Les avancées récentes dans la technique de transgénèse utilisant l’approche AAV ont permis de générer de nouveaux modèles animaux. Depuis quelques années, le développement des modèles de la maladie de Parkinson (MP) a amélioré la compréhension des mécanismes physiopathologiques de ce trouble dégénératif. Cependant, aucun modèle mammifère ne reproduit à ce jour la neurodégénérescence liée à l’âge associée à la pathologie synucléine et la symptomatologie motrice et non motrice. L’objectif de mon travail de thèse fut de développer de nouvelles stratégies de transgénèse chez le rat en utilisant ces vecteurs viraux pour la modélisation de la MP. Le challenge est de parvenir à une infection virale la plus précoce possible afin de transduire un maximum de neurones dopaminergiques. Pour cela, différentes voies d’approche ont été testées pour améliorer le processus de transgénèse : 1) l’injection de vecteurs viraux dans le rete testis de jeunes mâles pour transduire les cellules de la lignée germinale, 2) l’injection dans les embryons pré-implantatoires, 3) l’injection in utero intracérébroventriculaire et enfin 4) l’injection intracardiaque au premier jour de vie des animaux. Parmi elles, les injections in utero et intracardiaques ont permis d’aboutir à une transduction importante d’un grand nombre de structures cérébrales avec un tropisme neuronal. Ces techniques ouvrent la voie à la génération de nouveaux modèles animaux de maladies à composante génétique et à la thérapie génique. / Recent advances in the transgenesis technique using the AAV approach have led to the generation of new animal models. In recent years, the development of models of Parkinson's disease (PD) has improved understanding of the pathophysiological mechanisms of this degenerative pathology. However, no mammalian model recapitulates the required age-dependant parkinsonian degeneration, the α-synuclein inclusion pathology and motor and non-motor symptoms. The objective of my Ph.D work was to develop new transgenesis strategies in rats using these viral vectors for modeling PD. The challenge is to achieve a viral infection as early as possible in order to transduce as many dopaminergic neurons as possible. To this end, different strategies have been tested to improve transgenesis efficacy : i) injection in rete testis in young male to transduce germinal cells, ii) injection into early stage embryos, iii) in utero intracerebroventricular injection and iv) intracardiac injection in one day-old animals. Among them, in utero and intracardiac injections led to neuronal transgene expression in most regions of the brain. These techniques pave the way for the generation of new animal models of genetic diseases and offer unique opportunities for gene therapy.

Retard de croissance intra-utérin (RCIU) et prééclampsie (PE) sont des complications majeures de la grossesse humaine et sont le plus souvent due à une insuffisance de vascularisation utéro-placentaire. Notre objectif était d'évaluer l’angiographie Doppler énergie tridimensionnelle (PDA) comme nouvel outil de dépistage de la PE et du RCIU et d’étude de la fonction placentaire et du RCIU sur modèles animaux. La corrélation entre les indices Doppler 3D et l'écoulement de sang réel au sein de l'unité utéro-placentaires a d'abord été évaluée sur modèle de brebis gravide, ainsi que l'impact des réglages de la machine. Un degré de corrélation plus élevé a été observé pour VI et VFI (r = 0,86 et 0,82 respectivement, p <0,0001) que pour FI (r = 0,64, p <0,0001).L'intérêt de la technique a été ensuite démontré dans un modèle de RCIU chez le lapin (femelles traitées par du L-NAME).Troisièmement, la valeur prédictive du PDA comme test de dépistage du RCIU et de la PE a été démontrée par la réalisation d'une étude prospective multicentrique chez 70 femmes enceintes à bas risque (AUC 0,95, 100% VAN avec une spécificité de 85% pour un seuil de 36.784 FI placentaire). / Intra-uterine growth retardation (IUGR) and preeclampsia (PE) are major complications of human pregnancy & are most often due to an insufficient utero-placental vascularization. Our aim was to evaluate the three-dimensional power Doppler angiography (PDA) as a new tool for the screening IUGR & PE & for the study placental function and IUGR in animal models. The correlation between 3D Doppler indices and the real blood flow within the utero-placental unit was first evaluated in the pregnant sheep model, as well as the impact of machine settings. A higher correlation degree was observed for VI and VFI (r = 0.86 and 0.82 respectively p<0.0001) than for FI (r = 0.64; p<0.0001).The interest of the technique was secondly demonstrated in a rabbit IUGR model (females treated with L-NAME).Thirdly, the predictive value of PDA as a screening test for IUGR & PE was demonstrated by conducting a prospective multicentric study in 70 low risk pregnant women (AUC 0.95, 100% NPV with a specificity of 85% for a 36.784 placental FI threshold).

Background: Exposure of skin to UVB radiation (290-320 nm) modulates the immune system, with most studies showing a suppression of Th1-driven immune responses. Investigations into the effects of UVB exposure on allergic respiratory responses have been limited. This study investigated the systemic effects of UVB on Th2-associated immune responses using two different murine models of allergic respiratory inflammation. The mechanism of immune regulation was also examined. Methods and Results: Two murine models of asthma were used: the papain model and the ovalbumin (OVA) model using papain and OVA, respectively, as the allergens. In the papain model, C57BL/6, histamine receptor-1 knockout (H1RKO) and histamine receptor-2 knockout (H2RKO) mice were exposed to a single 4 kJ/m2 dose of UVB (twice a minimal oedemal dose) on shaved dorsal skin three days prior to intranasal sensitisation with papain, a cysteine protease homologue of the house dust mite (Dermatophagoides pteronyssinus) allergen Der p 1. Sensitisation and boost each consisted of five daily intranasal doses of 1 µg papain whilst the challenge consisted of three daily intranasal doses of 100 µg papain. Asthmatic symptoms were assessed 24 h after the final challenge dose. H1RKO mice demonstrated enhanced papain-specific inflammatory responses in the lung-draining lymph nodes (LDLNs) whilst the responses of H2RKO mice closely mimicked those of C57BL/6 mice. UVB irradiation three days before sensitisation reduced in vitro papain-specific proliferation of LDLN cells from C57BL/6 and H1RKO mice but not H2RKO mice 24 h after challenge. The regulatory effect of UVB was transferred by adoptive transfer of 5 x 106 unfractionated LDLN cells from UVB-irradiated, papain-sensitised and -challenged C57BL/6 and H1RKO donor mice into naïve recipients of the corresponding strain that were ii subsequently sensitised and challenged with papain. Additionally, UVB exposure suppressed papain-induced IL-5 and IL-10 production in vitro by LDLN cells from H1RKO mice but not from C57BL/6 mice or H2RKO mice. The results of this study demonstrate systemic immunomodulation of responses to intranasally delivered antigen by UVB irradiation and the induction of regulatory cells in the LDLN following UVB exposure. Furthermore, these results implicate a role for the H2R in UVB-induced suppression of antigen-specific responses in the draining lymph nodes.

Parkinson&rsquo / s disease (PD) is characterized by the progressive loss of dopaminergic nigral neurons and striatal dopamine resulting in serious motor deficits but also some non-motor anomalies. Animal models of human neurodegenerative diseases are essential for better understanding their pathogenesis and developing efficient therapeutic tools. There are many different PD models, however, none of them is fully reproducing all the symptoms of the disease. In addition, different investigators use different behavioral measures which makes even more difficult to compare and evaluate the results. The aim of the present study was to compare motor and cognitive deficits in two most common models of PD: the Rotenone and 6-OHDA model, using a large battery of neurological tests and a probabilistic learning task. To the best of our knowledge, this is the first study to examine the effects of bilaterally induced Rotenone and 6-OHDA through behavioral test batteries assessing the cardinal motor symptoms and the cognitive abnormality of Parkinson&rsquo / s Disease in the same rat population. Also, the present study is unique on the basis of providing both longitudinal observations of behaviour in the same treatment group and the cross-sectional comparisons of the behavioural responses between different groups. In the current study, the neurotoxins were applied at relatively low doses of 3-4 &mu / g, bilaterally to the substantia nigra pars compacta (SNpc). Experiments were conducted on 50 young-adult male Sprague&ndash / Dawley rats randomly assigned to five experimental groups: Rotenone, 6-OHDA, vehicle (DMSO/Saline), and the intact control. The neurological tests included locomotor activity,catalepsy, rearing, stepping, and rotarod/accelerod tests. They were applied prior to, and on days 4-7-10-20-40-150 while the learning task was applied 49 days after drug infusion.During the first 2 postoperational months, both neurotoxins produced progressive deterioration in motor performance but showing no effect on cognitive functions. Five months after the surgery, regression of bradykinesia but persistence of sensorimotor deficits was noted. The tests&rsquo / results suggest different susceptibility of different motor functions to the degeneration of nigro-striatal (N-S) pathway. So, different tests were demonstrated to have different power in detecting similar motor deficits.

published_or_final_version / Anaesthesiology / Doctoral / Doctor of Philosophy

Stroke remains a leading cause of disability in the western world, with symptoms ranging in severity from mild congnitive or motor impairments, to severe impairments in both cognitive and motor domains. Despite ongoing research aimed at helping stroke patients the disease cannot be prevented or cured, therefore a large body of research has been aimed at identifying effective rehabilitative strategies. Based on our understanding of normal brain function, and the meachanisms mediating the limited spontaneous recovery that is observed following injury, factors that promote brain plasticity are likely to be effective treatments for stroke symptoms. The current thesis investigated three novel treatments (COX-2 inhibitor drug, vitamin supplement diet, and social experience) in a rat model of focal ischemia in the motor cortex. All three treatments have been previously shown to alter plasticity in the normal brain, however the current experiments show that the treatments have differential effects following stroke. The COX-2 inhibitors provided limited improvement in functional performance, whereas the vitamin supplement treatment had no effect. Social experience on the other hand was found to block the usually observed spontaneous improvements following the stroke. These results suggest that factors that alter dendritic plasticity may in fact serve as effective stroke treatments depending on the site and the mechanisms whereby the plastic changes are induced. / ix, 149 leaves : ill. ; 29 cm.

Middle cerebral artery (MCA) stroke can produce chronic incapacitating motor impairments. Understanding the neural basis of the motor syndromes is complicated by the diversity of neural structures damaged but the problem can be addressed in laboratory rats by inducing selective infarcts. Nevertheless, the motor syndromes that ensue from stroke in rats remain poorly understood and undermine its potential as a model for clinical stroke. The objective of the present thesis was to document the skilled reaching impairments from neocortical and subcortical MCA infarcts in rats. In addition, the integrity of the motor system components spared by the infarct was assessed neurophysiologically and neuroanatomically. Characteristic reaching impairments emerged from each infarct but there were also some overlapping features that might be explained by neural dysfunction extending beyond the boundaries of the infarct. The present studies showed that the laboratory rat is an ideal animal model for studying stroke, which should be of interest to both clinical and research scientists studying stroke. / xiii, 345 leaves : ill. ; 29 cm. + 1 CD-ROM

Ionizing radiation (IR), along with being an important diagnostic and treatment modality, is a potent tumor-causing agent, and the risk of secondary radiation treatment-related cancers is a growing clinical problem. Now some studies propose to link secondary radiation-induced cancers to an enigmatic phenomenon of bystander effects, whereby the exposed cells send signal damage and distress to their naïve neighbors and result in genome destabilization and carcinogenesis. Yet, no data existed on the bystander effects in an organ other than an exposed one. With this in mind, we focused on the analysis of existence and mechanisms of radiation-induced bystander effects in vivo. We have found that bystander effects occur in vivo in distant skin and spleen following half-body or cranial irradiation. These bystander effects resulted in elevated DNA damage, profound dysregulation of epigenetic machinery, and pronounced alterations in apoptosis, proliferation and gene expression. Bystander effects also exhibited persistency and sex specificity. The results obtained while using the animal model systems can potentially be extrapolated to different animals and humans. / xiii, 208 leaves : ill. ; 29 cm.

[Truncated abstract] Formation and consolidation of the retinotectal projection during optic nerve regeneration has been associated with two major interlinked processes. Initially, retinal ganglion cell (RGC) axons are guided by molecular guidance cues, such as the Eph receptor tyrosine kinases and their ligands, the ephrins, to their approximately correct location and form a coarse topographic map in the optic tecum. Such axon guidance occurs in the absence of neural activity and is considered to be activity-independent. The second process involves glutamatergic excitation, whereby correctly located connections are strengthened by correlated neural activity, whilst removing inappropriately located ones thereby sharpening the topography.The second process is considered to be activitydependent. Here, a number of experiments were undertaken to further examine the interrelationships of activity-dependent and independent processes with respect to functional outcomes. Two models of optic nerve regeneration were studied. In goldfish, following optic nerve crush, regeneration is successful. … In goldfish, guidance along the medio-lateral tectal axis may occur through preordering of axons prior to entering the tectum via the appropriate medial and lateral brachium, with EphA/ephrin-A then guiding axons over the rostral-caudal axis establishing gross topography. The increase in involvement of NMDA-mediated transmission during the period of activity-dependent refinement consolidated the role of this receptor in synapse plasticity. However what triggers NMDA-mediated activity to increase is still largely unresolved, although as the factors governing receptor trafficking during development and synaptic plasticity become better understood, these can be applied to the period of plasticity associated with regenerating axons. And finally, as GABAergic inhibition appears to suppress activity-dependent refinement, means of overcoming this inhibition through 4 such methods as visual training or pharmacological intervention may have significance for mammalian regeneration.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2002. / Härtill 5 uppsatser.