La neurogenèse persiste à l’âge adulte dans deux régions du système nerveux central (SNC) des mammifères : la zone sous-ventriculaire (SVZ) du cerveau antérieur et la zone sous-granulaire (SGZ) de l’hippocampe. Cette neurogenèse est possible grâce à la capacité de prolifération des cellules souches présentes dans les niches de la SVZ et la SGZ, mais en vieillissant, le cerveau subit une diminution dramatique du nombre de cellules souches neurales adultes (CSNa), une diminution de la prolifération cellulaire et une altération des niches de neurogenèse. Cependant, une importante question reste sans réponse : comment la perte tardive des CSNa est temporellement reliée aux changements de l’activité de prolifération et de la structure de la principale niche de neurogenèse (la SVZ)? Afin d’avoir un aperçu sur les événements initiaux, nous avons examiné les changements des CSNa et de leur niche dans la SVZ entre le jeune âge et l’âge moyen. La niche de la SVZ des souris d’âge moyen (12 mois) subit une réduction de l’expression des marqueurs de plusieurs sous-populations de précurseurs neuraux en comparaison avec les souris jeunes adultes (2 mois). Anatomiquement, cela est associé avec des anomalies cytologiques, incluant une atrophie générale de la SVZ, une perte de la couche de cellules sousépendymaires par endroit et l’accumulation de gouttelettes lipidiques de grande taille dans l’épendyme. Fonctionnellement, ces changements sont corrélés avec une diminution de l’activité de la SVZ et une réduction du nombre de nouveaux neurones arrivant aux bulbes olfactifs. Pour déterminer si les CSNa de la SVZ ont subi des changements visibles, nous avons évalué les paramètres clés des CSNa in vivo et in vitro. La culture cellulaire montre qu’un nombre équivalent de CSNa ayant la capacité de former des neurosphères peut être isolé du cerveau du jeune adulte et d’âge moyen. Cependant, à l’âge moyen, les précurseurs neuraux semblent moins sensibles aux facteurs de croissance durant leur différenciation in vitro. Les CSNa donnent des signes de latence in vivo puisque leur capacité d’incorporation et de rétention du BrdU diminue. Ensemble, ces données démontrent que, tôt dans le processus du vieillissement, les CSNa et leur niche dans la SVZ subissent des changements significatifs, et suggèrent que la perte de CSNa liée au vieillissement est secondaire à ces événements. / Neurogenesis persists throughout the adulthood in two regions of the mammalian central nervous system (SNC): the sub-ventricular zone (SVZ) of the forebrain and the sub-granular zone (SGZ) of the hippocampus. Neurogenesis is possible due to the proliferation capacity of stem cells present within both the SVZ and SGZ niches, but with aging, the forebrain undergoes a drastic reduction in its number of adult neural stem cells (aNSCs), a decrease of cell proliferation and an alteration of the neurogenic niches. However, a key unresolved question remains: how the onset of aNSC loss is temporally related to changes of proliferating activity and to structural alterations within the principal stem cell niche (the SVZ)? To gain insights into the initial events leading to aging-associated aNSC loss, we investigated the changes occurring to aNSCs and the SVZ niche between young adulthood and middle-age. The SVZ niche of middle-aged mice (12-months-old) was found to display reduced expression of markers for multiple neural precursor sub-populations when compared to young adult mice (2-months-old). Anatomically, this was associated with significant cytological aberrations, including an overall atrophy of the SVZ, loss of sub-ependymal cells, and accumulation of large lipid droplets within the ependyma. Functionally, these changes correlated with diminished SVZ activity and reduced number of newly born neurons reaching the principal target tissue: the olfactory bulbs. To determine whether changes were evident at the level of the SVZ stem cells, we evaluated key in vitro and in vivo parameters of aNSCs. Tissue culture experiments showed that equal numbers of neurosphere-forming aNSCs could be isolated from young adult and middle-aged forebrains. However, at middle-age, neural precursors seemed to be less sensitive to growth factors during their in vitro differentiation and displayed signs of increased quiescence in vivo. Collectively, these findings demonstrate that, with early aging, aNCS and their SVZ niche go through significant changes, and suggest that aging-associated aNSC loss is secondary to these events.

Background: Amyotrophic lateral sclerosis (ALS) is a late-onset progressive neurodegenerative disorder. ALS has been traditionally believed to be primarily a motor neuron disease. However, accumulating data indicate that loss of contact between the axons and the muscle fibres occurs early; long before the death of motor neurons and that muscle fibres may initiate motor neuron degeneration. Thus, the view of ALS is changing focus from motor neurons alone to also include the muscle fibres and the neuromuscular junctions (NMJs). While skeletal muscles are affected in ALS, oculomotor disturbances are not dominant features of this disease and extraocular muscles (EOMs) are far less affected than limb muscles. Why oculomotor neurons and EOMs are capable to be more resistant in the pathogenetic process of ALS is still unknown. The overall goal of this thesis is to explore the pathophysiology of ALS from a muscle perspective and in particular study the expression and distribution of key neurotrophic factors (NTFs) and Wnt proteins in EOMs and limb muscles from ALS donors and from SOD1G93A transgenic mice. Comparisons were made with age-matched controls to distinguish between changes related to ALS and to ageing. Results: Brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4) were present in EOMs and limb muscles at both mRNA and protein levels in control mice. The mRNA levels of BDNF, NT-3 and NT-4 were significantly lower in EOMs than in limb muscles of early and/or late control mice, indicating an intrinsic difference in NTFs expression between EOMs and limb muscles. qRT-PCR analysis showed significantly upregulated mRNA levels of NT-3 and GDNF in EOMs but significantly downregulated mRNA levels of NT-4 in limb muscles from SOD1G93A transgenic mice at early stage. The NTFs were detected immunohistochemically in NMJs, nerve axons and muscle fibres. The expression of BDNF, GDNF and NT-4 on NMJs of limb muscles, but not of EOMs, was significantly decreased in terminal stage ALS animals as compared to the limb muscles of the age-matched controls. In contrast, NTFs expression in intramuscular nerve axons did not present significant changes in either muscle group of early or late ALS mice. NTFs, especially BDNF and NT-4 were upregulated in some small-sized muscle fibres in limb muscles of late stage ALS mice. All the four Wnt isoforms, Wnt1, Wnt3a, Wnt5a and Wnt7a were detected in most axon profiles in all human EOMs with ALS, whereas significantly fewer axon profiles were positive in the human limb muscles except for Wnt5a. Similar differential patterns were found in myofibres, except for Wnt7a, where its expression was elevated within sarcolemma of limb muscle fibres. β-catenin, a marker of the canonical Wnt pathway was activated in a subset of myofibres in the EOMs and limb muscle in all ALS patients. In the SOD1G93A mouse, all four Wnt isoforms were significantly decreased in the NMJs at the terminal stage compared to age matched controls. Conclusions: There were clear differences in NTF and Wnt expression patterns between EOM and limb muscle, suggesting that they may play a role in the distinct susceptibility of these two muscle groups to ALS. In particular, the early upregulation of GDNF and NT-3 in the EOMs might play a role in the preservation of the EOMs in ALS. Further studies are needed to determine whether these proteins and the pathways they control may be have a future potential as protecting agents for other muscles.

La sclérose latérale amyotrophique (SLA) est la maladie des neurones moteurs la plus fréquente, affectant 4-6 individus par 100,000 habitants à l’échelle mondiale. La maladie se caractérise par une faiblesse et une atrophie musculaire suite à la dégénérescence des neurones du cortex moteur, tronc cérébral et moelle épinière. Les personnes atteintes développent les premiers symptômes à l’âge adulte et la maladie progresse sur une période de trois à cinq ans. Il a été répertorié qu’environ 10% des patients ont une histoire familiale de SLA; 90% des gens affectés le sont donc de façon sporadique. La découverte il y a 19 ans de mutations dans le gène zinc/copper superoxide dismutase (SOD1), présentes dans 15-20% des cas familiaux de SLA et environ 2% du total des individus affectés, a été l’événement déclencheur pour la découverte de variations génétiques responsables de la maladie. La recherche sur la génétique de la SLA a connu une progression rapide ces quatre dernières années avec l’identification de mutations dans de nouveaux gènes. Toutefois, même si certains de ces gènes ont été démontrés comme réellement liés à la maladie, la contribution d’autres gènes demeure incertaine puisque les résultats publiés de ceux-ci n’ont pas, à ce jour, été répliqués. Une portion substantielle de cas reste cependant à être génétiquement expliquée, et aucun traitement à ce jour n’a été démontré comme étant efficace pour remédier, atténuer ou prévenir la maladie. Le but du projet de recherche de doctorat était d’identifier de nouveaux gènes mutés dans la SLA, tout en évaluant la contribution de gènes nouvellement identifiés chez une importante cohorte multiethnique de cas familiaux et sporadiques. Les résultats présentés sont organisés en trois sections différentes. Dans un premier temps, la contribution de mutations présentes dans le gène FUS est évaluée chez les patients familiaux, sporadiques et juvéniles de SLA. Précisément, de nouvelles mutations sont rapportées et la proportion de mutations retrouvées chez les cas familiaux et sporadiques de SLA est évaluée. De plus, une nouvelle mutation est rapportée dans un cas juvénile de SLA; cette étude de cas est discutée. Dans un deuxième temps, de nouvelles avenues génétiques sont explorées concernant le gène SOD1. En effet, une nouvelle mutation complexe est rapportée chez une famille française de SLA. De plus, la possibilité qu’une mutation présente dans un autre gène impliqué dans la SLA ait un impact sur l’épissage du gène SOD1 est évaluée. Finalement, la dernière section explique la contribution de nouveaux gènes candidats chez les patients atteints de SLA. Spécifiquement, le rôle des gènes OPTN, SIGMAR1 et SORT1 dans le phénotype de SLA est évalué. Il est souhaité que nos résultats combinés avec les récents développements en génétique et biologie moléculaire permettent une meilleure compréhension du mécanisme pathologique responsable de cette terrible maladie tout en guidant le déploiement de thérapies suite à l’identification des cibles appropriées. / Amyotrophic lateral sclerosis (ALS) is the most common of motor neuron diseases, affecting 4-6 individuals per 100,000 individuals worldwide. ALS is characterized by muscle weakness and atrophy caused by the degeneration of neurons located in the motor cortex, brain stem and spinal cord. This fatal disease generally has an adult onset and progresses over a three to five year period. While 10% of patients affected have a family history of the disease, 90% of cases do not and are considered sporadic. The finding of mutations in the zinc/copper superoxide dismutase gene (SOD1) gene 19 years ago in about 15-20% of familial ALS (FALS) patients and approximately 2% of overall cases developed the interest of identifying rare genetics variants causing the disease. The ALS research field experienced a rapid progression during the last four years as mutations in new genes have been identified. While mutations in some of those new genes have been clearly linked to ALS, the role of others is still questionable and so far has not been positively replicated in other populations. Importantly, a significant portion of cases still need to be genetically explained and, unfortunately, there is still no effective treatment to cure, attenuate or prevent the disease. The aim of this Ph.D research project was to identify new ALS mutated genes while analysing the causative role of other newly identified genes in a large familial and sporadic ALS cohort of different origins. The results presented here are categorized into three different sections. First, the contribution of FUS mutations to familial, sporadic and juvenile ALS is analysed. Specifically, new FUS mutations are reported in ALS cases and the proportions of variants present in the tested familial and sporadic ALS cohorts are assessed. In addition, a new mutation is reported in a juvenile ALS patient, and this interesting case is discussed. Second, new genetic avenues are explored for the SOD1 gene. Precisely, a new and complex SOD1 mutation is reported in a French ALS family. Moreover, the possibility that other ALS mutated genes influence SOD1 splicing events is evaluated. Third, the contribution of new candidate genes is evaluated. Precisely, the contribution of OPTN, SIGMAR1 and SORT1 genes to the ALS phenotype is assessed. Hopefully, our different findings combined with recent developments in genetics and molecular biology will permit a better understanding of the pathological mechanisms involved in the disease and will lead to the identification of the right targets in order to develop appropriate therapeutics for ALS patients.

No description available.

O presente texto descreve métodos e apresenta resultados do projeto de pesquisa de mestrado intitulado \"Dois Problemas em Análise de Formas de Estruturas de Ramificação\". Ambos os problemas abordados estão relacionados às sub-áreas da Análise de Formas denominadas Caracterização e Descrição de Formas. O primeiro problema consiste na investigação de um conjunto de características propostas para distingüir, primeiramente, entre estruturas de ramificação de vasos sangüíneos em imagens de retina segmentadas manualmente e automaticamente. A seguir, as mesmas características são aplicadas para discernir entre estruturas de ramificação de vasos sangüíneos em imagens de retina com e sem retinopatia diabética proliferativa (Proliferative Diabetic Retinopathy - PDR). A PDR é uma das patologias associadas à diabetes, que pode culminar na cegueira do indivíduo. Diagnósticos são possíveis por meio de imagens de fundo de olho e, quando efetuados precocemente, viabilizam intervenções oportunas evitando a perda da visão. Neste trabalho, 27 imagens digitais de fundo de olho foram segmentadas por dois processos distintos, isto é, segmentação manual por um especialista e a segmentação automática, mediante a transformada contínua Wavelet - CWT e classificadores estatísticos. Visando à caracterização destas formas, um conjunto de 08 características foi proposto. Este conjunto foi formado por três grupos, a saber: descritores tradicionais geométricos (Área, Perímetro e Circularidade), descritores associados à transformada wavelet ( 2o momento estatístico da distribuição de módulos da CWT, Entropia de Orientação da distribuição de fases da CWT e Curvatura) e um descritor fractal (Dimensão de Correlação - Global e Mediana). Uma Análise Discriminante Linear LDA revelou que as características geométricas tradicionais não detectam o início da retinopatia diabética proliferativa. A maior capacidade discriminante individual foi exibida pela Curvatura, com Área sob a curva ROC de 0.76. Um subconjunto com 6 características apresentou grande capacidade discriminante com Área sob a curva ROC de 0.90. O segundo problema diz respeito à extração de contorno de estruturas de ramificação bidimensionais de neurônios tridimensionais. Este trabalho contribui originalmente com uma solução para este problema, propondo dois algoritmos desenvolvidos para Rastreamento de Ramos e Extração do Contorno Paramétrico de estruturas de ramificação, capazes de transpor regiões críticas formadas por cruzamentos ocasionados pela projeção de estruturas 3D no plano das imagens 2D. Grande parte dos métodos baseados em contorno para análise de formas de estruturas de ramificação de células neuronais não produz representações corretas destas formas, devido à presença de sobreposições entre processos neuronais, levando os algoritmos tradicionais de extração de contorno a ignorar as regiões mais internas destas estruturas, gerando representações incompletas. O sistema proposto neste trabalho foi desenvolvido objetivando a solução do problema de extração de contorno, mesmo na presença de múltiplas sobreposições. Inicialmente, a imagem de entrada é pré-processada, gerando um esqueleto 8-conexo com ramos de um pixel de largura, um conjunto de sementes de sub-árvores dendríticas e um conjunto de regiões críticas (bifurcações e cruzamentos). Para cada sub-árvore, o algoritmo de rastreamento rotula todos os pixels válidos de um ramo, até chegar em uma região crítica, onde o algoritmo decide a direção em que deve continuar o rastreamento. Nosso algoritmo mostrou-se robusto, mesmo quando aplicado a imagens com segmentos paralelos muito próximos. Resultados obtidos com imagens reais (neurônios) são apresentados. / This document describes methods and presents results from the Master of Science\'s research project in computer science entitled \"Two Problems in Shape Analysis of Branching Structures\". Both tackled problems herein are related to Shape Analysis sub-fields, namely Characterization and Description of shapes. The former problem consists of an investigation on a proposed set of features aimed at discriminating, firstly, between blood vessels branching structures manually and automatically segmented. In the sequel, the same features are used to assess their discriminative capability in distinguishing between blood vessels branching structures with and withoud proliferative diabetic retinopathy (PDR). The PDR is a pathology related to diabetes, which may lead to the blindness. Diagnosis is possible through optic fundus image analysis, which may allow timely interventions preventing vision loss. In this work, 27 digital optic fundus images were segmented by two distinct segmentation processes, i.e. manual segmentation carried out by an especialist and automated segmentation, through the CWT (Continuous Wavelet Transform) and statistical classifiers. In order to characterize such a shapes, a set of 8 features has been proposed. The aforementioned set was comprised of three features groups, that is: traditional geometric descriptors (Area, Perimeter and Circularity), wavelet-based descriptors (2nd statistical moment from the CWT Modulus distribution, Orientation Entropy from the CWT Phase distribution and Curvature) and a fractal descriptor (Correlation Dimension - global and median). Linear Discriminant Analysis LDA revelead that the traditional geometric features are not able to detect early proliferative diabetic retinopathy. The largest singular discriminant capability was shown by the Curvature, with area under the ROC curve of 0.76. A subset of 6 features presented a good discriminating power with area under the curve of 0.90. The second problem concerns contour extraction from 2D branching structures of 3D neurons. This work contributes with an original solution for such a problem, proposing two algorithms devised for Branches Tracking and Branching Structures Contour Extraction. The proposed algorithms are able to traverse critical regions implied by the projection of 3D structures onto a 2D image plane. Most of contour-based methods intended to shape analysis of neuronal branching structures fall short of yielding proper shape representations, owing to the presence of overlapings among neuronal processes, causing the traditional algorithms for contour following to ignore the innermost regions, thus generating incomplete representations. The proposed framework system was developed aiming at the solution of the contour extraction problem, even in the presence of multiple overlapings. The input image is pre-processed, so as to obtain an 8-connected skeleton with one-pixel wide branches, a set of seeds of dendritic sub-trees and a set of critical regions (bifurcations, crossings and superpositions). For each sub-tree, the Branches Tracking Algorithm labels all valid pixels of a branch, until reaching a critical region, where the algorithm decides about the direction to go on with the tracking. Our algorithm has shown robustness, even in images plenty of very close parallel segments. Results with real images (neurons) are presented.

No description available.

Dans cette thèse, nous abordons le contrôle moteur du mouvement du coude à travers deux approches expérimentales : une première étude psychophysique a été effectuée chez les sujets humains, et une seconde implique des enregistrements neurophysiologiques chez le singe. Nous avons recensé plusieurs aspects non résolus jusqu’à présent dans l’apprentissage moteur, particulièrement concernant l’interférence survenant lors de l’adaptation à deux ou plusieurs champs de force anti-corrélés. Nous avons conçu un paradigme où des stimuli de couleur aident les sujets à prédire la nature du champ de force externe actuel avant qu’ils ne l’expérimentent physiquement durant des mouvements d’atteinte. Ces connaissances contextuelles faciliteraient l’adaptation à des champs de forces en diminuant l’interférence. Selon le modèle computationnel de l’apprentissage moteur MOSAIC (MOdular Selection And Identification model for Control), les stimuli de couleur aident les sujets à former « un modèle interne » de chaque champ de forces, à s’en rappeler et à faire la transition entre deux champs de force différents, sans interférence. Dans l’expérience psychophysique, quatre groupes de sujets humains ont exécuté des mouvements de flexion/extension du coude contre deux champs de forces. Chaque force visqueuse était associée à une couleur de l’écran de l’ordinateur et les deux forces étaient anti-corrélées : une force résistante (Vr) a été associée à la couleur rouge de l’écran et l’autre, assistante (Va), à la couleur verte de l’écran. Les deux premiers groupes de sujets étaient des groupes témoins : la couleur de l’écran changeait à chaque bloc de 4 essais, tandis que le champ de force ne changeait pas. Les sujets du groupe témoin Va ne rencontraient que la force assistante Va et les sujets du groupe témoin Vr performaient leurs mouvements uniquement contre une force résistante Vr. Ainsi, dans ces deux groupes témoins, les stimuli de couleur n’étaient pas pertinents pour adapter le mouvement et les sujets ne s’adaptaient qu’à une seule force (Va ou Vr). Dans les deux groupes expérimentaux, cependant, les sujets expérimentaient deux champs de forces différents dans les différents blocs d’essais (4 par bloc), associés à ces couleurs. Dans le premier groupe expérimental (groupe « indice certain », IC), la relation entre le champ de force et le stimulus (couleur de l’écran) était constante. La couleur rouge signalait toujours la force Vr tandis que la force Va était signalée par la couleur verte. L’adaptation aux deux forces anti-corrélées pour le groupe IC s’est avérée significative au cours des 10 jours d’entraînement et leurs mouvements étaient presque aussi bien ajustés que ceux des deux groupes témoins qui n’avaient expérimenté qu’une seule des deux forces. De plus, les sujets du groupe IC ont rapidement démontré des changements adaptatifs prédictifs dans leurs sorties motrices à chaque changement de couleur de l’écran, et ceci même durant leur première journée d’entraînement. Ceci démontre qu’ils pouvaient utiliser les stimuli de couleur afin de se rappeler de la commande motrice adéquate. Dans le deuxième groupe expérimental, la couleur de l’écran changeait régulièrement de vert à rouge à chaque transition de blocs d’essais, mais le changement des champs de forces était randomisé par rapport aux changements de couleur (groupe « indice-incertain », II). Ces sujets ont pris plus de temps à s’adapter aux champs de forces que les 3 autres groupes et ne pouvaient pas utiliser les stimuli de couleurs, qui n’étaient pas fiables puisque non systématiquement reliés aux champs de forces, pour faire des changements prédictifs dans leurs sorties motrices. Toutefois, tous les sujets de ce groupe ont développé une stratégie ingénieuse leur permettant d’émettre une réponse motrice « par défaut » afin de palper ou de sentir le type de la force qu’ils allaient rencontrer dans le premier essai de chaque bloc, à chaque changement de couleur. En effet, ils utilisaient la rétroaction proprioceptive liée à la nature du champ de force afin de prédire la sortie motrice appropriée pour les essais qui suivent, jusqu’au prochain changement de couleur d’écran qui signifiait la possibilité de changement de force. Cette stratégie était efficace puisque la force demeurait la même dans chaque bloc, pendant lequel la couleur de l’écran restait inchangée. Cette étude a démontré que les sujets du groupe II étaient capables d’utiliser les stimuli de couleur pour extraire des informations implicites et explicites nécessaires à la réalisation des mouvements, et qu’ils pouvaient utiliser ces informations pour diminuer l’interférence lors de l’adaptation aux forces anti-corrélées. Les résultats de cette première étude nous ont encouragés à étudier les mécanismes permettant aux sujets de se rappeler d’habiletés motrices multiples jumelées à des stimuli contextuels de couleur. Dans le cadre de notre deuxième étude, nos expériences ont été effectuées au niveau neuronal chez le singe. Notre but était alors d’élucider à quel point les neurones du cortex moteur primaire (M1) peuvent contribuer à la compensation d’un large éventail de différentes forces externes durant un mouvement de flexion/extension du coude. Par cette étude, nous avons testé l’hypothèse liée au modèle MOSAIC, selon laquelle il existe plusieurs modules contrôleurs dans le cervelet qui peuvent prédire chaque contexte et produire un signal de sortie motrice approprié pour un nombre restreint de conditions. Selon ce modèle, les neurones de M1 recevraient des entrées de la part de plusieurs contrôleurs cérébelleux spécialisés et montreraient ensuite une modulation appropriée de la réponse pour une large variété de conditions. Nous avons entraîné deux singes à adapter leurs mouvements de flexion/extension du coude dans le cadre de 5 champs de force différents : un champ nul ne présentant aucune perturbation, deux forces visqueuses anti-corrélées (assistante et résistante) qui dépendaient de la vitesse du mouvement et qui ressemblaient à celles utilisées dans notre étude psychophysique chez l’homme, une force élastique résistante qui dépendait de la position de l’articulation du coude et, finalement, un champ viscoélastique comportant une sommation linéaire de la force élastique et de la force visqueuse. Chaque champ de force était couplé à une couleur d’écran de l’ordinateur, donc nous avions un total de 5 couleurs différentes associées chacune à un champ de force (relation fixe). Les singes étaient bien adaptés aux 5 conditions de champs de forces et utilisaient les stimuli contextuels de couleur pour se rappeler de la sortie motrice appropriée au contexte de forces associé à chaque couleur, prédisant ainsi leur sortie motrice avant de sentir les effets du champ de force. Les enregistrements d’EMG ont permis d’éliminer la possibilité de co-contractions sous-tendant ces adaptations, étant donné que le patron des EMG était approprié pour compenser chaque condition de champ de force. En parallèle, les neurones de M1 ont montré des changements systématiques dans leurs activités, sur le plan unitaire et populationnel, dans chaque condition de champ de force, signalant les changements requis dans la direction, l’amplitude et le décours temporel de la sortie de force musculaire nécessaire pour compenser les 5 conditions de champs de force. Les changements dans le patron de réponse pour chaque champ de force étaient assez cohérents entre les divers neurones de M1, ce qui suggère que la plupart des neurones de M1 contribuent à la compensation de toutes les conditions de champs de force, conformément aux prédictions du modèle MOSAIC. Aussi, cette modulation de l’activité neuronale ne supporte pas l’hypothèse d’une organisation fortement modulaire de M1. / In this thesis, we addressed motor control by two experimental approaches: psychophysical studies in human subjects and neurophysiological recordings in non-human primates. We identified unresolved issues concerning interference in motor learning during adaptation of subjects to two or more anti-correlated force fields. We designed paradigms in which arbitrary color stimuli provided contextual cues that allowed subjects to predict the nature of impending external force fields before encountering them physically during arm movements. This contextual knowledge helped to facilitate adaptation to the force fields by reducing this interference. According to one computational model of motor learning (MOdular Selection And Identification model for Control; MOSAIC), the color context cues made it easier for subjects to build “internal models” of each force field, to recall them and to switch between them with minimal interference. In our first experiment, four groups of human subjects performed elbow flexion/extension movements against two anti-correlated viscous force fields. We combined two different colors for the computer monitor background with two forces: resistive (Vr) and assistive (Va). The first two groups were control subjects. In those subjects, the color of the computer monitor changed at regular intervals but the force field remained constant; Vr was presented to the first group while the second group only experienced Va. As a result, the color cues were irrelevant in the two control groups. All control subjects adapted well to the single experienced force field (Vr or Va). In the two experimental groups, in contrast, the anti-correlated force fields and the monitor colors changed repeatedly between short blocks of trials. In the first experimental group (Reliable-cue subjects), there was a consistent relationship between the force and the stimulus (color of the monitor) - the red colour always signalled the resistive force while the green colour always signalled the assistive force. Adaptation to the two anti-correlated forces for the Reliable-cue group was significant during 10 days of training and almost as good as in the Irrelevant-cue groups who only experienced one of the two force fields. Furthermore, the Reliable-cue subjects quickly demonstrated predictive adaptive changes in their motor output whenever the monitor color changed, even during their first day of training, showing that they could use the reliable color context cues to recall the appropriate motor skills. In contrast, the monitor color also changed regularly between red and green in the second experimental group, but the force fields were not consistently associated with the color cue (Unreliable-cue group). These subjects took longer to adapt to the two force fields than the other three groups, and could not use the unreliable color cue change to make predictive changes to their motor output. Nevertheless, all Unreliable-cue subjects developed an ingenious strategy of making a specific “default” arm movement to probe the type of force field they would encounter in the first trial after the monitor color changed and used the proprioceptive feedback about the nature of the field to make appropriate predictive changes to their motor output for the next few trials, until the monitor color changed again, signifying the possibility of a change in force fields. This strategy was effective since the force remained constant in each short block of trials while the monitor color remained unchanged. This showed that the Unreliable-cue subjects were able to extract implicit and explicit information about the structure of the task from the color stimuli and use that knowledge to reduce interference when adapting to anti-correlated forces. The results of this first study encouraged us to advance our understanding of how subjects can recall multiple motor skills coupled to color context stimuli can be recalled, and how this phenomenon can be reflected by the neuronal activity in monkeys. Our aim was to elucidate how neurons of primary motor cortex (M1) can contribute to adaptive compensation for a wide range of different external forces during single-joint elbow flexion/extension movements. At the same time, we aimed to test the hypothesis evoked in the MOSAIC model, whereby multiple controller modules located in the cerebellum may predict each context and produce appropriate adaptive output signals for a small range of task conditions. Also, according to this hypothesis, M1 neurons may receive inputs from many specialized cerebellar controllers and show appropriate response modulations for a wide range of task conditions. We trained two monkeys to adapt their flexion/extension elbow movements against 5 different force-field conditions: null field without any external force disturbance, two anti-correlated viscous forces (assistive and resistive), which depended on movement speed and resembled that used in the human psychophysical study, a resistive elastic force which depended on elbow-joint position and finally, a visco-elastic field that was the linear sum of the elastic and viscous forces field. Each force field was reliably coupled to 5 different computer monitor background colors. The monkeys properly adapted to the 5 different force-field conditions and used the color context cues to recall the corresponding motor skill for the force field associated with each color, so that they could make predictive changes to their motor output before they physically encountered the force fields. EMG recordings eliminated the possibility that a co-contraction strategy was used by the monkeys to adapt to the force fields, since the EMG patterns were appropriate to compensate for each force-field condition. In parallel, M1 neurons showed systematic changes in their activity at the single-neuron and population level in each force-field condition that could signal the required changes in the direction, magnitude and time course of muscle force output required to compensate for the 5 force-field conditions. The patterns of response changes in each force field were consistent enough across M1 neurons to suggest that most M1 neurons contributed to the compensation for all force field conditions, in line with the predictions of the MOSAIC model. Also, these response changes do not support a strongly modular organization for M1.

The Theses deals with applications of mathematical knowledge in teaching biology and it is divided into four chapters. Each chapter is dedicated to another application: 1. Application of conditional probability in medical diagnostics, 2. Application of exponential function in population ecology, 3. Application of logic functions in mathematical modelation of neuron and 4. Aplication of binomial theorem and binomial distribution in genetics. Each application contains solved problems, a worksheet for students and a solution for each worksheet. Two application (1. and 2.) have been tested in teaching and as an assessment of my lessons students filled questionnaires. Results of these questionnaires are processed in the end of these chapters. This Thesis can be used in teaching or self-studying.

Spinal neurons are important in several aspects motor control. For example, the neurons essential for locomotor movements reside in the ventral spinal cord. In this thesis, different motor control functions are being related to neuronal populations defined by their common expression of a gene. First, a targeted disruption of the gene for vesicular glutamate transporter 2 (Vglut2/ Slc17a6) is described. The mutant animals die at birth because of their inability to breathe. The neuronal network in the brainstem, responsible for inspiration, was shown to become non-functional by the targeted deletion of Vglut2. To our surprise, it was still possible to induce rhythmic activity with normal left/right alternation in spinal cords isolated from VGLUT2-null embryos. Inconsistent reports of Vglut1 expression in the spinal cord made us re-evaluate the Vglut1 and Vglut2 expressions. While Vglut2 expression was widespread in the spinal cord, Vglut1 expression was restricted to a few cells dorsal to the central canal.  Taken together, the data suggest that, glutamatergic signaling is mandatory to drive the bilateral breathing, but not needed for coordination of basal alternating spinal locomotor rhythm. Next, a screen for genes with restricted ventral expression was made. Some of the genes found could be connected to the characteristics of specific neuronal cell populations. For example, fast motor neurons were shown to express the genes Calca and Chodl. Further, we found the Chrna2 expression selectively in putative Renshaw cells. It seems likely that the gene product, the alpha2 subunit of the nicotinergic receptor, could be linked to the unique connection of motor neurons to Renshaw cells. We used the Chrna2 promoter to drive expression of Cre recombinase in a transgenic mouse. The Cre activity was present in most neurons labeled with Renshaw cell markers, which should make it a useful tool for functional studies of this population. The studies presented here show how the genes expressed in subsets of neurons can be used to target populations of neurons for functional studies of neuronal systems.

Denna uppsatsen undersöker varför våld i film är så utbrett och populärt. Jag använder mig av ett evolutionärbiologiskt perspektiv, som tidigare inte använts vid filmforskning i så stor utsträckning. Istället har filmforskning och andra studier på människans beteende oftast utgått ifrån ett socialkonstruktivistiskt perspektiv. Uppsatsen är av typen ”forskningsöversikt” vilket innebär att jag inte har gjort någon klassisk analys på ett filmiskt verk eller empiriskt material, utan har sammanställt en stor mängd forskning från andra forskare. För att kunna svara på min frågeställning har jag behövt forska inom mänsklig evolution, audiovisuell perception samt våld i film. Den viktigaste bokenför uppsatsen kom att bli Torben Grodals Embodied Visions. Huvudfrågan i min frågeställning löd: Vilka faktorer ligger enligt tidigare forskning bakom populariteten av våldsskildringar?Svar: 1. Film är utformad för att påverka oss, aktivera känslor. 2. Det som påverkar ossallra mest är de bilder som påminner om de djupast rotade mekanismerna inuti oss. 3. Av evolutionära skäl ingår våld och aggressivitet bland dessa djupt rotade mekanismer. / This paper examines why violence in films is so widespread and popular. I use an evolutionary biological perspective, that has not previously been used in film research as much. Instead, the film research and other studies on human behavior usually originate from a social constructionist perspective. The essay is a "research review", which means that I have not made a classic analysis of a cinematic work or empirical data, but have collected a large amount of research from other scientists. To be able to answer my question, I had to do research in human evolution, audio visual perception and violence in film. The most important book for the essay came to be Torben Grodals Embodied Visions. The main question in my research question was: What factors are in accordance with previous research behind the popularity of graphic violence? Answer: 1. Film is designed to affect us, activating emotions. 2. what affects us the most is images that reminds us of deeply rooted mechanisms inside us. 3. of evolutionary reasons, violence and aggression are among these deep-rooted mechanisms.