La fusariose des épis, causée majoritairement par le champignon pathogène Fusarium graminearum (Fg), est une des principales maladies du blé tendre (Triticum aestivum). Pendant son cycle infectieux, l’agent pathogène produit des mycotoxines appartenant principalement aux trichothécènes de type B, tel que le déoxynivalénol (DON), qui sont toxiques pour l’homme et l’animal. Plusieurs loci à caractère quantitatif (QTLs) impliqués dans la résistance à la fusariose des épis ont été identifiés. Certains ont été corrélés avec une détoxication de la mycotoxine, principalement par conjugaison du DON en DON-3-O-glucose (D3G), une réaction enzymatique catalysée par des UDP-glucosyltransferases (UGT). Néanmoins, peu d’études ont conduit des analyses fonctionnelles dans des plantes hôtes de la maladie afin de relier directement la glucosylation de la mycotoxine avec la résistance à la maladie in planta, et aucune d’entre elles n'a été effectuée sur des gènes de détoxication du blé tendre. Notre équipe, à l'aide de la céréale modèle Brachypodium distachyon, a démontré que l'UGT Bradi5g03300 est capable de conférer une tolérance au DON par glucosylation en DON 3-O-glucose et qu’elle est impliquée dans l'établissement précoce d'une résistance quantitative à la fusariose des épis. Le présent travail avait pour objectif de transférer les analyses fonctionnelles menées sur la céréale modèle Brachypodium distachyon au blé tendre. Dans une première approche, le gène Bradi5g03300 a été introduit dans la variété de blé Apogée, sensible à la fusariose. Les analyses phénotypiques effectuées sur les lignées de blé transgéniques exprimant constitutivement le gène Bradi5g03300, montrent une résistance plus élevée à la maladie ainsi qu'une tolérance à la mycotoxine par rapport à la lignée contrôle. Parallèlement, en utilisant une approche de synténie entre les génomes de B. distachyon et du blé tendre, nous avons identifié un gène de blé, orthologue au gène Bradi5g03300.La transformation de l’écotype de B. distachyon sensible à la fusariosepar ce gène candidat a été effectuée pour déterminer rapidement sa capacité à conjuguer le DON in planta et son implication dans la résistance à la fusariose. En conclusion, ce projet contribue à accroître les connaissances concernant la relation fonctionnelle entre la glucosylation DON et la résistance à la fusariose dans le blé tendre et à fournir des gènes candidats à inclure dans les processus de sélection. / Fusarium head blight (FHB) caused by fungi of the Fusarium genus is a widespread disease of wheat (Triticum aestivum) and other small-grain cereal crops. The main causal agent of FHB, Fusarium graminearum, can produce mycotoxins mainly belonging to type B trichothecenes, such as deoxynivalenol (DON) that can negatively affect humans, animals and plants. Several quantitative trait loci (QTLs) for resistance to FHB have been identified some of which have been correlated with efficient DON detoxification, mainly through the conjugation of DON into DON-3-O-glucose (D3G), a reaction catalyzedby UDP-glucosyltransferases (UGTs). Nevertheless, only few studies have conducted functional analyses to directly correlate DON glucosylation and resistance in planta and none were performed on wheat UGT gene(s). Our team, using the model cereal species Brachypodium distachyon, has recently demonstrated that the Bradi5g03300 UGT is able to confer tolerance to DON following glucosylation of DON into DON 3-O-glucose and is involved in the early establishment of quantitative resistance to FHB. In the present work, we transferred the functional analyses conducted on the model species Brachypodium distachyon to bread wheat. In a first approach the B. distachyon Bradi5g03300 gene has been introduced through biolistic-mediated transformation in the wheat variety Apogee, susceptible to FHB. The phenotypic analyses conducted on homozygous transgenic wheat constitutively expressing the Bradi5g03300 gene showed that they exhibit higher resistance to FHB as well as increased root tolerance to DON compared to the control line. In parallel, using a synteny approach between B. distachyon and bread wheat genomes we identified a wheat candidate gene orthologous to the B. distachyon Bradi5g03300 gene. This wheat gene after validation through gene expression pattern during wheat infection, was introduced by transformation into B. distachyon to rapidly determine its ability to conjugate DON into D3G in planta and its involvement in FHB resistance. In conclusion, this project contributes to increase the knowledge concerning the functional relationship between DON glucosylation and FHB resistance in wheat and provide candidate genes to include in selection processes.

La morphine constitue l'un des analgésiques les plus utilisés en milieu hospitalier pour soulager les douleurs aiguës et chroniques. Elle exerce ses effets analgésiques en se liant aux récepteurs opioïdes μ (MOR) présents au niveau central et périphérique et possède des effets secondaires, incluant la tolérance, qui limitent son usage à long terme. La première partie de mes travaux avait pour objectif d'étudier le phénomène de tolérance à la morphine et d'en déterminer les mécanismes sous-jacents. A l'heure actuelle, ce phénomène est décrit comme résultant d'une désensibilisation des MOR conduisant à leur endocytose. Nos résultats montrent qu'en cas de tolérance, le catabolisme de la morphine est exacerbé au niveau du système nerveux central. Mes travaux ont également porté sur la caractérisation de la Créatine Kinase (CK) comme étant une protéine liant la morphine à très haute affinité. Nos résultats montrent que la CK possède 2 sites de liaison pour la morphine avec des affinités similaires. En étudiant l'effet potentiel de la CK sur l'analgésie induite par la morphine in vivo, nous avons mis en évidence, que les peptides correspondant aux 2 sites de liaison à la morphine étaient par eux-mêmes analgésiques et que cette analgésie semblait dépendre des récepteurs opioïdes. / Morphine is one of the most used analgesics in hospitals to relieve acute and chronic pain. Morphine exerts its analgesic effects by binding central and peripheral μ opioid receptors (MORs) and has many side effects that limit its long-term use including tolerance. The first part of my thesis was aimed to study the phenomenon of morphine tolerance and to determine the underlying mechanisms. Previously, this phenomenon was explained as resulting !rom MORs desensitization by endocytosis. However, our results show that in case of tolerance, the catabolism of morphine is exacerbated in the central nervous system in particular in astocytes. The second part of my work has focused on the characterization of the Creatine Kinase (CK) as a novel protein that binds morphine with a high affinity. Our results showed that CK has two binding sites with a similar affinity for morphine. Surprisingly, by studying the potential effect of CK on morphine-induced analgesia in vivo, we noticed that the two peptides corresponding to morphine binding sites are analgesics and such analgesia seems to be mediated by opioid receptors.

<p>This investigation focused on the mRNA expression of drug metabolising Cytochromes P-450 (CYP) and UDP-glucuronosyltransferases (UGT) and the transport protein P-glycoprotein (Pgp) in the small intestine of humans and rats.</p><p>The mRNA expression of the investigated genes in the human small intestine (duodenum) varies between individuals giving each one of us personal profile. In general, the most dominant forms are Pgp, CYPs 2C9, 2D6, 3A4, and UGTs 1A1, 1A10, 2B7. However, which of these is the highest expressed one varies between individuals.</p><p>The correlation in expression between some CYP forms and UGT forms respectively is relatively high, which indicates that they have some regulatory mechanisms in common. It was also shown that the mRNA expression of both CYPs and UGTs may be affected by endogenous and exogenous factors. Sex and ethnic background, affected the mRNA expression of CYP2A6 and 2E1 respectively. Commonly used drugs such as acetylsalicylicacid (ASA) and omeprazole (omep) affect CYP2A6, CYP2E1 (ASA) and CYP3A4, UGT1A4 (omep). The expression of UGT1A4 is also affected by smoking. All these factors are commonly used and can therefore lead to important drug-drug interactions.</p><p>It was also shown that the human small intestinal CYP mRNA expression pattern differs from that found in the rat. The rat CYP expression is rather constant between the different individuals, and the main rat intestinal forms are CYP1A1, CYP2C, CYP2D6 and CYP3A1. The expression is the same for females and males and no difference can be seen between the different segments of the rat small intestine. As metabolic studies have often been done with rat liver we compared the mRNA expression in the two organs. We found that the mRNA expression of 1A1 was absent in the liver and that the CYP2B1, CYP2Cs, CYP2D1 and Pgp all had a stronger mRNA expression in the small intestine compared to the liver. It is therefore important to realise that results from metabolic studies on liver may not be directly extrapolated to the small intestine.</p><p>Artemisinin is an orally used drug in multidrug treatment of malaria in Southeast Asia. It has been suggested that artemisinin can induce drug metabolism and therefore be involved in drug-drug interactions. This study shows that artemisinin induces mainly the CYP2B via nuclear receptor CAR.</p>

This investigation focused on the mRNA expression of drug metabolising Cytochromes P-450 (CYP) and UDP-glucuronosyltransferases (UGT) and the transport protein P-glycoprotein (Pgp) in the small intestine of humans and rats. The mRNA expression of the investigated genes in the human small intestine (duodenum) varies between individuals giving each one of us personal profile. In general, the most dominant forms are Pgp, CYPs 2C9, 2D6, 3A4, and UGTs 1A1, 1A10, 2B7. However, which of these is the highest expressed one varies between individuals. The correlation in expression between some CYP forms and UGT forms respectively is relatively high, which indicates that they have some regulatory mechanisms in common. It was also shown that the mRNA expression of both CYPs and UGTs may be affected by endogenous and exogenous factors. Sex and ethnic background, affected the mRNA expression of CYP2A6 and 2E1 respectively. Commonly used drugs such as acetylsalicylicacid (ASA) and omeprazole (omep) affect CYP2A6, CYP2E1 (ASA) and CYP3A4, UGT1A4 (omep). The expression of UGT1A4 is also affected by smoking. All these factors are commonly used and can therefore lead to important drug-drug interactions. It was also shown that the human small intestinal CYP mRNA expression pattern differs from that found in the rat. The rat CYP expression is rather constant between the different individuals, and the main rat intestinal forms are CYP1A1, CYP2C, CYP2D6 and CYP3A1. The expression is the same for females and males and no difference can be seen between the different segments of the rat small intestine. As metabolic studies have often been done with rat liver we compared the mRNA expression in the two organs. We found that the mRNA expression of 1A1 was absent in the liver and that the CYP2B1, CYP2Cs, CYP2D1 and Pgp all had a stronger mRNA expression in the small intestine compared to the liver. It is therefore important to realise that results from metabolic studies on liver may not be directly extrapolated to the small intestine. Artemisinin is an orally used drug in multidrug treatment of malaria in Southeast Asia. It has been suggested that artemisinin can induce drug metabolism and therefore be involved in drug-drug interactions. This study shows that artemisinin induces mainly the CYP2B via nuclear receptor CAR.