No description available.

Tyrosine nitration results in altered function of smooth muscle voltage-gated L-type calcium channel. We explored the possibility that smooth muscle contains denitrase activity to allow functional recovery of the calcium channel without requiring synthesis of new channel proteins. Following peroxynitrite treatment of mouse colonic smooth muscle strips, CaCl2 (1 mM)-induced smooth muscle contraction was significantly reduced by 67% (P ≤ 0.05), which reversed by approximately 86% upon periodic washing within 2 hr period (P ≤ 0.001). The effect of the c-Src kinase inhibitor, PP2, on muscle contraction was also restored after 2 hr post-peroxynitrite treatment consistent with the thesis that recovery from tyrosine nitration allows for tyrosine phosphorylation of the calcium channel. In addition, sodium orthovanadate prevented nitration-induced inhibition of muscle contraction by approximately 90%. Moreover, denitration of nitrated proteins was observed by western blots in smooth muscle cells over 2 hr. Since nitrotyrosine formation interferes with tyrosine kinase pathways involved in cell signaling, the presence of denitrase activity in smooth muscle cells may have profound and important effects in restoring the function of nitrated proteins involved in cell signaling processes.

In the heart, the primary role of the L-type calcium channel (LTCC) CaV1.2 is to conduct calcium into cardiomyocytes and initiate contraction. However, part of the CaV1.2 channel itself, the cleaved carboxyl-terminus (CCt) can also localize to the nucleus and regulate gene transcription. Therefore, the goal of this dissertation project was to determine the role and regulation of CCt in the embryonic and adult heart. The global hypothesis of my dissertation project is that CCt localizes to the nucleus in embryonic and adult cardiomyocytes via a calcium-mediated mechanism and regulates transcription. A model of pharmacological LTCC block-induced perturbation of murine embryonic heart development was first utilized to study the role of CCt. Pharmacological block at embryonic day 10 perturbed cardiogenesis and increased CaV1.2 expression. This result was not mimicked by removal of extracellular calcium or inhibition of calcium release from the sarcoplasmic reticulum. Co-currently, pharmacological block decreased CCt nuclear localization in embryonic cardiomyocytes. At the transcriptional level, CCt suppressed the CaV1.2 promoter. This indicated that the observed upregulation of CaV1.2 induced by pharmacological block may be caused by nuclear localization of the transcriptional repressor, CCt. Therefore, the conclusion was made that pharmacological LTCC block perturbed embryonic cardiogenesis by decreasing nuclear localization of the transcription factor CCt; implying a role for CCt in embryonic heart development. Next, CCt regulation was studied in the adult heart. Similar to the embryonic heart, pharmacological LTCC block decreased nuclear localization of CCt. Inhibition of the calcium activated phosphatase calcineurin also decreased CCt nuclear localization. To determine a role for CCt in the adult heart, CCt nuclear localization was measured in response to hypertrophic stimuli. Serum-induced cardiomyocyte hypertrophy significantly increased nuclear localization of CCt. In conclusion, this dissertation supports the hypothesis that CCt localizes to the nucleus in embryonic and adult cardiomyocytes, and that this regulation is mediated by calcium entry into the cardiomyocyte. Furthermore, data from this dissertation suggests that CCt nuclear localization may play an important role in embryonic heart development and adult cardiac hypertrophy.

Sudden cardiac death (SCD) is a leading cause of fatalities. Several methods have been developed to predict instability in myocytes which could lead to SCD. The focus of this study was on altering memory in myocytes, i.e. hysteresis in restitution of action potential duration (APD), by differing levels of calcium. Determination of alteration was implemented by using a diastolic interval (DI) control program that implements a sinusoidal change in DI. Plotting APD versus previous DI, i.e. restitution, produces a hysteresis loop. From these hysteresis loops, five parameters were used to determine measures of memory: area, thickness, overall tilt, max delay and min delay. Calcium levels were then altered with either verapamil or BAPTA-AM. Statistically significant effects were found for the verapamil study, but not for the BAPTA-AM study. Simulations were used to explain significant results. The verapamil findings support clinical studies that have shown verapamil to not have anti-arrhythmic effects. Theory predicts that a decrease in memory would decrease the stability of a system, and perhaps verapamil may not increase stability as hypothesized previously. The results of the BAPTA-AM study were inconclusive, and further investigation is needed before it can be determined that BAPTA-AM has no significant effect on memory.

The L-type calcium channel (LTCC) provides trigger Ca2+ for sarcoplasmic reticulum Ca2+-release and LTCC function is influenced by interacting proteins including the LTCC Distal Carboxyl-terminus (DCT) and calmodulin. DCT is proteolytically cleaved, and re-associates with the LTCC complex to regulate calcium channel function. DCT reduces LTCC barium current (IBa,L) in reconstituted channel complexes, yet the contribution of DCT to ICa,L in cardiomyocyte systems is unexplored. This study tests the hypothesis that DCT attenuates cardiomyocyte ICa,L. We measured LTCC current and Ca2+ transients with DCT co-expressed in murine cardiomyocytes. We also heterologously co-expressed DCT and CaV1.2 constructs with truncations corresponding to the predicted proteolytic cleavage site, CaV1.2Δ1801, and a shorter deletion corresponding to well-studied construct, CaV1.2Δ1733. DCT inhibited IBa,L in cardiomyocytes, and in HEK 293 cells expressing CaV1.2Δ1801 and CaV1.2Δ1733. Ca2+-CaM relieved DCT block in cardiomyocytes and HEK cells. The selective block of IBa,L combined with Ca2+-CaM effects suggested that DCT-mediated blockade may be relieved under conditions of elevated Ca2+. We therefore tested the hypothesis that DCT block is dynamic, increasing under relatively low Ca2+, and show that DCT reduced diastolic Ca2+ at low stimulation frequencies but spared high frequency Ca2+-entry. DCT reduction of diastolic Ca2+ and relief of block at high pacing frequencies, and under conditions of supraphysiological bath Ca2+ suggests that a physiological function of DCT is to increase the dynamic range of Ca2+ transients in response to elevated pacing frequencies. Our data motivates the new hypothesis that DCT is a native reverse use-dependent inhibitor of LTCC current.

Abstract The dihydropyridine receptor (DHPR) in the skeletal muscle plasma membrane functions as a voltage sensor for excitation-contraction coupling. In the present work the expression and special features of DHPR were studied under various conditions. In order to localize and visualize the DHPRs, a method using fluorophore-conjugated dihydropyridine molecules as a probe was developed. In addition, different laboratory assays and electrophysiological measurements were used to study the expression of the myofibrillar proteins, force production of the muscle and conduction velocity of the plasma membrane. During the postnatal development of mouse skeletal muscle the density of DHPR increased. By the time of DHPR appearance, the expression of sodium channels had started and the typical assembly of basic structural components and membrane compartments was clearly detectable. According to the histochemical analysis, the DHPR was selectively expressed in type IIA muscle fibres of mouse. In addition to the fibre type specificity, the uneven distribution of DHPRs was also seen at the muscle level. The attenuation of the contraction force after addition of DHPR blocker was largest in muscles with a high percentage of type IIA fibres. In fish muscles, the distribution of DHPR was homogenous and the density between different fibre types was quite similar. In gastrocnemius muscle of mouse, the density and mRNA expression of DHPR increased significantly by 21.5 and 66.8%, respectively, after a 15-week aerobic exercise programme. The increase correlated significantly with the raise in % myosin heavy chain IIa isoform. In thigh and heart muscles, no significant changes were observed. In fish, the environmental change following hatchery release and downstream migration of 46.5 km induced an increase in the DHPR density in swimming muscles. Furthermore, a transition of phenotypic profile from fast-to-slow fibres was observed. Taken together, the present data provide evidence for the fact that the expression of DHPR increases during postnatal development. Moreover, the expression correlates with a specific fibre-type metabolism, thus having an impact on the overall contractile properties of the muscle. This is further manifested as an increase in the DHPR density after endurance training in mammalian as well as in fish muscle. In addition, a strong correlation exists between the level of muscle activity and the density of DHPR. / Tiivistelmä Poikkijuovaisen lihaksen solukalvolla esiintyvä dihydropyridiini (DHP) reseptori toimii jännitesensorina lihaksen ärsytys-supistus kytkennässä. Tässä työssä tutkittiin DHP reseptorin erityispiirteitä sekä erilaisten fysiologisten tekijöiden vaikutusta reseptorin ilmenemiseen. DHP reseptorien visualisointiin kehitettiin histologinen värjäysmenetelmä, jossa merkkiaineena toimi fluorofori-konjugoitu DHP molekyyli. Lisäksi työssä tutkittiin lihaksen proteiinien ekspressiota, lihaksen voimantuottoa sekä solukalvon johtonopeutta erilaisten määritysmenetelmien ja elektrofysiologisten mittausmenetelmien avulla. Tulokset osoittivat, että yksilönkehityksen aikana DHP reseptorien tiheys kasvoi hiiren poikkijuovaisessa lihaksessa. DHP reseptorien ekspression alkaessa natriumkanavat olivat jo ilmaantuneet solukalvolle ja lihassolun rakenne oli pitkälle erilaistunut. Histokemiallisten määritysten perusteella DHP reseptorin ekspressio oli selektiivistä. Reseptoreita esiintyi runsaimmin tyypin IIA soluissa. Reseptoriproteiinin solutyyppispesifisyys oli huomattavissa myös lihastasolla. Tulosten mukaan proteiinin salpaaja alensi lihaksen supistumisvoimaa erityisesti niissä hiiren lihaksissa, joiden solutyyppikoostumuksessa tyypillä IIA on suuri prosentuaalinen osuus. Kalan lihaksissa DHP reseptorit olivat homogeenisesti jakautuneet. Lisäksi reseptoritiheys oli samankaltainen eri solutyyppien välillä. 15 viikon aerobinen harjoittelu lisäsi sekä DHP reseptorin proteiini- että mRNA- ekspressiota tilastollisesti merkitsevästi (21,5 ja 66,8 %) hiiren kantalihaksessa. Ekspression kasvu korreloi merkitsevästi samanaikaisesti tapahtuneen myosiini isomuoto IIa määrän kasvun kanssa. Reisi- ja sydänlihaksessa merkittäviä muutoksia ei havaittu. Kalan uintilihaksissa DHP reseptorien tiheys kasvoi vapauttamisen jälkeisen ympäristön muutoksen ja 46,5 km pituisen vaelluksen jälkeen. Lisäksi lihasten solutyyppikoostumuksessa tapahtui muutos kohti hitaasti supistuvia solutyyppejä. Yhteenvetona voidaan todeta, että saatujen tulosten perusteella DHP reseptorien ekspressio kasvaa syntymän jälkeen hiiren poikkijuovaisessa lihaksessa. Solujen erilaistuessa ekspressio korreloi solutyyppimetabolian kanssa vaikuttaen edelleen lihaksen supistumisominaisuuksiin. Tästä johtuen myös kestävyysharjoittelun seurauksena DHP reseptorien määrä kasvaa sekä nisäkkään että kalan lihaksissa. Erityisesti lihaksen aktiivisuudella on merkitystä DHP reseptorin ekspressioon.

Modelos matemáticos de motoneurônios têm sido desenvolvidos para auxiliar na compreensão dos fenômenos que envolvem o sistema neuromuscular. Entretanto, a maioria dos modelos já desenvolvidos baseou-se na premissa de que a árvore dendrítica tem um comportamento passivo, o que ocorre em animais anestesiados, mas pode não ocorrer durante o comportamento motor normal de um animal intacto. Experimentos com animais descerebrados, em que as vias monoaminérgicas encontravam-se ativas, mostraram que os motoneurônios podem apresentar comportamentos mais complexos decorrentes da presença de condutâncias iônicas voltagem-dependentes que se situam nos dendritos e são responsáveis pela gênese de uma corrente de entrada persistente. Nesse sentido, um primeiro objetivo deste trabalho foi o de desenvolver novos modelos matemáticos de motoneurônios de diferentes tipos (i.e. dos tipos S, FR e FF), computacionalmente eficientes e contendo em seus compartimentos dendríticos uma condutância de cálcio do tipo L, de forma que os fenômenos de biestabilidade, potencial platô e amplificação da corrente sináptica efetiva possam ser gerados. Um segundo objetivo foi o de verificar como a presença da condutância iônica ativa no dendrito influencia o comportamento motoneuronal quando o mesmo está sujeito a entradas sinápticas de diferentes tipos. Os novos modelos foram parametrizados baseando-se em dados da literatura experimental para motoneurônios de gatos descerebrados e validados segundo os protocolos experimentais básicos que permitem caracterizar cada tipo de modelo como sendo totalmente ou parcialmente biestável. As entradas sinápticas foram simuladas por processos pontuais de Poisson e os trens de potenciais de ação dos motoneurônios foram analisados. Uma modulação senoidal da intensidade do processo pontual foi usada para estimar as respostas em frequência de cada modelo. Observou-se que, funcionalmente, a presença da condutância iônica dendrítica pode favorecer a ação do motoneurônio durante tarefas posturais, pois, uma vez ativada, a corrente de entrada persistente eleva a excitabilidade motoneuronal tornando os disparos mais regulares, além de prover uma alta sensibilidade dos modelos a entradas sinápticas de baixa frequência, correspondentes às oscilações observadas durante a manutenção da postura ereta quieta. / Mathematical models of motoneurons have been developed as an aid to the understanding of phenomena involving the neuromuscular system, but most of these models have been based on the hypothesis of a passive dendritic tree. This holds for anesthetized animals but not necessarily during normal motor behavior of the intact animal. Experiments with decerebrate animals in which the monoaminergic tracts were maintained intact have shown that more complex behaviors may emerge in motoneurons due to dendritic voltage-gated ionic conductances, which are responsible for a persistent inward current. Therefore, the first aim of this work was to develop computationally-efficient new motoneuron models of different types (i.e. type S, FR and FF) that include a dendritic L-type calcium conductance so that bistability, plateau potential and enhancement of effective synaptic current may be generated. The second aim of this research was to evaluate the effects of the active dendritic ionic conductance on the input-output mapping of presynaptic to postsynaptic spike trains. The new models were parameterized based on data reported in experimental literature on the decerebrate cat preparation, and they were validated using appropriate protocols for either fully or partially bistable dynamics. The synaptic inputs were simulated by Poisson point processes and the output spike trains were analyzed. Sinusoidal modulation of the point process intensity was used for the estimation of each models frequency response. The results suggested that an active dendritic ionic conductance in motoneurons has a functional role during postural tasks, because, when activated, the persistent inward current enhances the motoneuronal excitability, reducing the variability of interspike intervals, and focusing the sensitivity of the models to low frequency inputs that correspond to the low-frequency oscillations that typically occur during quiet standing posture.

Ces dernières décennies ont mis à jour une implication pathologique nouvelle del’aldostérone, via le récepteur aux minéralocorticoïdes (RM) dans le coeur. L’ensemble desdonnées issues des études expérimentales et des essais cliniques suggère une association délétèreentre l’aldostérone et la survenue d’arythmies. L’utilisation d’antagonistes du RM prévient cesarythmies. Cependant, les voies de signalisations, comme les mécanismes moléculaires soustendantces effets bénéfiques du blocage des RM demeurent incertains. Nous avons accumulésdes preuves d’une modulation de la signalisation calcique dans le cardiomyocyte, et en particulierde l’influx calcique (Ca2+) au travers du canal Ca2+ de type L (LTCC). Celui-Ci pourrait être unecible primaire de l’aldostérone et du RM dans les cardiomyocytes ventriculaires. Toutefois, lesmécanismes par lesquels l’aldostérone et le RM régulent l’expression du LTCC restent à définir.Au cours de ces travaux menés sur cardiomyocytes de rats nouveau-Nés, nous avonsétudiés les évènements moléculaires par lesquels l’aldostérone exerce ses effets sur le CaV1.2,qui correspond à la sous-Unité principale du LTCC formant le pore du canal ; cette protéine estcodée par le CACNA1C. Par microscopie confocale, nous avons suivi en temps réel le traffickingnucléo-Cytoplasmique du RM couplé à la GFP en réponse à l’aldostérone, démontrant ainsi queles RM cardiaques sont fonctionnels. Le traitement durant 24 heures des cardiomyocytes avec del’aldostérone montre une augmentation dose-Dépendante des protéines et de l’ARN messager duCaV1.2. L’utilisation de la technique du gène rapporteur de la luciférase permet l’analyse del’activité du promoteur du CaCNA1C. Celui-Ci montre une activité transcriptionnelle dose ettemps dépendante en réponse à l’aldostérone. De plus, ces effets sont dépendant des RM carinhibés en présence de RU28318, un antagoniste sélectif du RM, ou par l’utilisation de siRNAdirigés contre le RM. L’analyse in silico de la séquence du promoteur du CaCNA1C nous a permisd’identifier cinq séquences putatives correspondant à des éléments de réponse auxglucocorticoïdes (GRE). La mutation du site le plus lointain du site d’initiation de la transcriptionne révèle aucun changement dans les réponses transcriptionnelles induites par un RM humainconstitutivement actif (hMRΔ5,6) ou dans les réponses doses-Dépendantes de l’aldostérone ou dela déxaméthasone, un glucocorticoïde de synthèse. La mutation des trois sites GRE putatifssuivants provoque une diminution des réponses au hMRΔ5,6 comme à l’aldostérone, alors que lesréponses à la déxaméthasone sont soit inchangées, soit augmentées. En contraste, la mutation dusite le plus proximal du promoteur augmente de façon importante l’activité transcriptionnelle dupromoteur en réponse au hMRΔ5,6, à l’aldostérone comme à la déxaméthasone.Ces résultats démontrent que le LTCC cardiaque constitue une cible directe del’aldostérone et du RM, et apportent de nouvelles perspectives quant aux conséquencesmoléculaires et fonctionnelles engendrées par l’activation délétère du système minéralocorticoïdedans la défaillance cardiaque. / During the past decades, major novel pathogenic roles of the steroid hormone,aldosterone, via the Mineralocorticoid Receptor (MR) have been identified in heart. Collectively,experimental studies and clinical trials, suggest a detrimental association between aldosteroneand life threatening arrhythmias that may be prevented by MR blockade. However, the signalingpathways and underlying mechanisms still remain elusive. We have accumulated evidence thatmodulation of Ca2+ signaling, especially Ca2+ influx via L-Type Ca2+ channel (LTCC), might bethe primary aldosterone/MR target in ventricular cardiomyocytes. Yet, the molecularmechanisms by which MR regulates expression of LTCC remain to be defined. Here, weinvestigated, in primary cultures of neonatal rat ventricular myocytes, the molecular eventscritical for aldosterone-Mediated cardiac effects on CaV1.2, the pore-Forming main subunit ofLTCC, which is encoded by the CaCNA1C gene.We showed that cardiac MR are functional as demonstrated by aldosterone-Induced MRnucleocytoplasmic trafficking observed by time-Lapse imaging of transfected GFP-Labeled MRusing confocal microscopy. Aldosterone exposure for 24 hours, induced a dose-Dependentincrease in CaV1.2 expression at both mRNA and protein levels. Analysis of the CaCNA1Cpromoter activity using luciferase reporter assays, revealed a dose- and time-Dependent activationby aldosterone. These effects were inhibited in the presence of either RU28318, a selective MRantagonist, or MR siRNA. In silico analyze enabled us to identify five putative GlucocorticoidResponse Elements (GRE) within the CaCNA1C promoter sequence. The mutation of the mostdistal GRE from Transcription Start Site (TSS) did not altered responses either elicited by theconstitutively active human MR (hMRΔ5,6) or dose-Dependent effects of aldosterone anddexamethasone (a synthetic glucocorticoïd with minimal MR effect). Mutations of the three nextones decreased responses to hMRΔ5,6 and aldosterone, whereas dexamethasone responses wereeither unchanged or increased. In sharp contrast, the mutation of the most proximal GRE fromTSS, increased responses to hMRΔ5,6, aldosterone and dexamethasone.These results provide new insights into the molecular mechanisms associated with cardiacMR activation, and suggest that LTCC is a primary MR target, with subsequent molecular andfunctional consequences that could lead to MR-Related cardiac dysfunction.

Modelos matemáticos de motoneurônios têm sido desenvolvidos para auxiliar na compreensão dos fenômenos que envolvem o sistema neuromuscular. Entretanto, a maioria dos modelos já desenvolvidos baseou-se na premissa de que a árvore dendrítica tem um comportamento passivo, o que ocorre em animais anestesiados, mas pode não ocorrer durante o comportamento motor normal de um animal intacto. Experimentos com animais descerebrados, em que as vias monoaminérgicas encontravam-se ativas, mostraram que os motoneurônios podem apresentar comportamentos mais complexos decorrentes da presença de condutâncias iônicas voltagem-dependentes que se situam nos dendritos e são responsáveis pela gênese de uma corrente de entrada persistente. Nesse sentido, um primeiro objetivo deste trabalho foi o de desenvolver novos modelos matemáticos de motoneurônios de diferentes tipos (i.e. dos tipos S, FR e FF), computacionalmente eficientes e contendo em seus compartimentos dendríticos uma condutância de cálcio do tipo L, de forma que os fenômenos de biestabilidade, potencial platô e amplificação da corrente sináptica efetiva possam ser gerados. Um segundo objetivo foi o de verificar como a presença da condutância iônica ativa no dendrito influencia o comportamento motoneuronal quando o mesmo está sujeito a entradas sinápticas de diferentes tipos. Os novos modelos foram parametrizados baseando-se em dados da literatura experimental para motoneurônios de gatos descerebrados e validados segundo os protocolos experimentais básicos que permitem caracterizar cada tipo de modelo como sendo totalmente ou parcialmente biestável. As entradas sinápticas foram simuladas por processos pontuais de Poisson e os trens de potenciais de ação dos motoneurônios foram analisados. Uma modulação senoidal da intensidade do processo pontual foi usada para estimar as respostas em frequência de cada modelo. Observou-se que, funcionalmente, a presença da condutância iônica dendrítica pode favorecer a ação do motoneurônio durante tarefas posturais, pois, uma vez ativada, a corrente de entrada persistente eleva a excitabilidade motoneuronal tornando os disparos mais regulares, além de prover uma alta sensibilidade dos modelos a entradas sinápticas de baixa frequência, correspondentes às oscilações observadas durante a manutenção da postura ereta quieta. / Mathematical models of motoneurons have been developed as an aid to the understanding of phenomena involving the neuromuscular system, but most of these models have been based on the hypothesis of a passive dendritic tree. This holds for anesthetized animals but not necessarily during normal motor behavior of the intact animal. Experiments with decerebrate animals in which the monoaminergic tracts were maintained intact have shown that more complex behaviors may emerge in motoneurons due to dendritic voltage-gated ionic conductances, which are responsible for a persistent inward current. Therefore, the first aim of this work was to develop computationally-efficient new motoneuron models of different types (i.e. type S, FR and FF) that include a dendritic L-type calcium conductance so that bistability, plateau potential and enhancement of effective synaptic current may be generated. The second aim of this research was to evaluate the effects of the active dendritic ionic conductance on the input-output mapping of presynaptic to postsynaptic spike trains. The new models were parameterized based on data reported in experimental literature on the decerebrate cat preparation, and they were validated using appropriate protocols for either fully or partially bistable dynamics. The synaptic inputs were simulated by Poisson point processes and the output spike trains were analyzed. Sinusoidal modulation of the point process intensity was used for the estimation of each models frequency response. The results suggested that an active dendritic ionic conductance in motoneurons has a functional role during postural tasks, because, when activated, the persistent inward current enhances the motoneuronal excitability, reducing the variability of interspike intervals, and focusing the sensitivity of the models to low frequency inputs that correspond to the low-frequency oscillations that typically occur during quiet standing posture.

Agelanthus dodoneifolius est une plante de la pharmacopée africaine utilisée en médecine traditionnelle pour le traitement de pathologies cardiovasculaires. Un fractionnement bioguidé a permis d'isoler une nouvelle dihydropyranone : baptisée Dodonéine (Ddn), elle possède des propriétés hypotensives et vasorelaxantes chez le rat.L'objectif de la thèse est de caractériser la ou les cible(s) moléculaire(s) de la Ddn dans le système cardiovasculaire et d'identifier un dérivé plus sélectif.Nos résultats montrent que la Ddn bloque le courant calcique de type L des cardiomyocytes ventriculaires de rat et des cellules musculaires lisses vasculaires, d'environ 30% avec une IC50 de l'ordre du µM. La Ddn apparaît comme un nouveau bloqueur des canaux calciques de type L avec des propriétés électrophysiologiques qui lui sont propres. Une étude biochimique a montré que l'anhydrase carbonique (CA) est également inhibée par la Ddn, et nous avons caractérisé l'expression de plusieurs isoformes au sein de muscle lisse vasculaire : CA II, III, XIII et XIV. Nos travaux précisent que leur inhibition augmente le pH intracellulaire, pouvant conduire à l'activation des canaux KCa. Ainsi, la Ddn induit un effet vasorelaxant en inhibant deux protéines: les canaux calciques et l'anhydrase carbonique. Nous avons ensuite démontré que les dérivés de la Ddn nouvellement synthétisés sont aussi vasorelaxants ; toutefois, le plus efficace, la Ddn-Bicyclique-OH, ne concurrence pas les bloqueurs calciques utilisés en clinique. Cependant, l'effet vasorelaxant de la Ddn et Ddn-Bc-OH caractérisé sur des artères systémiques humaines est en accord avec l'utilisation de la plante en médecine traditionnelle. / Agelanthus dodoneifolius is one of the medicinal plants used in African pharmacopeia and traditional medicine for the treatment of cardiovascular diseases. Bioguided fractionation has allowed isolating one of its main active principles named Dodoneine (Ddn). It is a new dihydropyranone which exerts hypotensive and vasorelaxant effects on rat.The aim of this work is to characterize the molecular(s) target(s) of Ddn on cardiovascular system and identify the most selective and effective Ddn's derivative.Electrophysiological studies revealed that Ddn blocks L-type calcium current density of about 30% with an IC50 value of about 2 µM on cardiac myocytes and on vascular smooth muscle cells. Ddn appears as a new natural calcium channel blocker which has its own electrophysiological properties. As it has been shown, by biochemical study, that carbonic anhydrase is a potential target for Ddn, we have demonstrated that isozymes II, III, XIII are present on vascular smooth muscle cells and inhibited by Ddn. This inhibition resulted in a rise in pHi of about 0.31, leading to KCa channel activation. Interestingly, Ddn induced vasorelaxation by targeting two proteins: calcium channel and carbonic anhydrase. Then, we demonstrated that Ddn's derivatives newly synthetized have also vasorelaxant properties via the inhibition of L-type calcium current. Among them, Ddn-Bicyclique-OH is the most effective but appears not to be a better pharmacological tool compared to the calcium channel blockers already use in clinical. However, in accordance to the use of the plant in traditional medicine, our results clearly show that Ddn and Ddn-Bc-OH induce a vasorelaxant effect on human systemic artery.