The work in this thesis concerns mechanisms of excitability of neurons. Specifically, it deals with how neurons respond to input, and how their response is controlled by ion channels and other active components of the neuron. I have studied excitability in two systems of the nervous system, the hippocampus which is responsible for memory and spatial navigation, and the peripheral C–fibre which is responsible for sensing and conducting sensory information to the spinal cord. Within the work, I have studied the role of excitability mechanisms in normal function and in pathological conditions. For hippocampus the normal function includes changes in excitability linked to learning and memory. However, it also is intimately linked to pathological increases in excitability observed in epilepsy. In C–fibres, excitability controls sensitivity to responses to stimuli. When this response becomes enhanced, this can lead to pain. I have used computational modelling as a tool for studying hyperexcitability in neurons in the central nervous system in order to address mechanisms of epileptogenesis. Epilepsy is a brain disorder in which a subject has repeated seizures (convulsions) over time. Seizures are characterized by increased and highly synchronized neural activity. Therefore, mechanisms that regulate synchronized neural activity are crucial for the understanding of epileptogenesis. Such mechanisms must differentiate between synchronized and semi synchronized synaptic input. The candidate I propose for such a mechanism is the fast outward current generated by the A-type potassium channel (KA). Additionally, I have studied the propagation of action potentials in peripheral axons, denoted C–fibres. These C–fibres mediate information about harmful peripheral stimuli from limbs and organs to the central nervous system and are thereby linked to pathological pain. If a C–fibre is activated repeatedly, the excitability is altered and the mechanisms for this alteration are unknown. By computational modelling, I have proposed mechanisms which can explain this alteration in excitability. In summary, in my work I have studied roles of particular ion channels in excitability related to functions in the nervous system. Using computational modelling, I have been able to relate specific properties of ion channels to functions of the nervous system such as sensing and learning, and in particular studied the implications of mechanisms of excitability changes in diseases. / <p>QC 20102423</p>

Schädel-Hirn-Traumata sind die häufigste Ursache von Tod und Behinderung bei jungen Erwachsenen und oftmals ein Grund für Erwerbsunfähigkeit. Deshalb sind das Verständnis der zugrundeliegenden Pathomechanismen und die Entwicklung von Rehabilitationsstrategien von höchster Wichtigkeit. Einer der Hauptschädigungsmechanismen sind diffuse axonale Schädigungen. Diese treten insbesondere in frontalen Hirnregionen auf und führen entsprechend zu einer Beeinträchtigung exekutiver Funktionen und Veränderungen im Verhalten noch Jahre nach dem Ereignis. Unsere Studie mit Patienten im chronischen Stadium zielte auf eine genauere Charakterisierung frontaler Funktionen nach Schädel-Hirn-Trauma. In einem ersten behavioralen Experiment setzten wir zwei Paradigmen ein, die einerseits mit dem frontolateralen (Stroop-Interferenz-Aufgabe) und andererseits mit dem frontomedianen Kortex (Aufgabe zur Unterdrückung von Handlungsimitation) assoziiert sind (Schroeter et al., 2007). Die Patienten waren spezifisch in der Aufgabe zur Unterdrückung von Handlungsimitation als Hinweis auf eine Alteration des anterioren frontomedianen Kortex beeinträchtigt. Die Defizite waren hierbei eng mit Veränderungen des Verhaltens und der posttraumatischen Amnesie, die das Outcome nach Schädel-Hirn-Trauma vorhersagt, verbunden. In einem zweiten fMRT-Experiment überprüften wir die Hypothese einer frontomedianen Dysfunktion mittels eines Paradigmas, das spezifisch frontomediane Strukturen beansprucht. Hierbei wurden evaluative Urteile mit semantischen Gedächtnisinhalten kontrastiert („Angela Merkel ist eine gute Bundeskanzlerin“ vs. „Angela Merkel ist Bundeskanzlerin“). Die Ergebnisse bestätigen, dass Patienten nach Schädel-Hirn-Trauma durch persistierende frontomediane Beeinträchtigungen charakterisiert sind. Unsere Ergebnisse stimmen gut mit der Literatur überein, welche Defizite bei der „Theory of Mind“ und sozialen Kognition, die ebenfalls wesentlich mit dem frontomedianen Kortex verbunden sind, berichtet. Die Studie trägt zum Verständnis der Pathomechanismen nach Schädel-Hirn-Trauma bei. Frontomediane Alterationen scheinen wesentlich für die Langzeitfolgen verantwortlich zu sein. Deshalb sollten frontomediane Funktionen in der Diagnostik, insbesondere zur Einschätzung der Prognose, und der Rehabilitation eine stärkere Beachtung finden.

The key to understanding any system, including physiologic and pathologic systems, is to obtain a truly comprehensive view of the system. The purpose of this dissertation was to develop foundational analytical and modeling tools, which would enable such a comprehensive view to be obtained of any physiological or pathological system by combining experimental, clinical, and theoretical viewpoints. Specifically, we focus on the development of analytical and modeling techniques capable of predicting and prioritizing the mechanisms, emergent dynamics, and underlying principles necessary in order to obtain a comprehensive system understanding. Since physiologic systems are inherently complex systems, our approach was to translate the philosophy of complex systems into a set of applied and quantitative methods, which focused on the relationships within the system that result in the system's emergent properties and behavior. The result was a set of developed techniques, referred to as relational modeling and analysis that utilize relationships as either a placeholder or bridging structure from which unknown aspects of the system can be effectively explored. These techniques were subsequently tested via the construction and analysis of models of five very different systems: synaptic neurotransmitter spillover, secondary spinal cord injury, physiological and pathological axonal transport, and amyotrophic lateral sclerosis and to analyze neurophysiological data of in vivo cat spinal motoneurons. Our relationship-based methodologies provide an equivalent means by which the different perspectives can be compared, contrasted, and aggregated into a truly comprehensive viewpoint that can drive research forward.

Os nervos periféricos são extensões do sistema nervoso central e responsável pela interação das atividades entre as extremidades, em suas funções sensitivas e motoras. São vulneráveis aos mesmos tipos de traumas que afetam outros tecidos: contusão, compressão, esmagamento, estiramento, avulsão e laceração. As lesões de nervos periféricos situam-se entre as mais incapacitantes que acometem indivíduos em idade produtiva, em face dos múltiplos aspectos concernentes às sequelas deste tipo de afecção. Desta forma, a interrupção de continuidade da estrutura do nervo, como no caso da neurotmese, por algum tipo de trauma, resulta na interrupção de transmissão dos impulsos nervosos e na desorganização de suas atividades funcionais. Por meio da utilização da microcirurgia foi possível desenvolver técnicas reparadoras que vão desde simples neurorrafia término-terminal até sofisticados procedimentos cirúrgicos com a utilização de enxertos de nervos, veias e artérias invertidas, tubos sintéticos de materiais variados, tais como silicone e polietileno. Outro aspecto que intriga pesquisadores de todo mundo é a utilização de fatores neurogênicos capazes de acelerar ou melhorar a regeneração de nervos periféricos. A gordura autóloga tem sido continuamente referenciada pela sua abundante oferta, no próprio sitio cirúrgico, apresentando resultados promissores, visto que a adventícia dos vasos é constituída por tecido conjuntivo frouxo, rico em adipócitos. Assim, em um trauma, os neuritos oriundos do coto proximal do nervo lesado, ficam diretamente em contato com esses adipócitos. Seguindo este raciocínio, e com base em trabalhos anteriores onde foi usada veia preenchida com músculo esquelético a fresco como enxerto, decidiu-se testar a possibilidade de crescimento axonal por meio de enxerto com tubo de polietileno preenchido por tecido adiposo autólogo associado a protocolo de imersão em câmara hiperbárica, por meio de um estudo Randomizado Controlado. Para tanto utilizou-se um tubo com 12 mm de comprimento por 0,25 mm de diâmetro, com poros de 80 &#x3BC;m de diâmetro, preenchido com tecido adiposo in natura retirado das adjacências do referido nervo, na tentativa de se recuperar o nervo isquiático. Os resultados morfométricos demonstraram que, os grupos experimentais com e sem preenchimento de gordura tiveram resultados, do ponto de vista morfométrico e funcional sem diferenças estatisticamente significantes, contudo, quando estes foram confrontados ao grupo controle final, apresentaram diferenças estatisticamente significantes. Já relevando a avaliação funcional, por meio do Catwalk, constatou-se que não houve diferença estatisticamente significante entre os grupos experimentais, mas teve diferença ao comparar com o grupo controle final, Diante das evidências encontradas e apoiados na literatura pode-se concluir que a câmara hiperbárica trouxe resultados positivos verificados pela aproximação dos resultados dos grupos experimentais tanto morfométrica como funcionalmente. / The peripheral nerves are extensions of the central nervous system and are responsible for the sensory and motor functions of the limbs. These nerves are vulnerable to the same types of traumas that affect other tissues: contusion, compression, crushing, stretching, avulsion, and laceration. Amongst the most disabling kinds of injuries that affect working-age individuals are those of the peripheral nerves; due to the multifaceted characteristics of the aftereffects of the injury. The break in continuity of the nerve structure due to trauma, as in the case of neurotmesis, results in the disruption of the transmission of nerve impulses and the disorganization of their functions. Through the use of microsurgery, it was possible to develop reconstructive techniques that range from a simple end-to-end neurorrhaphy to sophisticated surgical procedures that utilize nerve grafts, inverted veins and arteries, and synthetic rods of varied materials such as silicone or porous polyethylene. Another aspect that intrigues researchers around the world is the utilization of neurogenic factors capable of accelerating or improving the regeneration of peripheral nerves. Autologous fat has been a constant reference in this field of surgery due to its abundant supply at the surgical site itself. The results are promising, as the adventitia of vessels consists of loose connective tissue rich in adipocytes. Thus in a trauma, the neurites derived from the proximal stump of the damaged nerve are in direct contact with these adipocytes. Following this reasoning, and based on previous studies where veins grafted with fresh skeletal muscle were used, we decided to conduct a randomized controlled study to test the possibility of axonal growth by means of grating with a polyethylene rod filled with autologous adipocytes associated with immersion in a hyperbaric chamber. In an attempt to recover the sciatic nerve, a rod 12 mm in length, with a diameter of 0.25 mm, and with pores of 80 &#x3BC;m in diameter, filled with adipose tissue in natura removed from the surroundings of said nerve, was used. The morphometric results showed that the experimental groups with and without fat fillings had results that, from the morphometric and functional point of view, were of no statistically significant difference. However, when these results were compared with the final control group, statistically significant differences were noted. Highlighting the functional evaluation through the use of Catwalk, it was found that there were no statistically significant differences between the experimental groups, but there was indeed a difference in comparison to the final control group. In light of the evidence found and supported by literature, one can conclude that the use of the hyperbaric chamber brought positive results verified by the proximity of both the morphometric and functional results of the experimental groups.

Au cours des dernières années, la synthèse des protéines axonales a été établie comme un mécanisme important pour réguler correctement la réactivité spatiale et temporelle des neurones aux variations de leur microenvironnement, en particulier lors du développement axonal et de la régénération. Pour cela, les transcrits d'ARNm doivent être localisés dans les axones afin d'être traduits. De fait, plusieurs populations d'ARNm ont été identifiées le long des axones de divers types de neurones vertébrés. Le transport approprié des ARNm du corps cellulaire vers le compartiment axonal nécessite des séquences ou des structures spécifiques, généralement trouvées dans le 3'UTR du transcrit. Seules quelques études ont confirmé que le transport et la traduction des ARNm ont lieu dans les axones des vertébrés vivants et que ces mécanismes peuvent être impliqués dans des fonctions neuronales distinctes, comme le maintien de l'homéostasie axonale, le guidage, la croissance et la ramification axonales. Notre laboratoire a précédemment démontré in vivo la présence d'ARNm spécifiques, comme le transcrit de nefma, dans les axones en croissance chez l'embryon de poisson zèbre. En utilisant un système rapporteur développé au sein du laboratoire, il a été démontré que le transport axonal (ou la rétention au corps cellulaire) de plusieurs transcrits dépendait de leur 3'UTR. Se basant sur ces résultats importants, dans une première partie de ce travail, nous avons cherché à étudier la fonction du transcrit nefma transporté dans les axones en développement de l'embryon de poisson zèbre. En effet, Nefma est une protéine cytosquelette propre aux neurones, dont l'expression est déclenchée lors de la différenciation neuronale. Nous avons montré que l’immunoréactivité 3A10 est réduite à mesure que la concentration de MO augmente et que ce marquage est utile pour tester l'efficacité du MO, suggérant que l'anticorps 3A10 pourrait reconnaître nefma. Nous avons également démontré que les neurones de Mauthner se différencient au bon moment et au bon endroit chez les morphants. De plus, nous avons constaté que le « zigzagging » des axones morphants augmente avec la concentration de MO et que la protéine mbp s'accumule inégalement autour des faisceaux axonaux dans les morphants nefma. Cependant, les défauts de perte de fonction de nefma ne sont pas totalement pénétrants et difficiles à quantifier. En outre, dans une deuxième partie de la présente étude, nous avons mis au point une technique de détection de la traduction axonale d'ARNm spécifiques dans le même modèle in vivo. Pour cela, nous avons développé un système inspiré de la technique «TimeSTAMP» développée par l'équipe de Roger Tsien, qui nous permet d'identifier les sites de traduction en étiquetant de manière ingénieuse les protéines nouvellement synthétisées. / In recent years, axonal protein synthesis has been established as an important mechanism to fine regulate spatial and temporal neuronal responsiveness to the varying microenvironment, especially during axonal development and regeneration. For that, mRNA transcripts have to be localized to the axons in order to be translated. In fact, several mRNA populations have been identified along the axons of diverse vertebrate neuronal types. The proper transport from the cell body to the axonal compartment requires specific sequences or mRNA structures, usually found in the 3’UTR of the transcript. Only a few studies have confirmed that mRNA transport and translation take place in axons of living vertebrates, and that these mechanisms can be involved in distinct neuronal functions, as the maintenance of axonal homeostasis, pathfinding, and axonal growth and branching. Our lab previously demonstrated in vivo the presence of specific mRNAs, as nefma transcript, in growing axons of the zebrafish embryo. Thanking advantage of a reporter system developed in the lab, it was shown that axonal transport (or retention at the cell body) of several transcripts depended on their 3’UTR.Building upon these important results, in a first part of this work, we sought to investigate the function of the axonally transported nefma in the developing axons of the zebrafish embryo. Indeed, Nefma is a neuron-specific cytoskeletal protein, which expression is triggered during neuronal differentiation. We showed that the 3A10 signal is reduced as the MO concentration increases and this staining is a useful readout for the efficiency of the MO, suggesting that the 3A10 antibody might recognize nefma. We also demonstrated that the Mauthner neurons differentiate at the right time and place in the morphants. Moreover, we saw that the morphant axons zigzagging increases with increasing MO concentrations and that mbp accumulates in patches around axonal bundles in nefma morphants. However, nefma loss-of-function defects are not totally penetrant and difficult to quantify. Furthermore, in a second part of the present study, we aimed at optimizing a technique facilitating the visualization of axonal translation of specific mRNAs in the same in vivo model. For this, we developed a translation reporter system, inspired on the ‘TimeSTAMP’ technique developed by Roger Tsien’s team, which allows the identification of translation sites along the axons by labeling newly synthesized protein in an ingenious fashion.

Ce travail de thèse vise à mieux décrire les mécanismes de lésions de la tête humaine en situation de choc en optimisant le modèle par éléments finis de la tête humaine de Strasbourg (SUFEHM) en termes de modélisation mécanique du crâne et du cerveau grâce à de nouvelles données expérimentales et de techniques récentes d’imagerie médicales. Une première étape a consisté à améliorer la loi de comportement de la boîte crânienne, valider son comportement en regards d’éléments expérimentaux sur cadavres et proposer un MEF capable de reproduire fidèlement la fracture crânienne. La deuxième partie consiste en la prise en compte pour la première fois de l’anisotropie dans les simulations par EF d’accidents réels en utilisant l’Imagerie du Tenseur de Diffusion. Après implémentation, une phase de validation a été entreprise afin de démontrer l’apport de l’anisotropie de la matière cérébrale dans un MEF. Enfin 125 accidents réels ont été reproduits avec le SUFEHM ainsi amélioré. Une étude statistique sur les paramètres mécaniques calculés a permis de proposer des limites de tolérances en termes de fracture crânienne et de lésions neurologiques en s’intéressant tout particulièrement à l’élongation axonale maximale admissible, nouvelle métrique proposée. / The principal objective of this study is to enhance the existing finite element head model. A composite material model for skull, taking into account damage is implemented in the Strasbourg University Finite Element Head Model in order to enhance the existing skull mechanical constitutive law. The skull behavior is validated in terms of fracture patterns and contact forces by reconstructing 15 experimental cases in collaboration with Medical College of Wisconsin. The new skull model is capable of reproducing skull fracture precisely. The composite skull model is validated not only for maximum forces, but also for lateral impact against actual force time curves from PMHS for the first time. This study also proposes the implementation of fractional anisotropy and axonal fiber orientation from Diffusion Tensor Imaging of 12 healthy patients into an existing human FE head model to develop a more realistic brain model with advanced constitutive laws. Further, the brain behavior was validated in terms of brain strain against experimental data. A reasonable agreement was observed between the simulation and experimental data. Results showed the feasibility of integrating axonal direction information into FE analysis and established the context of computation of axonal elongation in case of head trauma. A total 125 reconstructions were done by using the new advanced FEHM and the axonal strain was found to be the pertinent parameter to predict DAI.

Les connexions neuronales du système visuel forment des synapses spatialement distribuées en couches discrètes. Comprendre la base du ciblage spécifique axonale est critique pour déchiffrer la formation des réseaux neuronaux complexes. Dans une première étude, nous avons investigué le rôle de la protéine de la matrice extracellulaire Reelin dans la formation in vivo du circuit rétinotectal chez le poisson zèbre. Ce circuit se compose de cellules ganglionnaires de la rétine (CGRs) transmettant l’information visuelle au cerveau via la projection de leur axone dans les différentes couches du tectum optique. Nous avons démontré que la Reelin secrétée par de neurones inhibiteurs localisés dans les couches supérieures du tectum optique forme un gradient. L’induction de mutations délétères dans la voie de signalisation canonicale de la Reelin à l’aide d’outils génétiques a conduit à des défauts de ciblage des axones de CGRs. Nos résultats démontrent un nouveau rôle de la Reelin lors du développement du système visuel et la décrivent comme signature moléculaire nécessaire au ciblage et au positionnement précis des axones de CGRs.Dans une seconde étude, nous avons utilisé la technique CRISPR/Cas9 pour développer une nouvelle approche de mutagénèse conditionnelle chez le poisson zèbre. Nos résultats démontrent que la perturbation de gènes dans des tissues spécifiques peut être effectué par l’induction de l’expression de la protéine Cas9 via le système Gal4/UAS. Nous avons établis un outil pour induire l’apparition de mutations délétères dans des clones de cellules mais aussi dans des cellules individuelles, tous pouvant être suivit distinctement grâce à un marquage génétique. / Neuronal connections in the visual system are arranged in synaptic laminae. Understanding the basis of lamina-specific axonal targeting is critical to gain deeper insights on how complex neural networks form. In a first study we investigated the role of the ECM protein Reelin during zebrafish retinotectal circuit formation in vivo. Here retinal ganglion cells (RGCs) convey the visual information to the brain by projecting their axons to different layers of the optic tectum. We demonstrated that Reelin secreted by a specific class of tectal superficial inhibitory neurons is spatially distributed in a superficial-to-deep gradient within the tectal neuropil. Induced gene disruption for all the components of the canonical Reelin pathway expressed in the retinotectal system resulted in aberrant layering of RGC axons suggesting a role for Reelin pathway in axonal sublaminar segregation. Altogether our findings elucidate a new role for Reelin in vertebrate visual system development, during which it acts as molecular cue by imparting positional information for ingrowing RGCs.In a second study we took advantage of the CRISPR/Cas9 technology to develop a novel approach for conditional mutagenesis in zebrafish. Our results provide evidence that tissue-specific gene disruption can be achieved by driving Cas9 expression with the Gal4/UAS system. We established a tool to induce loss-of-function mutations in cell clones or single cells that can be followed by genetic labeling, enabling their phenotypic analysis. Our technique has the potential to be applied to a wide-range of model organisms, allowing systematic mutagenesis and labeling on a genome-wide scale.

La régénération après une lésion de la moelle épinière (ME) est abortive. Elle est du à une cascade d'événements cellulaires et moléculaires, dont la rupture de la barrière hémato-encéphalique, une inflammation persistante, une cicatrice gliale et la formation d'une cavité, combinée à la présence de molécules inhibitrices pour la repousse. Actuellement, aucune thérapie n'est efficace, mais le design de biomatériaux implantables pourrait permettre leur développement. L'hydrogel de chitosane (hCh) apparait prometteur, notamment grâce à la modulation de ses propriétés biologiques, notamment en modifiant son degré de déacétylation (DA). J'ai donc testé, dans une hémisection dorsale de la ME de rat, différentes formulations d'hCh fragmenté et examiné leur capacité à s'intégrer dans le tissu hôte lésé, sans produire une inflammation ou une réaction astrocytaire excessive. Mon travail montre que l'implantation de fragments d'hCh avec un DA de 4% est favorable à la reconstruction du tissu, en attirant différents types cellulaires et en recréant une vascularisation fonctionnelle. Il permet aussi de moduler la réponse inflammatoire, en favorisant une polarisation des macrophages en un phénotype M2. La cicatrice gliale est aussi réduite et les processus astroytaires s'orientent vers l'implant, en s'associant aux axones qui régénèrent et caractérisés par traçage axonale et immunohistochimie. Plusieurs sont myélinisés ou entourés par des cellules de Schwann, au moins jusqu'à 10 semaines après la lésion. Enfin, le remodelage structural est associé à une récupération locomotrice significative. / Regeneration after traumatic spinal cord injury generally fails due to a cascade of cellular and molecular events, including blood-spinal cord barrier breakdown,persistent and uncontrolled inflammation, and glial scarring and cavity formation combined with the presence of axon growth-inhibitory molecules. While efficient therapies are still lacking, recent progress in the design of implantable biomaterials may well open up new possibilites for their development. Chitosan hydrogels (hCh) seem particularly promising as their biological properties can be fine-tuned, notably by their degree of acetylation (DA). In the context of a rat dorsal spinal cord hemisection, I have tested different formulations of fragmented hCh for their ability to integrate into lesioned host tissue without creating additional inflammation, or excessive astrocytic reaction. Thus, I found that implantation of hCh particles of 4% DA allows for tissue reconstruction by attracting different cell types and recreating a functional vasculature. Importantly, it modulates the inflammatory response, favoring polarization of invading macrophages towards the M2 phenotype. In lesioned-implanted animals, the glial scar is less fibrous, astrocyte processes are mainly oriented towards the lesion and accompany a robust regrowth of fibers, whose origin was identified by axon tracing and immunohistochemistry. Many of these fibers are myelinated or ensheathed by Schwann cells, maintained at long term in the implant. Finally, this structural remodeling is associated with significant, long-lasting recovery of locomotor function, as I have shown by open-field and gait analysis.

L’émergence de la notion selon laquelle la réponse inflammatoire exercerait des effets bénéfiques dans la pathologie ischémique cérébrale, en particulier au cours de la phase de récupération fonctionnelle nous a conduit à étudier l’implication des cellules microgliales dans le déclenchement des mécanismes de neuroplasticité post-ischémique. Notre étude a été réalisée chez le Rat soumis à une ischémie focale permanente induite par photothrombose. L’activation microgliale a été modulée par un traitement au 3-aminobenzamide (3-AB), un inhibiteur spécifique de la poly(ADP-ribose)polymérase-1, jouant un rôle prépondérant dans l’activation de ces cellules. Nos données montrent que le 3-AB entraîne une diminution importante de l’activation microgliale aux temps courts associée à plus long terme à une réduction de l’expression de la synaptophysine et de GAP-43, respectivement marqueurs des processus de synaptogenèse et croissance axonale. L’ensemble de ces données indique donc que les cellules microgliales constituent effectivement des acteurs cellulaires essentiels de la neuroplasticité post-ischémique. Le Brain-derived neurotrophic factor (BDNF) se révélant un candidat potentiellement capable de promouvoir de tels changements, nous avons pu mettre en évidence que ces cellules représentaient de façon précoce une source importante de BDNF. Ces résultats ont été confirmés par la nette diminution des taux de BDNF mesurés dans les zones corticales lésionnelles et péri-lésionnelles des animaux traités par le 3-AB. Dans un deuxième temps, le métabolisme complexe de cette neurotrophine à travers l’existence de deux formes, pro- et mature, aux effets biologiques opposés, nous a conduit à réaliser une étude spatio-temporelle des expressions post-ischémiques du BDNF total (ELISA), pro- et mature (Western blotting). Aux temps courts (4-24 h), les expressions du BDNF total, pro- et mature sont augmentées dans les territoires corticaux lésés, péri-lésionnels et homotopiques tandis qu’aux temps longs (8-30 j), le BDNF total reste accru dans les régions distantes de la zone infarcie (hippocampes et cortex contralatéral). Concernant les expressions des formes pro- et mature, nos résultats indiquent une augmentation entre 8 et 30 j uniquement dans les territoires hippocampiques. D’un point de vue cellulaire, le BDNF est exprimé du côté ipsilatéral dans les neurones et les cellules non neuronales tandis que du côté contralatéral, l’expression est limitée aux neurones. Nos résultats tout en faisant apparaître des divergences importantes dans les variations d’expressions du BDNF total (ELISA) et des différentes formes (Western blotting) indiquent que la mesure du BDNF total doit être couplée à une étude permettant de discriminer les deux formes. De plus, tout en confirmant l’implication de cette neurotrophine dans les mécanismes adaptatifs induits en réponse à une ischémie cérébrale, ces données suggèrent que les territoires distants de la zone lésée jouent un rôle majeur dans ces processus. / Evidences showing that under certain circumstances, inflammatory response could be neuroprotective and could also promote adult neurogenesis are growing. In this context, the objective of this work was to investigate the impact of microglial cells in the neuroplastic events. Rats were subjected to photothrombotic ischemia and microglial cells activation was blocked by the mean of poly(ADP-ribose)polymérase-1 (PARP-1) inhibition using 3- aminobenzamide (3-AB) since this protein has been shown to play a major role in this activation. Our results show that PARP-1 activity reduction was associated with a strong repression of the acute microglial activation. Beside, 3-AB treated animals exhibited a decrease in synaptophysin (synaptogenesis) and GAP-43 (axonal growth) expressions. Taken together, our data argue for a supportive role of microglial in adaptive brain plasticity events. According to the preponderant contribution of BDNF in these events, assessment of its cellular localization was performed, and confirmed that these cells represent a significant source. Beside, BDNF immunoreactivity (IR) in microglial cells and BDNF levels in the lesioned and surrounding lesioned areas were found decreased in 3-AB treated animals. However, since this neurotrophin can exert ambivalent biological actions through pro- versus mature forms, we investigate the proper effect of cerebral ischemia on total (Elisa), pro- and mature (Western blotting) expressions. Our results show that total, pro- and mature BDNF expressions are augmented in the early times (4-24h) of ischemia within the lesioned, the surrounding non lesioned and the contralateral cortical areas. At longer time points, total BDNF was still increased at 8d in regions distant from the lesion (hippocampi and contralateral cortex) while pro- and mature forms rise between 8d to 30d in hippocampic territories only. In term of cellular distribution, BDNF-IR was found in neurons but also in non neuronal cells ipsilaterally whereas in the opposite side BDNF staining was restricted to neurons. Our data while raising the question of the pertinence of total BDNF expression in a context of studying its supportive potential action indicate that such assessment has to be coupled with the discrimination of both forms. In addition, our data confirm the important role of BDNF in post-stroke adaptive mechanisms and argue in favour of an important contribution of the hippocampal territory and of the contralateral hemisphere in BDNF related post-stroke neuronal circuit remodelling suggesting that strategies targeting this hemisphere are likely to mediate functional compensation.

Traumatic brain injury is a topic that in recent years has received increased scrutiny by the media and is viewed as a cause for public health concern in athletes that are participating in contact sports. There has been an apparent rise in the reported number of traumatic brain injuries over the last decade possibly due to a number of factors such as an increase in enrollment of sports and suspected better understanding of brain injury in the sports world. Direct or indirect impact forces applied involving acceleration/deceleration and linear/angular forces primarily cause trauma to the brain. This insult results in evident diffuse axonal and focal injuries to varying degrees in brain tissue. The spectrum of pathophysiology in traumatic brain injury involves structural, neurochemical, metabolic, vascular, inflammatory, immunologic, and ultimately cell death, which plays a hand directly in the nonspecific presentation of symptoms reported by athletes as well as the progression of recovery. Traumatic brain injury is typically associated with short- and long-term sequelae, however, inducing repetitive episodes of trauma over a career, as may happen in sports, may lead to a progressive neurodegenerative disease known as chronic traumatic encephalopathy. Chronic traumatic encephalopathy has been known to affect boxers previously, but in recent years the attention has shifted and found this disease in athletes from other sports. The spectrum of disease in chronic traumatic encephalopathy involves a progressive tauopathy that spreads across different regions of the brain in a classified four staged grading system. Several risk factors have been identified in placing athletes at risk for traumatic brain episodes, however no risk factors have been directly linked to chronic traumatic encephalopathy. Much information is lacking in a complete understanding of traumatic brain injury and chronic traumatic encephalopathy, therefore emphasizing the importance of further research and consistently improving modifications in the protocols for assessment, recognition, management, and return to play criteria for injured athletes. Furthermore, despite the gaps in knowledge, preventative measures should serve a particular role in reducing the incidence of detected traumatic brain injuries, which should include policy changes, sport rule changes, and especially changes to the accepted sports culture through mandatory education.