Le système cholinergique est impliqué dans les phénomènes d’attention, de mémoire et d’apprentissage et les récepteurs cholinergiques régulent de multiples fonctions du système nerveux central. Néanmoins, leur rôle au niveau de la modulation des propriétés du cortex visuel reste à être établi. L’un des objectifs de cette thèse était d’étudier le rôle des récepteurs muscariniques impliqués dans le fonctionnement normal du cortex visuel. Nous avons pu déterminer que les récepteurs muscariniques sont impliqués dans l’établissement de nombreuses propriétés visuelles telles la taille des champs récepteurs, la sensibilité au contraste, la sélectivité à la fréquence spatiale et la finesse de la connectivité corticale. L’autre objectif était d’identifier les récepteurs cholinergiques impliqués dans la potentiation des capacités visuelles. Nous avons amélioré le traitement cognitif de l’information visuelle par stimulation électrique du télencéphale basal (noyau où sont localisés les corps cellulaires cholinergiques) et par la stimulation cholinergique par le donépézil, un inhibiteur de l’acétylcholinestérase. La combinaison répétée d’une stimulation visuelle et cholinergique (qu’elle soit électrique ou pharmacologique) améliore similairement l’activité corticale visuelle. Toutefois, les récepteurs impliqués ne sont pas les mêmes. Suite à la stimulation pharmacologique, ce sont principalement les récepteurs muscariniques qui influencent l’acuité visuelle de manière tardive et cette modulation est plus précoce lors de la stimulation électrique. Ces résultats démontrent que le couplage répétitif d’une stimulation cholinergique et d’une stimulation visuelle est en mesure d’améliorer l’activité corticale visuelle. Le fait de connaître les récepteurs cholinergiques impliqués permettra dans un futur proche de les cibler directement pour améliorer la fonction corticale. / The cholinergic system is involved in attention, learning and memory and cholinergic receptors regulate multiple functions of the central nervous system. Nevertheless, their role in modulating the properties of the visual cortex remains to be established. One of the objectives of this thesis was to study the role of muscarinic receptors involved in the normal function of the visual cortex. We have been able to determine that the muscarinic receptors are involved in the establishment of many visual properties such as the size of the receptor fields, contrast sensitivity, spatial frequency selectivity and accuracy of the cortical connectivity. The other objective was to identify the cholinergic receptors involved in the potentiation of visual abilities. We improved the cognitive processing of visual information by electrical stimulation of the basal forebrain (the nucleus where the cholinergic cell bodies are located) and by cholinergic stimulation using donepezil, an acetylcholinesterase inhibitor. The repeated combination of visual and cholinergic stimulations (whether electrical or pharmacological) similarly enhances visual cortical activity. However, the receptors involved are not the same. Following the pharmacological stimulation, it is mainly the muscarinic receptors that influence visual acuity with a delay in the receptors expression and this modulation is earlier for the electrical stimulation. These results demonstrate that repetitive coupling of cholinergic stimulation and visual stimulation can enhance visual cortical activity. Knowing the cholinergic receptors involved will allow in a near future to target them directly to improve cortical function.

Using a high throughput closed respirometry method to measure oxygen consumption, I determined metabolic rates in asexual and sexual Schmidtea mediterranea and Girardia dorotocephala, as a function of temperature, taxon, stressors, reproductive mode, age, regeneration, and specific dynamic action. This study has shown that oxygen consumption can reliably be measured in planaria using optode closed respirometry, and also provided a reliable method for measuring wet mass in planaria, which has been a challenge to researchers in the past. This research revealed that oxygen consumption in S. mediterranea is 1.5-2.1X greater in the sexual strain over the asexual strain at 13-18°C. Within the sexual strain, oxygen consumption is 1.5 -2.2X greater in sexually mature adults over the sexually immature groups (hatchlings, juveniles, and regenerating sexuals). Furthermore, I was able to quantify differences in sexual morphology between these groups exhibiting significant differences in oxygen consumption. The results of this research supports a theory of higher metabolic costs with sexual maturity in S. mediterranea. Therefore, this study has established sexual and asexual S. mediterranea as simple, yet attractive models for investigating energetic costs between sexual and asexual phenotypes. This research also provided quantitative values for specific dynamic action in planaria, with a maximum increase in oxygen consumption of 160% induced by feeding, as well as metabolic relationships in planaria involving temperature, age, and regeneration. These values establish planaria as one of the simplest animal models in which common metabolic patterns, such as SDA and poikilothermic temperature sensitivity, have been demonstrated. Therefore, this research has contributed to the overall knowledge of the basic physiology in this animal, providing the framework for future metabolic studies in planaria involving environmental factors, reproduction, regeneration, development, and aging. Information from this study may supplement interpretation and understanding of modern cellular, molecular, and genomic studies in planaria.

Le réseau neuronal de l’hippocampe joue un rôle central dans la mémoire en modifiant de façon durable l’efficacité de ses synapses. Dans les interneurones de la couche oriens/alveus (O/A), l’induction de la potentialisation à long terme (PLT) requiert les courants postsynaptiques excitateurs évoqués par les récepteurs métabotropes du glutamate de sous-type 1a (CPSEmGluR1a) et l’entrée subséquente de Ca2+ via des canaux de la famille des transient receptor potential (TRP). Le but de ce projet était d’identifier les canaux TRP responsables des CPSEmGluR1a et d’explorer les mécanismes moléculaires régulant leur ouverture. Nous avons déterminé par des enregistrements électrophysiologiques que les CPSEmGluR1a étaient spécifiques aux interneurones O/A et qu’ils étaient indépendants de la phospholipase C. Nous avons ensuite examiné l’expression des TRPC et leur interaction avec mGluR1a par les techniques de RT-PCR, d’immunofluorescence et de co-immunoprécipitation. Nos résultats montrent que TRPC1 et mGluR1a s’associent dans l’hippocampe et que ces deux protéines sont présentes dans les dendrites des interneurones O/A. En revanche, TRPC4 ne semble s’associer à mGluR1a qu’en système recombinant et leur colocalisation paraît limitée au corps cellulaire. Finalement, nous avons procédé à des enregistrements d’interneurones dans lesquels l’expression des TRPC a été sélectivement supprimée par la transfection d’ARN interférant et avons ainsi démontré que TRPC1, mais non TRPC4, est une sous-unité obligatoire du canal responsable des CPSEmGluR1a. Ces travaux ont permis de mieux comprendre les mécanismes moléculaires à la base de la transmission synaptique des interneurones O/A et de mettre en évidence un rôle potentiel de TRPC1 dans la PLT. / The hippocampal neuronal network plays a crucial role in memory by producing long lasting changes in the efficacy of its synapses. In interneurons of stratum oriens/alveus (O/A), long term potentiation (LTP) induction requires metabotropic glutamate receptor subtype 1a (mGluR1a)-evoked excitatory postsynaptic currents (EPSCs) and subsequent Ca2+ entry through transient receptor potential (TRP) channels. The objectives of this project were to identify the TRP channels that mediate mGluR1a-evoked EPSCs and to explore molecular mechanisms that underlie their activation. Electrophysiological recordings showed that mGluR1a-evoked EPSCs were specifically observed in O/A interneurons and they were phospholipase C-independent. We then examined TRPC expression and their interaction with mGluR1a by RT-PCR, immunofluorescence and co-immunoprecipitation techniques. Our results show that TRPC1 and mGluR1a associate in hippocampus and that both proteins have overlapping distributions in dendrites of O/A interneurons. In contrast, TRPC4 seems to associate with mGluR1a only in recombinant system and their co-localization appears to be limited to the cell body. Finally, we performed recordings of interneurons in which TRPC expression was selectively suppressed by small interfering RNAs and we found that TRPC1, but not TRPC4, is an obligatory subunit of the channel that mediate mGluR1a-evoked EPSCs. This work brought new insight on molecular mechanisms underlying synaptic transmission of O/A interneurons and uncovered a potential role for TRPC1 in LTP.

Le réseau neuronal de l’hippocampe joue un rôle central dans la mémoire en modifiant de façon durable l’efficacité de ses synapses. Dans les interneurones de la couche oriens/alveus (O/A), l’induction de la potentialisation à long terme (PLT) requiert les courants postsynaptiques excitateurs évoqués par les récepteurs métabotropes du glutamate de sous-type 1a (CPSEmGluR1a) et l’entrée subséquente de Ca2+ via des canaux de la famille des transient receptor potential (TRP). Le but de ce projet était d’identifier les canaux TRP responsables des CPSEmGluR1a et d’explorer les mécanismes moléculaires régulant leur ouverture. Nous avons déterminé par des enregistrements électrophysiologiques que les CPSEmGluR1a étaient spécifiques aux interneurones O/A et qu’ils étaient indépendants de la phospholipase C. Nous avons ensuite examiné l’expression des TRPC et leur interaction avec mGluR1a par les techniques de RT-PCR, d’immunofluorescence et de co-immunoprécipitation. Nos résultats montrent que TRPC1 et mGluR1a s’associent dans l’hippocampe et que ces deux protéines sont présentes dans les dendrites des interneurones O/A. En revanche, TRPC4 ne semble s’associer à mGluR1a qu’en système recombinant et leur colocalisation paraît limitée au corps cellulaire. Finalement, nous avons procédé à des enregistrements d’interneurones dans lesquels l’expression des TRPC a été sélectivement supprimée par la transfection d’ARN interférant et avons ainsi démontré que TRPC1, mais non TRPC4, est une sous-unité obligatoire du canal responsable des CPSEmGluR1a. Ces travaux ont permis de mieux comprendre les mécanismes moléculaires à la base de la transmission synaptique des interneurones O/A et de mettre en évidence un rôle potentiel de TRPC1 dans la PLT. / The hippocampal neuronal network plays a crucial role in memory by producing long lasting changes in the efficacy of its synapses. In interneurons of stratum oriens/alveus (O/A), long term potentiation (LTP) induction requires metabotropic glutamate receptor subtype 1a (mGluR1a)-evoked excitatory postsynaptic currents (EPSCs) and subsequent Ca2+ entry through transient receptor potential (TRP) channels. The objectives of this project were to identify the TRP channels that mediate mGluR1a-evoked EPSCs and to explore molecular mechanisms that underlie their activation. Electrophysiological recordings showed that mGluR1a-evoked EPSCs were specifically observed in O/A interneurons and they were phospholipase C-independent. We then examined TRPC expression and their interaction with mGluR1a by RT-PCR, immunofluorescence and co-immunoprecipitation techniques. Our results show that TRPC1 and mGluR1a associate in hippocampus and that both proteins have overlapping distributions in dendrites of O/A interneurons. In contrast, TRPC4 seems to associate with mGluR1a only in recombinant system and their co-localization appears to be limited to the cell body. Finally, we performed recordings of interneurons in which TRPC expression was selectively suppressed by small interfering RNAs and we found that TRPC1, but not TRPC4, is an obligatory subunit of the channel that mediate mGluR1a-evoked EPSCs. This work brought new insight on molecular mechanisms underlying synaptic transmission of O/A interneurons and uncovered a potential role for TRPC1 in LTP.

Events around conception such as maternal undernutrition and twinning may have effects on offspring physiology and disease risk in adulthood. Periconceptional undernutrition alters offspring physiology and adult pathology without affecting birth size, while twinning affects birth size and physiology but with inconsistent effects on adult pathology. We investigated the effects of these two periconceptional events and their interaction on maternal cardiovascular adaptation to pregnancy and fetal growth, physiology and endocrinology in late gestation in sheep. Pre and/or postconception undernutrition resulted in increased uterine blood flow in late gestation, but no change in maternal blood volume. Preconception undernutrition alone resulted in a relatively large placenta with a small, slow-growing fetus in late gestation. In contrast, postconception undernutrition alone resulted in a fetus with rapid late-gestation growth that was maintained through a maternal fast. Fetuses of ewes undernourished throughout both periods were similar in growth rate and size to controls. Maternal fasting also demonstrated that plasma levels of C-type natriuretic peptide are acutely and independently regulated by nutrient supply in mother and fetus. Fetuses of ewes undernourished both pre- and postconception had increased glucose disposal following a glucose challenge. Hypothalamic-pituitary-adrenal axis tests in these fetuses showed decreased pituitary adrenocorticotropin hormone response to direct stimulation but increased adrenal response to decreased cortisol negative feedback. Twin fetuses grew more slowly in late gestation than singletons. Twins also had a smaller insulin response to arginine and a greater insulin response to glucose, but periconceptional undernutrition abolished this difference. Twins had suppressed baseline hypothalamic-pituitary-adrenal axis function and decreased adrenal sensitivity compared to singletons, but increased fetal pituitary adrenocorticotropin hormone response to direct stimulation and decreased cortisol negative feedback. These studies suggest that firstly, fetal size is a poor reflection of fetal growth trajectory, physiology and endocrinology. Secondly, pre- and postconception undernutrition affect late-gestation fetal growth in different ways, while undernutrition in both periods alters fetal endocrine status in late gestation. Thirdly, the biology of twin fetal development is fundamentally different from that of singletons, which may explain the inconsistency of the relationship between birth weight and adult disease risk in twins. / Auckland Medical Research Foundation, Health Research Council of New Zealand

Events around conception such as maternal undernutrition and twinning may have effects on offspring physiology and disease risk in adulthood. Periconceptional undernutrition alters offspring physiology and adult pathology without affecting birth size, while twinning affects birth size and physiology but with inconsistent effects on adult pathology. We investigated the effects of these two periconceptional events and their interaction on maternal cardiovascular adaptation to pregnancy and fetal growth, physiology and endocrinology in late gestation in sheep. Pre and/or postconception undernutrition resulted in increased uterine blood flow in late gestation, but no change in maternal blood volume. Preconception undernutrition alone resulted in a relatively large placenta with a small, slow-growing fetus in late gestation. In contrast, postconception undernutrition alone resulted in a fetus with rapid late-gestation growth that was maintained through a maternal fast. Fetuses of ewes undernourished throughout both periods were similar in growth rate and size to controls. Maternal fasting also demonstrated that plasma levels of C-type natriuretic peptide are acutely and independently regulated by nutrient supply in mother and fetus. Fetuses of ewes undernourished both pre- and postconception had increased glucose disposal following a glucose challenge. Hypothalamic-pituitary-adrenal axis tests in these fetuses showed decreased pituitary adrenocorticotropin hormone response to direct stimulation but increased adrenal response to decreased cortisol negative feedback. Twin fetuses grew more slowly in late gestation than singletons. Twins also had a smaller insulin response to arginine and a greater insulin response to glucose, but periconceptional undernutrition abolished this difference. Twins had suppressed baseline hypothalamic-pituitary-adrenal axis function and decreased adrenal sensitivity compared to singletons, but increased fetal pituitary adrenocorticotropin hormone response to direct stimulation and decreased cortisol negative feedback. These studies suggest that firstly, fetal size is a poor reflection of fetal growth trajectory, physiology and endocrinology. Secondly, pre- and postconception undernutrition affect late-gestation fetal growth in different ways, while undernutrition in both periods alters fetal endocrine status in late gestation. Thirdly, the biology of twin fetal development is fundamentally different from that of singletons, which may explain the inconsistency of the relationship between birth weight and adult disease risk in twins. / Auckland Medical Research Foundation, Health Research Council of New Zealand

Events around conception such as maternal undernutrition and twinning may have effects on offspring physiology and disease risk in adulthood. Periconceptional undernutrition alters offspring physiology and adult pathology without affecting birth size, while twinning affects birth size and physiology but with inconsistent effects on adult pathology. We investigated the effects of these two periconceptional events and their interaction on maternal cardiovascular adaptation to pregnancy and fetal growth, physiology and endocrinology in late gestation in sheep. Pre and/or postconception undernutrition resulted in increased uterine blood flow in late gestation, but no change in maternal blood volume. Preconception undernutrition alone resulted in a relatively large placenta with a small, slow-growing fetus in late gestation. In contrast, postconception undernutrition alone resulted in a fetus with rapid late-gestation growth that was maintained through a maternal fast. Fetuses of ewes undernourished throughout both periods were similar in growth rate and size to controls. Maternal fasting also demonstrated that plasma levels of C-type natriuretic peptide are acutely and independently regulated by nutrient supply in mother and fetus. Fetuses of ewes undernourished both pre- and postconception had increased glucose disposal following a glucose challenge. Hypothalamic-pituitary-adrenal axis tests in these fetuses showed decreased pituitary adrenocorticotropin hormone response to direct stimulation but increased adrenal response to decreased cortisol negative feedback. Twin fetuses grew more slowly in late gestation than singletons. Twins also had a smaller insulin response to arginine and a greater insulin response to glucose, but periconceptional undernutrition abolished this difference. Twins had suppressed baseline hypothalamic-pituitary-adrenal axis function and decreased adrenal sensitivity compared to singletons, but increased fetal pituitary adrenocorticotropin hormone response to direct stimulation and decreased cortisol negative feedback. These studies suggest that firstly, fetal size is a poor reflection of fetal growth trajectory, physiology and endocrinology. Secondly, pre- and postconception undernutrition affect late-gestation fetal growth in different ways, while undernutrition in both periods alters fetal endocrine status in late gestation. Thirdly, the biology of twin fetal development is fundamentally different from that of singletons, which may explain the inconsistency of the relationship between birth weight and adult disease risk in twins. / Auckland Medical Research Foundation, Health Research Council of New Zealand

Events around conception such as maternal undernutrition and twinning may have effects on offspring physiology and disease risk in adulthood. Periconceptional undernutrition alters offspring physiology and adult pathology without affecting birth size, while twinning affects birth size and physiology but with inconsistent effects on adult pathology. We investigated the effects of these two periconceptional events and their interaction on maternal cardiovascular adaptation to pregnancy and fetal growth, physiology and endocrinology in late gestation in sheep. Pre and/or postconception undernutrition resulted in increased uterine blood flow in late gestation, but no change in maternal blood volume. Preconception undernutrition alone resulted in a relatively large placenta with a small, slow-growing fetus in late gestation. In contrast, postconception undernutrition alone resulted in a fetus with rapid late-gestation growth that was maintained through a maternal fast. Fetuses of ewes undernourished throughout both periods were similar in growth rate and size to controls. Maternal fasting also demonstrated that plasma levels of C-type natriuretic peptide are acutely and independently regulated by nutrient supply in mother and fetus. Fetuses of ewes undernourished both pre- and postconception had increased glucose disposal following a glucose challenge. Hypothalamic-pituitary-adrenal axis tests in these fetuses showed decreased pituitary adrenocorticotropin hormone response to direct stimulation but increased adrenal response to decreased cortisol negative feedback. Twin fetuses grew more slowly in late gestation than singletons. Twins also had a smaller insulin response to arginine and a greater insulin response to glucose, but periconceptional undernutrition abolished this difference. Twins had suppressed baseline hypothalamic-pituitary-adrenal axis function and decreased adrenal sensitivity compared to singletons, but increased fetal pituitary adrenocorticotropin hormone response to direct stimulation and decreased cortisol negative feedback. These studies suggest that firstly, fetal size is a poor reflection of fetal growth trajectory, physiology and endocrinology. Secondly, pre- and postconception undernutrition affect late-gestation fetal growth in different ways, while undernutrition in both periods alters fetal endocrine status in late gestation. Thirdly, the biology of twin fetal development is fundamentally different from that of singletons, which may explain the inconsistency of the relationship between birth weight and adult disease risk in twins. / Auckland Medical Research Foundation, Health Research Council of New Zealand

<p> Down syndrome (DS) is a complex genetic disorder caused by the triplication of human chromosome 21 (Hsa21). The presence of an extra copy of an entire chromosome greatly disrupts the copy number and expression of over 350 protein coding genes. This gene dosage imbalance has far-reaching effects on normal development and aging, leading to cognitive and skeletal defects that emerge earlier in life than the general population.</p> <p> The present study begins by characterizing skeletal development in young male Ts65Dn mice to test the hypothesis that skeletal defects in male Ts65Dn mice are developmental in nature.Femurs from young mice ranging from postnatal day 12- to 42-days of age (P12-42) were measured and analyzed by microcomputed tomography (μCT). Cortical defects were present generally throughout development, but trabecular defects emerged at P30 and persisted until P42. </p> <p> The gene <i>Dual-specificity tyrosine-regulated kinase 1a </i>(<i>Dyrk1a</i>) is triplicated in both DS and in Ts65Dn mice and has been implicated as a putative cause of both cognitive and skeletal defects. To test the hypothesis that trisomic <i>Dyrk1a</i> is related to the emergence of trabecular defects at P30, expression of <i>Dyrk1a</i> in the femurs of male Ts65Dn mice was quantified by qPCR. Expression was shown to fluctuate throughout development and overexpression generally aligned with the emergence of trabecular defects at P30.</p> <p> The growth rate in trabecular measures between male Ts65Dn and euploid littermates was similar between P30 and P42, suggesting a closer look into cellular mechanisms at P42. Assessment of proliferation of BMSCs, differentiation and activity of osteoblasts showed no significant differences between Ts65Dn and euploid cellular activity, suggesting that the cellular microenvironment has a greater influence on cellular activity than genetic background.</p> These data led to the hypothesis that reduction of <i>Dyrk1a</i> gene expression and pharmacological inhibition of DYRK1A could be executed during a critical period to prevent the emergence of trabecular defects at P30. To tests this hypothesis, doxycycline-induced cre-lox recombination to reduce <i>Dyrk1a</i> gene copy number or the DYRK1A inhibitor CX-4945 began at P21. The results of both genetic and pharmacological interventions suggest that trisomic <i>Dyrk1a</i> does not influence the emergence of trabecular defects up to P30. Instead, data suggest that the critical window for the rescue of trabecular defects lies between P30 and P42.

L'athérosclérose est la principale cause de maladies coronariennes, affectant les artères de grand et moyen calibre. C'est une maladie inflammatoire chronique caractérisée par des plaques situées dans la couche de l’intima, composées de cellules inflammatoires, de cellules musculaires lisses, de composants fibreux et de lipides. Qu'il provienne de source alimentaire ou hépatique, le cholestérol qui s'accumule dans les macrophages des tissus périphériques, comme la paroi artérielle, engendre une réaction inflammatoire et doit être conséquemment mobilisé à l'aide d’accepteurs de cholestérol comme les lipoprotéines de haute densité (HDL). Ce processus spécifique est appelé transport inverse du cholestérol (mRCT). Des études ont démontré que l'apolipoprotéine A-I (apoA-I) pourrait être un acteur clé dans la régulation du mRCT, exerçant des effets différents de ceux du HDL. Plus important encore, le système lymphatique a récemment été identifié comme un nouvel acteur essentiel dans l'élimination du cholestérol de la lésion athérosclérotique (Martel et al., JCI 2013). Il a été démontré que sans vaisseaux lymphatiques fonctionnels, la mobilisation du cholestérol hors de la plaque ne peut pas être réalisée correctement et aggrave la maladie. Le réseau lymphatique est parallèle au système sanguin et il est présent dans presque tous les tissus du corps. C'est un acteur essentiel dans le maintien de l'homéostase des fluides, dans le transport des cellules immunitaires de la périphérie vers les ganglions lymphatiques correspondants, ainsi que dans l’absorption des lipides alimentaires de l'intestin vers la circulation sanguine. Le système lymphatique comprend les vaisseaux lymphatiques (LVs) initiaux et collecteurs, ainsi que les ganglions lymphatiques, qui ont une anatomie spécifique et des rôles distincts. La lymphe, le liquide clair qui circule dans les LVs, se jette dans la circulation sanguine au niveau de la veine sous-clavière. Les plaquettes sont responsables de la régulation de cette séparation des vaisseaux sanguins et lymphatiques via la formation d’un thrombus formé lors de l’interaction de leur récepteur CLEC-2 avec la podoplanine présente sur les cellules endothéliales lymphatiques. Il a également été démontré que l’activité plaquettaire était nécessaire tout au long de la vie pour maintenir l’intégrité des jonctions des LVs. L'athérosclérose est également caractérisée par une activation cellulaire et une apoptose accrue. Par conséquent, ces activités cellulaires peuvent entraîner la formation de particules submicroniques appelées vésicules extracellulaires qui ont des effets variables, mais souvent néfastes, sur l'endothélium sanguin et l'évolution de la plaque. La maladie cardiovasculaire a été associée à une augmentation du nombre des vésicules extracellulaires (EVs) en circulation, et nous croyons que ces véhicules pourraient être impliqués dans le dysfonctionnement lymphatique lié à l'athérosclérose. D'après des données récentes publiées au cours de ma maîtrise, l'amélioration du transport lymphatique pourrait limiter la progression de l'athérosclérose et favoriser la régression de la plaque. Nous avons montré que le transport lymphatique est altéré chez les jeunes souris prédisposés à développer l'athérosclérose, même avant l'apparition de la plaque. Nous avons prouvé que cet effet est d’abord associé à un défaut au niveau des vaisseaux collecteurs et nous suggérons que l'amélioration de la liaison du VEGF-C/ VEGFR3 puisse supprimer ce défaut spécifique. L'objectif global de cette thèse était de poursuivre dans cette voie et de mieux définir le rôle de l’important facteur de croissance lymphatique, VEGF-C, et de la lipoprotéine apoA-I dans la maintenance de l’intégrité et la fonction des vaisseaux lymphatiques. En outre, une meilleure description des composants de la lymphe, en particulier des agents libérés par les cellules, a été jugée nécessaire. La première publication nous a permis de montrer que, lorsqu'elles étaient injectées avec un mutant du facteur de croissance VEGF-C ciblant spécifiquement le récepteur VEGFR-3 (VEGF-C 152s), avant l'administration d'une diète pro-athérogène, les souris Ldlr-/- étaient protégées contre l’accumulation excessive dans la plaque et celle-ci était plus stable à long terme. La capacité de contraction soutenue des vaisseaux lymphatiques collecteurs et l'expression accrue de VEGFR-3 et de FOXC2 observée chez ces souris traitées avec VEGF-C-152s ont contribué à la clairance des composants nocifs contenus dans les tissus périphériques tels que les macrophages et le cholestérol. La deuxième publication a montré que des souris Ldlr-/- athérosclérotiques traitées à faible dose avec de l’apoA-I, présentaient un transport lymphatique accru et une hyperperméabilité des vaisseaux lymphatiques collecteurs abrogée, possiblement par une modulation de l’activité plaquettaire. La troisième publication est la première à démontrer la présence de vésicules extracellulaires d'origines hétérogènes dans la lymphe des souris et que le nombre de différents sous-types augmente chez les souris athérosclérotiques. Collectivement, ces études confirment la présence d'un dysfonctionnement lymphatique chez la souris avant même l'apparition de la plaque, et il est intéressant de noter que ce dysfonctionnement est principalement associé à un défaut des vaisseaux lymphatiques collecteurs, limitant ainsi le transport de la lymphe des tissus périphériques vers le sang. Différents traitements avec des facteurs de croissance et des lipoprotéines peuvent potentiellement moduler l’apparition et la progression de la lésion en améliorant la fonction lymphatique à différents stades de la maladie athérosclérotique. Nos découvertes concernant la présence de EVs dans la lymphe représentent leur potentiel en tant que biomarqueurs, mais également une nouvelle cible pour mieux comprendre la dysfonction lymphatique. / Atherosclerosis is the principal cause of coronary artery disease (CAD), affecting large- and medium-sized arteries. It is a chronic inflammatory disease characterized by intimal plaques composed of inflammatory cells, smooth muscle cells, fibrous components and lipids. Cholesterol that accumulates within macrophages in peripheral tissues, like the arterial wall, whether from dietary or synthetic sources, promotes inflammatory responses and needs to be excreted with the help of the cholesterol acceptor high density lipoprotein (HDL). This specific process is termed macrophage reverse cholesterol transport (mRCT) and studies have demonstrated that lipid free apolipoprotein A-I (apoA-I) could be a key player in mRCT regulation, exuding different effects than HDL. More importantly, recently, the lymphatic system has been identified as a novel prerequisite player in the removal of cholesterol out of the atherosclerotic lesion (Martel et al., JCI 2013). It has been demonstrated that without functioning lymphatic vessels cholesterol mobilization from the plaque cannot be properly achieved and aggravates the disease. The lymphatic network runs in parallel to the blood vasculature and is present in almost all the tissues of the body. It is a crucial player in maintaining fluid homeostasis, trafficking immune cells from the periphery to corresponding lymph nodes, as well as transporting lipids from the intestine to the circulation. The lymphatic system comprises the initial and collecting lymphatic vessels (LVs), as well as lymph nodes, all with a specific anatomy and distinctive roles. Lymph, the clear fluid that circulates within LVs drains towards the bloodstream at the level of the subclavian vein. Platelets are responsible to regulate this blood/lymphatic vessel separation by forming a clog, upon the interaction of their C-type lectin-like receptor 2 (CLEC-2) with podoplanin, present on lymphatic endothelial cells. Platelet activity has also been shown to be required throughout life in order to maintain LV junction integrity. Atherosclerosis is also characterized by increased cellular activation and apoptosis. Consequently, these cellular activities may result in the formation of submicron particles called extracellular vesicles (EVs) that have variable effects on the blood endothelium and subsequent plaque evolution. CAD has been associated with increased circulating EVs, and we suspect that these EVs might be involved in atherosclerosis-related lymphatic dysfunction. Based on recent data collected during my master’s degree, there is evidence that enhancing lymphatic transport could limit atherosclerosis progression and favour plaque regression. We showed that lymphatic transport is impaired in young, atherosclerosis-prone mice, even before atherosclerosis onset. We believe it to be potentially associated with a defect in the lymphatic pumping capacity, and we suggest that enhancing VEGF-C/VEGFR-3 binding can abolish this specific defect. The global objective of this thesis was to pursue along this path and better delineate the role of the important lymphatic-specific growth factor, VEGF-C and the lipoprotein apoA-I, on collecting LVs function. Furthermore, a better understanding of lymph components, especially cellular releasants was deemed necessary. The first publication allowed us to show that when injected with VEGF-C 152s, before the administration of a pro-atherogenic regimen, Ldlr-/- mice were protected from excessive plaque formation and long-term, had a more stable plaque. The sustained contraction capacity of the collecting lymphatic vessels and the enhanced expression of VEGFR-3 and FOXC2 observed in these VEGF-C-152s treated mice contributed to the clearance of harmful components contained in peripheral tissues such as the macrophages and cholesterol. The second publication showed that atherosclerotic Ldlr-/- mice treated with low-dose lipid-free apoA-I had enhanced lymphatic transport and abrogated collecting LV permeability possibly through modulation of platelet activity. The third publication is the first ever to demonstrate the presence of extracellular vesicles of heterogeneous origins in the lymph of mice, and that their levels differ in atherosclerosis. Collectively, these studies confirm that lymphatic dysfunction is present before the onset of atherosclerosis, and particularly of interest, that this dysfunction is primarily associated with a defect in the collecting vessels, thereby limiting the lymph transport from peripheral tissues to the blood. Different treatments with growth factors and lipoproteins have the potential to modulate the lesion onset and progression through the enhancement of lymphatic function, while our findings regarding the presence of EVs in lymph represents their potential as biomarkers, but also a new venue to better understand lymphatic dysfunction.