Esponjas (filo Porifera) são consideradas representantes atuais dos primeiros metazoários, com origens evolutivas próximas da transição de uma organização unicelular para aquela multicelular. Toda sua fisiologia é baseada em células especializadas, sem órgãos ou tecidos verdadeiros como encontrados em outros animais. No entanto, a presença de um sistema fortemente integrado pode ser constatada em sua habilidade de contrair o corpo de maneira rítmica e coordenada, ainda que a dinâmica e o controle desse processo sejam pouco conhecidos. Neste trabalho, análises de imagens em time-lapse sob um ciclo claro-escuro (12:12h) e sob luz constante mostram que as espécies Hymeniacidon heliophila e Tethya maza exibem padrões distintos de contrações corpóreas rítmicas, com variações circadianas em suas amplitudes. Diferenças observadas nos perfis das curvas das duas espécies podem ser devidas a diferenças em seus habitats de origem e/ou a diferenças em suas estruturas corpóreas. A manutenção desses ritmos sob luz constante indica a presença de um relógio endógeno em ambos os animais. Entre as vias envolvidas na sinalização desses processos a mais ubíqua é a da melatonina, um hormônio cuja presença tem sido demostrada em muitos filos animais, dos quais o mais basal é o Cnidaria. Considerando a ampla ação da melatonina como um sinalizador circadiano e a ausência de um sistema nervoso em esponjas, não seria surpreendente constatar um papel da molécula na coordenação dos movimentos desses animais. Ademais, o papel da molécula nas respostas imunes também já foi demonstrado ser de ampla distribuição. Nossas tentativas de revelar uma ação da molécula na reagregação celular de H. heliophila via receptores do tipo MT1 e MT2 não foram conclusivas. Alguns fatores que podem ter influenciado nas respostas divergentes da molécula são o estado imunitário e a presença de contaminantes no ambiente de origem do animal. Ainda, outros mecanismos de ação e/ou processos candidatos à ação da melatonina não devem ser descartados. Ainda que preliminares, nossos resultados são o primeiro relato da presença da melatonina no filo mais basal dentre os Metazoa / Sponges (phylum Porifera) are considered current representatives of the first metazoans, with evolutionary origins close to the transition from a unicellular to a multicellular organization. All of its physiology is based on specialized cells, without organs or real tissues as found in other animals. However, the presence of a strongly integrated system can be verified in its ability to contract the body in a rhythmic and coordinated way, althouth the dynamic and the control of this process are little known. In this work, time-lapse image analysis under a light-dark cycle (12:12h) and constant light shows that the species Hymeniacidon heliophila and Tethya maza display different patterns of body contractions, with circadian variations in their amplitudes. These dissimilar contraction profiles of the two species can be due to their original habitats and/or in their body structures. The maintenance of these rhythms under constant light indicates the presence of an endogenous clock in both animals. Among the pathways that participate in this process the most ubiquitous is that of melatonin, a hormone whose presence has been demonstrated in several animals phyla, of which the most basal until now is Cnidaria. Considering the melatonin action as a circadian signal and the absence of a nervous system in sponges, it would not be surprising that this molecule participates in coordination in these animals. In addition, the melatonin role in the immune responses has also been shown to be widely distributed. Our attempts to reveal a molecule action in the H. heliophila cellular reaggregation via MT1 and MT2 receptors were inconclusive. Some of the factors that may have influenced the molecule divergent responses are the immunity condition and the presence of contaminants in the original environment of the animals. Still, other mechanisms and/or processes that are candidates for melatonin action should not be discarded. Although preliminary, our results are the first report of the presence of melatonin in the most basal phylum among the Metazoa

Esponjas (filo Porifera) são consideradas representantes atuais dos primeiros metazoários, com origens evolutivas próximas da transição de uma organização unicelular para aquela multicelular. Toda sua fisiologia é baseada em células especializadas, sem órgãos ou tecidos verdadeiros como encontrados em outros animais. No entanto, a presença de um sistema fortemente integrado pode ser constatada em sua habilidade de contrair o corpo de maneira rítmica e coordenada, ainda que a dinâmica e o controle desse processo sejam pouco conhecidos. Neste trabalho, análises de imagens em time-lapse sob um ciclo claro-escuro (12:12h) e sob luz constante mostram que as espécies Hymeniacidon heliophila e Tethya maza exibem padrões distintos de contrações corpóreas rítmicas, com variações circadianas em suas amplitudes. Diferenças observadas nos perfis das curvas das duas espécies podem ser devidas a diferenças em seus habitats de origem e/ou a diferenças em suas estruturas corpóreas. A manutenção desses ritmos sob luz constante indica a presença de um relógio endógeno em ambos os animais. Entre as vias envolvidas na sinalização desses processos a mais ubíqua é a da melatonina, um hormônio cuja presença tem sido demostrada em muitos filos animais, dos quais o mais basal é o Cnidaria. Considerando a ampla ação da melatonina como um sinalizador circadiano e a ausência de um sistema nervoso em esponjas, não seria surpreendente constatar um papel da molécula na coordenação dos movimentos desses animais. Ademais, o papel da molécula nas respostas imunes também já foi demonstrado ser de ampla distribuição. Nossas tentativas de revelar uma ação da molécula na reagregação celular de H. heliophila via receptores do tipo MT1 e MT2 não foram conclusivas. Alguns fatores que podem ter influenciado nas respostas divergentes da molécula são o estado imunitário e a presença de contaminantes no ambiente de origem do animal. Ainda, outros mecanismos de ação e/ou processos candidatos à ação da melatonina não devem ser descartados. Ainda que preliminares, nossos resultados são o primeiro relato da presença da melatonina no filo mais basal dentre os Metazoa / Sponges (phylum Porifera) are considered current representatives of the first metazoans, with evolutionary origins close to the transition from a unicellular to a multicellular organization. All of its physiology is based on specialized cells, without organs or real tissues as found in other animals. However, the presence of a strongly integrated system can be verified in its ability to contract the body in a rhythmic and coordinated way, althouth the dynamic and the control of this process are little known. In this work, time-lapse image analysis under a light-dark cycle (12:12h) and constant light shows that the species Hymeniacidon heliophila and Tethya maza display different patterns of body contractions, with circadian variations in their amplitudes. These dissimilar contraction profiles of the two species can be due to their original habitats and/or in their body structures. The maintenance of these rhythms under constant light indicates the presence of an endogenous clock in both animals. Among the pathways that participate in this process the most ubiquitous is that of melatonin, a hormone whose presence has been demonstrated in several animals phyla, of which the most basal until now is Cnidaria. Considering the melatonin action as a circadian signal and the absence of a nervous system in sponges, it would not be surprising that this molecule participates in coordination in these animals. In addition, the melatonin role in the immune responses has also been shown to be widely distributed. Our attempts to reveal a molecule action in the H. heliophila cellular reaggregation via MT1 and MT2 receptors were inconclusive. Some of the factors that may have influenced the molecule divergent responses are the immunity condition and the presence of contaminants in the original environment of the animals. Still, other mechanisms and/or processes that are candidates for melatonin action should not be discarded. Although preliminary, our results are the first report of the presence of melatonin in the most basal phylum among the Metazoa

<p>The duodenal epithelium secretes bicarbonate ions and this is regarded as the primary defence mechanism against the acid discharged from the stomach. For an efficient protection, the duodenum must also function as a sensory organ identifying luminal factors. Enteroendocrine cells are well-established intestinal “taste” cells that express signaling peptides such as orexins and guanylins. Luminal factors affect the release of these peptides, which may modulate the activity of nearby epithelial and neural cells.</p><p>The present thesis considers the effects of orexins and guanylins on duodenal bicarbonate secretion. The duodenal secretory response to the peptides was examined in anaesthetised rats <i>in situ</i> and the effects of orexin-A on intracellular calcium signaling by human as well as rat duodenal enterocytes were studied <i>in vitro</i>.</p><p>Orexin-A, guanylin and uroguanylin were all stimulants of bicarbonate secretion. The stimulatory effect of orexin-A was inhibited by the OX₁-receptor selective antagonist SB-334867. The muscarinic antagonist atropine on the other hand, did not affect the orexin-A-induced secretion, excluding involvement of muscarinic receptors. Orexin-A induced calcium signaling in isolated duodenocytes suggesting a direct effect at these cells. Interestingly, orexin-induced secretion and calcium signaling as well as mucosal orexin-receptor mRNA and OX₁-receptor protein levels were all substantially downregulated in overnight fasted rats compared with animals with continuous access to food. Further, secretion induced by Orexin-A was shown to be dependent on an extended period of glucose priming.</p><p>The uroguanylin-induced bicarbonate secretion was reduced by atropine suggesting involvement of muscarinic receptors. The melatonin receptor antagonist luzindole attenuated the secretory response to intra-arterially administered guanylins but had no effect on secretion when the guanylins were given luminally. </p><p>In conclusion, the results suggest that orexin-A as well as guanylins may participate in the regulation of duodenal bicarbonate secretion. Further, the duodenal orexin system is dependent on the feeding status of the animals.</p>

The duodenal epithelium secretes bicarbonate ions and this is regarded as the primary defence mechanism against the acid discharged from the stomach. For an efficient protection, the duodenum must also function as a sensory organ identifying luminal factors. Enteroendocrine cells are well-established intestinal “taste” cells that express signaling peptides such as orexins and guanylins. Luminal factors affect the release of these peptides, which may modulate the activity of nearby epithelial and neural cells. The present thesis considers the effects of orexins and guanylins on duodenal bicarbonate secretion. The duodenal secretory response to the peptides was examined in anaesthetised rats in situ and the effects of orexin-A on intracellular calcium signaling by human as well as rat duodenal enterocytes were studied in vitro. Orexin-A, guanylin and uroguanylin were all stimulants of bicarbonate secretion. The stimulatory effect of orexin-A was inhibited by the OX1-receptor selective antagonist SB-334867. The muscarinic antagonist atropine on the other hand, did not affect the orexin-A-induced secretion, excluding involvement of muscarinic receptors. Orexin-A induced calcium signaling in isolated duodenocytes suggesting a direct effect at these cells. Interestingly, orexin-induced secretion and calcium signaling as well as mucosal orexin-receptor mRNA and OX1-receptor protein levels were all substantially downregulated in overnight fasted rats compared with animals with continuous access to food. Further, secretion induced by Orexin-A was shown to be dependent on an extended period of glucose priming. The uroguanylin-induced bicarbonate secretion was reduced by atropine suggesting involvement of muscarinic receptors. The melatonin receptor antagonist luzindole attenuated the secretory response to intra-arterially administered guanylins but had no effect on secretion when the guanylins were given luminally. In conclusion, the results suggest that orexin-A as well as guanylins may participate in the regulation of duodenal bicarbonate secretion. Further, the duodenal orexin system is dependent on the feeding status of the animals.