Les canaux activés par l’hyperpolarisation et sensibles aux nucléotides cycliques (hy- perpolarization cyclic nucleotide-gated channel; HCN) ont un rôle dans la régulation du rythme cardiaque ainsi que dans la transmission des influx nerveux. De nombreuses pathologies sont causées par un disfonctionnement de ces canaux. A ce jour, Ivabradine représente la seule molécule utilisée comme médicament, mais malgré sa spécificité en- vers les canaux HCNs, elle n’est pas sélective entre les isoformes de la famille HCN. Dans le but d’identifier une molécule plus adéquate pour le traitement qu’Ivabradine, nous avons analysé l’interaction entre les canaux HCNs et 9 bloqueurs connus garce à l’arrimage moléculaire. Cette analyse nous a permis d’identifier les résidus nécessaires pour la liaison des ligands. On observe aussi qu’Ivabradine ne forme aucune liaison so- lide avec les canaux HCNs mais ne fait que bloquer le passage par sa présence tout comme ses dérivées Zatebradine et Cilobradine. Les ligands de plus petites tailles quant à eux, se logent dans une cavité hydrophobe et forme des liaisons stable avec les pro- téines. Nos résultats semblent suggérer que le blocage par Ivabradine est plus efficace, mais que les liaisons stables des petits ligands possèdent un potentiel plus grand vers une meilleure affinité. Par contre, les interactions observées suggèrent que la spécificité envers les isoformes proviendrait des cinétiques des canaux et des dépendances d’états des ligands plutôt que des interactions identifiées. Pour finir, l’arrimage des ligands sur la conformation fermé du canal HCN1 suggère qu’il existerait une conformation fermée- liée non connue puisqu’aucun ligand n’a pu accéder au pore. / HCN channels have a role in regulating heart rate as well as in the transmission of nerve impulses. Many pathologies are caused by a malfunction of these channels. To date, Ivabradine is the only molecule used as a drug, but despite its specificity for HCN chan- nels, it is not selective between the isoforms of the HCN family. In order to identify a molecule more suitable for the treatment than ivabradine, we analyzed the interaction between HCN family and 9 known blockers using docking. This analysis allowed us to identify the necessary residuals for ligand binding. It is also observed that ivabradine forms no solid bond with the HCN channels but only blocks the passage by its pres- ence, the same was observed for its derivatives Zatebradine and Cilobradine. Ligands of smaller size, for their part, are lodged in a hydrophobic cavity and form stable bonds with the proteins. Our results seem to suggest that blocking with Ivabradine is more effi- cient but that the stable bonds of small ligands have a greater potential for better affinity. However, the observed interactions suggest that the specificity towards isoforms would come from the kinetics of the channels and state dependencies of ligands rather than identified interactions. Finally, the binding of the ligands to the closed conformation of the HCN1 channel suggests that there would be a closed-bound conformation that is not known since no ligand has been able to access the pore.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study evaluated the anesthetic efficacy of eugenol and essential oils (EOs) of Aloysia triphylla and Lippia alba on aquaculture, as well as their stress-reducing effect during transport of shrimp and fish. The effectiveness of eugenol on blocking noxious stimulis was also tested. Sub-adult and post-larvae of white shrimp (Litopenaeus vannamei), albine and grey silver catfish (Rhamdia quelen) and fat snook (Centropomus paralellus) were evaluated for anesthetic induction time and recovery during short and long exposure to the EOs and eugenol. Oxidative stress parameters were also evaluated in sub-adult shrimps. Both silver catfish strains and fat snook were transported in water containing EO of A. triphylla at different concentrations and the reduction of stress was evaluated. An experimental model was elaborated to test the analgesic activity of eugenol in zebrafish (Danio rerio). The concentrations of eugenol, EO of A. triphylla and L. alba recommended for anesthesia in shrimps were respectively 200, 300 and 750 μL L-1 for sub-adults and 175, 300 and 500 μL L-1 for post-larvae. The concentrations indicated for transport are between 20-50, 20-30 and 50 μL-1 for sub adults and for post-larvae only eugenol and EO of A. triphylla at 20 and 20-50 μL L-1. A better antioxidant capacity in the hemolymph was obtained with 30 μL L-1 EOs of A. triphylla and L. alba and 20 μL L-1 eugenol. Albine silver catfish juveniles induced and recovered form anesthesia in longer time than grey ones and 200 μL L-1 was considered the most suitable concentration for both strains. In the transport the OE de A. triphylla increased whole body cortisol but did not promote ion loss. The increase of the concentration of OE A. triphylla proportionately decreased and increased the time of induction and recovery from anesthesia in fat snooks. The concentration of 20 μL L-1 reduced blood glucose levels after 24 h and plasma cortisol and lactate levels after 6h. Eugenol itself did not presented analgesic activity, but after the injection of acetic acid in the anterior portion eugenol blocked the noxious reflex. / O trabalho objetivou avaliar a eficácia anestésica do eugenol e óleos essenciais (OEs) de Aloysia triphylla e Lippia alba em aquicultura, bem como a eficácia dos mesmos como redutores de estresse durante o transporte de camarões e peixes e verificar efeito bloqueador de estímulos nocivos do eugenol. Camarões (sub-adultos e pós larvas), ambas variedades de jundiá e robalos-peva foram transportados em água contendo óleo essencial de A. triphylla em diferentes concentrações e avaliada a redução do estresse. Para eugenol foi traçado modelo experimental de avaliação da atividade analgésica em peixe-zebra (Danio rerio). As concentrações de eugenol e OE de A. triphylla e L. alba recomendadas para anestesia em camarões foram, respectivamente de 200, 300,750 μL L-1 para sub-adultos e para pós-larvas 175, 300 e 500 μL L-1. Para transporte concentrações entre 20-50; 20-30 e 50 μL L-1 são apropriadas para sub-adultos e para pós-larvas somente eugenol e OE de A. triphylla 20 e 20-50 μL L-1. Uma melhor capacidade antioxidante na hemolinfa de camarões foi obtida com 30 μL L-1 OEs de A. triphylla e L. alba e 20 μL L-1 para eugenol. Juvenis albinos de jundiá induziram e recuperaram da anestesia em maior tempo que os cinzas e considerou-se 200 μL L-1 a concentração mais adequada. No transporte, embora o OE de A. triphylla tenha causado aumento de cortisol corporal em jundiás, houve redução da perda de íons. Em robalospeva o aumento da concentração do OE de A. triphylla proporcionalmente diminuiu e aumentou o tempo de indução e recuperação anestésica, respectivamente. A concentração de 20 μL L-1 reduziu os níveis de glicose sanguínea após 24h e de cortisol e lactato após 6h. O eugenol por si só não apresentou atividade analgésica, porém quando injetado ácido acético na porção anterior de zebrafish a resposta ao efeito nocivo foi bloqueada.