeborenes Aversionsverhalten auslösen, geht in die Tausende. Diese Substanzen sind sowohl von der chemischen Struktur als auch von ihrer Wirkung auf den Organismus sehr verschieden. Während viele Bitterstoffe potente Gifte darstellen, sind andere in den Mengen, die mit der Nahrung aufgenommen werden, harmlos oder haben sogar positive Effekte auf den Körper. Zwischen diesen Gruppen unterscheiden zu können, wäre für ein Tier von Vorteil. Ein solcher Mechanismus ist jedoch bei Säugetieren nicht bekannt. Das Ziel dieser Arbeit war die Untersuchung der Verarbeitung von Geschmacksinformation in der ersten Station der Geschmacksbahn im Mausgehirn, dem Nucleus tractus solitarii (NTS), mit besonderem Augenmerk auf der Frage nach der Diskriminierung verschiedener Bitterstoffe. Zu diesem Zweck wurde eine neue Untersuchungsmethode für das Geschmackssystem etabliert, die die Nachteile bereits verfügbarer Methoden umgeht und ihre Vorteile kombiniert. Die Arc-catFISH-Methode (cellular compartment analysis of temporal activity by fluorescent in situ hybridization), die die Charakterisierung der Antwort großer Neuronengruppen auf zwei Stimuli erlaubt, wurde zur Untersuchung geschmacksverarbeitender Zellen im NTS angewandt. Im Zuge dieses Projekts wurde erstmals eine stimulusinduzierte Arc-Expression im NTS gezeigt. Die ersten Ergebnisse offenbarten, dass die Arc-Expression im NTS spezifisch nach Stimulation mit Bitterstoffen auftritt und sich die Arc exprimierenden Neurone vornehmlich im gustatorischen Teil des NTS befinden. Dies weist darauf hin, dass Arc-Expression ein Marker für bitterverarbeitende gustatorische Neurone im NTS ist. Nach zweimaliger Stimulation mit Bittersubstanzen konnten überlappende, aber verschiedene Populationen von Neuronen beobachtet werden, die unterschiedlich auf die drei verwendeten Bittersubstanzen Cycloheximid, Chininhydrochlorid und Cucurbitacin I reagierten. Diese Neurone sind vermutlich an der Steuerung von Abwehrreflexen beteiligt und könnten so die Grundlage für divergentes Verhalten gegenüber verschiedenen Bitterstoffen bilden. / Intense research in the past decades has led to a detailed understanding of the mammalian taste system. Some important issues, however, have remained unanswered with the established methods that have been applied so far. One of these questions is whether different bitter substances can be distinguished. There are thousands of compounds which taste bitter to humans and elicit innate aversive behavior in animals. Moreover, these bitter substances are very heterogeneous regarding their structure as well as their effect on the organism. While many bitter tastants are potent poisons, others are harmless or even have beneficial effects in the amounts that are typically ingested. The ability to discriminate between those groups of bitter tastants could be an evolutionary advantage. Such a mechanism, however, is not known for mammals. The aim of this thesis was to study the processing of taste information in the first station of gustatory processing in the mouse brain, the nucleus of the solitary tract (NTS). Of particular interest was the question concerning discrimination of bitter tastants. To this end a new method was established for the taste system combining the advantages of methods used before while circumventing their disadvantages. The Arc catFISH method (cellular compartment analysis of temporal activity by fluorescent in situ hybridization), which allows the characterization of responses of large neuron populations to two stimuli, was used to analyze taste-processing cells in the NTS. In the course of this project a stimulus-induced Arc expression in the NTS was shown for the first time. The results demonstrated that Arc expression in the NTS appears specifically after stimulation with bitter tastants and that the Arc expressing neurons are located primarily in the gustatory part of the NTS. This indicates that Arc expression is a marker for bitter-processing gustatory neurons in the NTS. Upon stimulating twice with bitter compounds, distinct, yet overlapping neuron populations were identified, that reacted differently to the three bitter substances cycloheximide, quinine hydrochloride, and cucurbitacin I. Presumably these neurons are involved in the regulation of aversive reflexes and could form a basis for divergent behavior towards different bitter substances.