Physical Activity and Fitness in Children with Developmental Disorder

Introduction: Developmental coordination disorder (DCD) is a prevalent condition characterized by poor motor proficiency that interferes with a child‟s activities of daily living. Children with DCD often experience compromised health-related fitness components such as cardiorespiratory fitness (CRF). Purpose: To better understand the physical activity and fitness characteristics of children with probable DCD (pDCD), with a particular focus on CRF. Specifically: (1) to present a synopsis of current literature; (2) to determine the longitudinal trajectories of CRF; (3) to compare the submaximal CRF of children with and without pDCD. Methods: A comprehensive, systematic literature review was conducted of the recent available data on fitness and physical activity and pDCD (Chapter 2). This review provided the background for the other two studies included in this thesis. In Chapter 3, a prospective cohort design was used to assess how CRF in children with pDCD changes over time (56 months) relative to a group of typically developing controls. Using a nested-case control design, 63 subjects with pDCD and 63 matched controls from the larger sample were recruited to participate in the lab-based component of the study (Chapter 4). In this investigation CRF was examined using the oxygen cost of work (VO2) during an incremental test on a cycle ergometer. Results: The literature review showed that fitness parameters, including CRF and physical activity levels, were consistently reduced in children with pDCD. Chapter 3 demonstrated that the difference in CRF between children with pDCD and typically developing children is substantial, and that it tends to increase over time. Results from VO2 assessments showed that children with pDCD utilized more oxygen to sustain the same submaximal workloads compared to typically developing children. Conclusions: Findings from this thesis have made several important contributions to our understanding of children with pDCD. Since differences in CRF between children with and without pDCD tend to worsen over time, this adds to the argument that interventions intended to improve CRF may be appropriate for children with motor difficulties. This thesis also presented the first evidence suggesting that DCD involves higher energy expenditure, and could help explain why children with pDCD perform poorly on tasks requiring CRF.

Reliability of muscle fiber conduction velocity in the tibialis anterior

This document could not have been completed without the hard work of a number of individuals. First and foremost, my supervisor, Dr. David Gabriel deserves the utmost recognition for the immense effort and time spent guiding the production of this document through the various stages of completion. Also, aiding in the data collection, technical support, and general thought processing were Lab Technician Greig Inglis and fellow members of the Electromyographic Kinesiology Laboratory Jon Howard, Sean Lenhardt, Lara Robbins, and Corrine Davies-Schinkel. The input of Drs. Ted Clancy, Phil Sullivan and external examiner Dr. Anita Christie, all members ofthe assessment committee, was incredibly important and vital to the completion of this work. Their expertise provided a strong source of knowledge and went to ensure that this project was completed at exemplary level. There were a number of other individuals who were an immense help in getting this project off the ground and completed. The donation of their time and efforts was very generous and much needed in order to fulfill the requirements needed for completion of this study. Finally, I cannot exclude the contributions of my family throughout this project especially that of my parents whose support never wavers.

Elucidation of the Retinoid Signalling Pathway Involved in Axon Guidance in Lymnaea stagnalis and Xenopus laevis

The vitamin A metabolite, retinoic acid (RA), is known to play a crucial role in several developmental processes including axial patterning and differentiation. More recently, RA has been implicated in the regenerative process acting through its classical signaling pathway, the nuclear receptors, retinoic acid receptor (RAR) and retinoid X receptor (RXR), to mediate gene transcription. Moreover, RA has been shown to act as a guidance molecule for growth cones of regenerating motorneurons of the pond snail, Lymnaea stagnalis. Our lab has recently shown that RA can induce this morphological response independent of nuclear transcription, however, the role of the retinoid receptors in RA-induced chemoattraction is still unknown. Here, I show that the retinoid receptors, RXR and RAR, may mediate the growth cones response to the metabolically active retinoic acid isomers, all-trans and 9-cis RA, in Lymnaea stagnalis. Data presented here show that both an RXR and RAR antagonist can block growth cone turning in response to application of both isomers. Because no prior investigations have shown growth cone turning of individual vertebrate neurons, I aimed to show that both retinoic acid isomers were capable of inducing growth cone turning of embryonic spinal cord neurons in the frog, Xenopus laevis. For the first time in Xenopus, I showed that both all-trans and 9-cis RA were able to induce significantly more neurite outgrowth from cultured embryonic spinal cord neurons and induce positive growth cone turning of individual growth cones. In addition, I showed that the presence of the RXR antagonist, HX531, blocked 9-cis RA-induced growth cone turning and the RARβ antagonist, LE135, blocked all-trans RA-induced growth cone turning in this species. Evidence provided here shows for the first time, conservation of retinoic acid-induced growth cone turning in a vertebrate model system. In addition, these data show that the receptors involved in this morphological response may be the same in vertebrates and invertebrates.

An Investigation of the Role of Attention in the Cross-Race Effect: An Ecological Approach

The current set of studies was conducted to examine the cross-race effect (CRE), a phenomenon commonly found in the face perception literature. The CRE is evident when participants display better own-race face recognition accuracy than other-race recognition accuracy (e.g. Ackerman et al., 2006). Typically the cross-race effect is attributed to perceptual expertise, (i.e., other-race faces are processed less holistically; Michel, Rossion, Han, Chung & Caldara, 2006), and the social cognitive model (i.e., other-race faces are processed at the categorical level by virtue of being an out-group member; Hugenberg, Young, Bernstein, & Sacco, 2010). These effects may be mediated by differential attention. I investigated whether other-race faces are disregarded and, consequently, not remembered as accurately as own-race (in-group) faces. In Experiment 1, I examined how the magnitude of the CRE differed when participants learned individual faces sequentially versus when they learned multiple faces simultaneously in arrays comprising faces and objects. I also examined how the CRE differed when participants recognized individual faces presented sequentially versus in arrays of eight faces. Participants’ recognition accuracy was better for own-race faces than other-race faces regardless of familiarization method. However, the difference between own- and other-race accuracy was larger when faces were familiarized sequentially in comparison to familiarization with arrays. Participants’ response patterns during testing differed depending on the combination of familiarization and testing method. Participants had more false alarms for other-race faces than own-race faces if they learned faces sequentially (regardless of testing strategy); if participants learned faces in arrays, they had more false alarms for other-race faces than own-races faces if ii i they were tested with sequentially presented faces. These results are consistent with the perceptual expertise model in that participants were better able to use the full two seconds in the sequential task for own-race faces, but not for other-race faces. The purpose of Experiment 2 was to examine participants’ attentional allocation in complex scenes. Participants were shown scenes comprising people in real places, but the head stimuli used in Experiment 1 were superimposed onto the bodies in each scene. Using a Tobii eyetracker, participants’ looking time for both own- and other-race faces was evaluated to determine whether participants looked longer at own-race faces and whether individual differences in looking time correlated with individual differences in recognition accuracy. The results of this experiment demonstrated that although own-race faces were preferentially attended to in comparison to other-race faces, individual differences in looking time biases towards own-race faces did not correlate with individual differences in own-race recognition advantages. These results are also consistent with perceptual expertise, as it seems that the role of attentional biases towards own-race faces is independent of the cognitive processing that occurs for own-race faces. All together, these results have implications for face perception tasks that are performed in the lab, how accurate people may be when remembering faces in the real world, and the accuracy and patterns of errors in eyewitness testimony.

Hyperosmotic Stress and the Impact on Metabolite Formation and Redox Balance in Saccharomyces cerevisiae and Saccharomyces bayanus strains

Wine produced using an appassimento-type process represents a new and exciting innovation for the Ontario wine industry. This process involves drying grapes that have already been picked from the vine, which increases the sugar content due to dehydration and induces a variety of changes both within and on the surface of the grapes. Increasing sugar contents in musts subject wine yeast to conditions of high osmolarity during alcoholic fermentations. Under these conditions, yeast growth can be inhibited, target alcohol levels may not be attained and metabolic by-products of the hyperosmotic stress response, including glycerol and acetic acid, may impact wine composition. The further metabolism of acetic acid to acetylCoA by yeast facilitates the synthesis of ethyl acetate, a volatile compound that can also impact wine quality if present in sufficiently high concentrations. The first objective of this project was to understand the effect of yeast strain and sugar concentration on fermentation kinetics and metabolite formation, notably acetic acid and ethyl acetate, during fermentation in appassimento-type must. Our working hypotheses were that (1) the natural isolate Saccharomyces bayanus would produce less acetic acid and ethyl acetate compared to Saccharomyces cerevisiae strain EC-1118 fermenting the high and low sugar juices; (2) the wine produced using the appassimento process would contain higher levels of acetic acid and lower levels of ethyl acetate compared to table wine; (3) and the strains would be similar in the kinetic behavior of their fermentation performances in the high sugar must. This study determined that the S. bayanus strain produced significantly less acetic acid and ethyl acetate in the appassimento wine and table wine fermentations. Differences in acetic acid and ethyl acetate production were also observed within strains fermenting the two sugar conditions. Acetic acid production was higher in table wine fermented by S. bayanus as no acetic acid was produced in appassimento-style wine, and 1.4-times higher in appassimento wine fermented by EC-1118 over that found in table wine. Ethyl acetate production was 27.6-times higher in table wine fermented by S. bayanus, and 5.2-times higher by EC-1118, compared to that in appassimento wine. Sugar utilization and ethanol production were comparable between strains as no significant differences were determined. The second objective of this project was to bring a method in-house for measuring the concentration of pyridine nucleotides, NAD+, NADP+, NADH and NADPH, in yeast cytosolic extract. Development of this method is of applicative interest for our lab group as it will enable the redox balance of the NAD+/ NADH and NADP+/ NADPH systems to be assessed during high sugar fermentations to determine their respective roles as metabolic triggers for acetic acid production. Two methods were evaluated in this study including a UV-endpoint method using a set of enzymatic assay protocols outlined in Bergmeyer (1974) and a colorimetric enzyme cycling method developed by Sigma-Aldrich® using commercial kits. The former was determined to be limited by its low sensitivity following application to yeast extract and subsequent coenzyme analyses, while the latter was shown to exhibit greater sensitivity. The results obtained from the kits indicated high linearity, accuracy and precision of the analytical method for measuring NADH and NADPH, and that it was sensitive enough to measure the low coenzyme concentrations present in yeast extract samples. NADtotal and NADPtotal concentrations were determined to be above the lower limit of quantification and within the range of the respective calibration curves, making this method suitable for our research purposes.

Process Improvement in Phlebotomy

This study has two main objectives. First, the phlebotomy process at the St. Catharines Site of the Niagara Health System is investigated, which starts when an order for a blood test is placed, and ends when the specimen arrives at the lab. The performance measurement is the flow time of the process, which reflects concerns and interests of both the hospital and the patients. Three popular operational methodologies are applied to reduce the flow time and improve the process: DMAIC from Six Sigma, lean principles and simulation modeling. Potential suggestions are provided for the St. Catharines Site, which could result in an average of seven minutes reduction in the flow time. The second objective addresses the fact that these three methodologies have not been combined before in a process improvement effort. A structured framework combining them is developed to benefit future study of phlebotomy and other hospital processes.

Nonsuicidal Self-Injury and Suicidal Risk: An Examination among Young Adults

Nonsuicidal self-injury (NSSI), which refers to the direct and deliberate destruction of bodily tissue in the absence of suicidal intent, is a serious and widespread mental health concern. Although NSSI has been differentiated from suicidal behavior on the basis of non-lethal intent, research has shown that these two behaviors commonly co-occur. Despite increased research on the link between NSSI and suicidal behavior, however, little attention has been given as to why these two behaviors are associated. My doctoral dissertation specifically addressed this gap in the literature by examining the link between NSSI and several measures of suicidal risk (e.g., suicidal ideation, suicidal attempts, pain tolerance) among a large sample of young adults. The primary goal of my doctoral research was to identify individuals who engaged in NSSI at risk for suicidal ideation and attempts, in an effort to elucidate the processes through which psychosocial risk, NSSI, and suicidal risk may be associated. Participants were drawn from a larger sample of 1153 undergraduate students (70.3% female) at a mid-sized Canadian University. In study one, I examined whether increases in psychosocial risk and suicidal ideation were associated with changes in NSSI engagement over a one year period. Analyses revealed that beginners, relapsed injurers, and persistent injurers were differentiated from recovered injurers and desisters by increases in psychsocial risk and suicidal ideation over time. In study two, I examined whether several NSSI characteristics (e.g., frequency, number of methods) were associated with suicidal risk using latent class analysis. Three subgroups of individuals were identified: 1) an infrequent NSSI/not high risk for suicidal behavior group, 2) a frequent NSSI/not high risk for suicidal behavior group, and 3) a frequent NSSI/high risk for suicidal behavior group. Follow-up analyses indicated that individuals in the frequent NSSI/high risk for suicidal behavior group met the clinical cutoff score for high suicidal risk and reported significantly greater levels of suicidal ideation, attempts, and risk for future suicidal behavior as compared to the other two classes. Class 3 was also differentiated by higher levels of psychosocial risk (e.g., depressive symptoms, social anxiety) relative to the other two classes, as well as a comparison group of non-injuring young adults. Finally, in study three, I examined whether NSSI was associated with pain tolerance in a lab-based task, as tolerance to pain has been shown to be a strong predictor of suicidal risk. Individuals who engaged in NSSI to regulate the need to self-punish, tolerated pain longer than individuals who engaged in NSSI but not to self-punish and a non-injuring comparison group. My findings offer new insight into the associations among psychosocial risk, NSSI, and suicidal risk, and can serve to inform intervention efforts aimed at individuals at high risk for suicidal behavior. More specifically, my findings provide clinicians with several NSSI-specific risk factors (e.g., frequent self-injury, self-injuring alone, self-injuring to self-punish) that may serve as important markers of suicidal risk among individuals engaging in NSSI.

Working Together: Librarian and Student Collaboration Through Active Learning in a Library Eclassroom

Active learning strategies based on several learning theories were incorporated during instruction sessions for second year Biological Sciences students. The instructional strategies described in this paper are based primarily on sociocultural and collaborative learning theory, with the goal being to expand the relatively small body of literature currently available that discusses the application of these learning theories to library instruction. The learning strategies employed successfully involved students in the learning process ensuring that the experiences were appropriate and effective. The researchers found that, as a result of these strategies (e.g. teaching moments based on the emerging needs of students) students’ interest in learning information literacy was increased and students interacted with information given to them as well as with their peers. Collaboration between the Librarians, Co-op Student and Senior Lab Instructor helped to enhance the learning experience for students and also revealed new aspects of the active learning experiences. The primary learning objective, which was to increase the students’ information skills in the Biological Sciences, was realized. The advantages of active learning were realized by both instructors and students. Advantages for students attained during these sessions include having their diverse learning styles addressed; increased interaction with and retention of information; increased responsibility for their own learning; the opportunity to value not only the instructors, but also themselves and their peers as sources of authority and knowledge; improved problem solving abilities; increased interest and opportunities for critical thinking, as a result of the actively exchanging information in a group. The primary advantage enjoyed by the instructors was the opportunity to collaborate with colleagues to reduce the preparation required to create effective library instruction sessions. Opportunities for further research were also discovered, including the degree to which “social loafing” plays a role in collaborative, active learning.

Evolution of Various Library Instruction Strategies: Using Student Feedback to Create and Enhance Online Active Learning Assignments

This case study traces the evolution of library assignments for biological science students from paper-based workbooks in a blended (hands-on) workshop to blended learning workshops using online assignments to online active learning modules which are stand-alone without any face-to-face instruction. As the assignments evolved to adapt to online learning supporting materials in the form of PDFs (portable document format), screen captures and screencasting were embedded into the questions as teaching moments to replace face-to-face instruction. Many aspects of the evolution of the assignment were based on student feedback from evaluations, input from senior lab demonstrators and teaching assistants, and statistical analysis of the students’ performance on the assignment. Advantages and disadvantages of paper-based and online assignments are discussed. An important factor for successful online learning may be the ability to get assistance.

Standardization of cytokine flow cytometry assays

Affiliation: Faculté de médecine, Université de Montréal & CANVAC

The Origin and Stimuli Implicated in the Expression of Nestin(+) Cardiac Myocyte-like Cells in the Ischemic Heart

Nos études ont démontrées que la formation de la cicatrice et la guérison sont associées avec l’apparition de cellules de type myocytes cardiaques nestine(+) dans la région péri-infarcie. Présentement, l’étude examine le mécanisme, tel que l’hypoxie ou les hormones neuronales, possiblement impliqué dans leur recrutement et de dévoiler leur origine cellulaire. La présence de ces cellules a été détectée dans les coeurs infarcies d’une semaine et maintenue après neuf mois suite à une sujétion coronaire complète. Aussi, ces cellules de type myocytes cardiaques nestine(+) ont été observées dans le coeur infarci humain. L’hypoxie représente un événement prédominant suite à un infarctus de myocarde, mais l’exposition des rats normaux à un environnement hypoxique n’a pas pu promouvoir l’apparition de ces cellules. Autrement, l’infusion de l’agoniste -adrénergique non-sélectif isoprotérénol (ISO) dans les rats adultes Sprague-Dawley a augmenté la protéine nestine dans le ventricule gauche et a été associé avec la réapparition de cellules de type myocytes cardiaques nestine(+). Cela représente possiblement un effet secondaire suite à la nécrose des myocytes cardiaques par l’administration d’isoprotérénol. Dernièrement, on a identifié une sous-population de cellules nestine(+) dans le coeur normal du rat qui co-exprime les marqueurs de cellules cardiaques progénitrices Nkx-2.5 et GATA-4. Cette sous-population de cellules nestine/Nkx-2.5/GATA-4 pourrait représenter des substrats cellulaires qui puissent se différentier en cellules de type myocytes cardiaques nestine(+) suite à une ischémie. Mots clés: nestine, isoprotérénol, nécrose, cellule souche, cellule progénitrice, myocyte cardiaque

Systematic analysis of protein complexes involved in the human RNA polymerase II machinery

La transcription, la maturation d’ARN, et le remodelage de la chromatine sont tous des processus centraux dans l'interprétation de l'information contenue dans l’ADN. Bien que beaucoup de complexes de protéines formant la machinerie cellulaire de transcription aient été étudiés, plusieurs restent encore à identifier et caractériser. En utilisant une approche protéomique, notre laboratoire a purifié plusieurs composantes de la machinerie de transcription de l’ARNPII humaine par double chromatographie d’affinité "TAP". Cette procédure permet l'isolement de complexes protéiques comme ils existent vraisemblablement in vivo dans les cellules mammifères, et l'identification de partenaires d'interactions par spectrométrie de masse. Les interactions protéiques qui sont validées bioinformatiquement, sont choisies et utilisées pour cartographier un réseau connectant plusieurs composantes de la machinerie transcriptionnelle. En appliquant cette procédure, notre laboratoire a identifié, pour la première fois, un groupe de protéines, qui interagit physiquement et fonctionnellement avec l’ARNPII humaine. Les propriétés de ces protéines suggèrent un rôle dans l'assemblage de complexes à plusieurs sous-unités, comme les protéines d'échafaudage et chaperonnes. L'objectif de mon projet était de continuer la caractérisation du réseau de complexes protéiques impliquant les facteurs de transcription. Huit nouveaux partenaires de l’ARNPII (PIH1D1, GPN3, WDR92, PFDN2, KIAA0406, PDRG1, CCT4 et CCT5) ont été purifiés par la méthode TAP, et la spectrométrie de masse a permis d’identifier de nouvelles interactions. Au cours des années, l’analyse par notre laboratoire des mécanismes de la transcription a contribué à apporter de nouvelles connaissances et à mieux comprendre son fonctionnement. Cette connaissance est essentielle au développement de médicaments qui cibleront les mécanismes de la transcription.

The role of NHL-2 in regulating C.elegans P granule function

La divison cellulaire asymétrique est un processus essentiel qui permet aux cellules souches de s’auto-renouveller et de produire une cellule fille destinée à la différenciation. La lignée germinale de C. elegans, totipotente et immortelle, est une lignée de cellules souches qui contient des organites ribonucléoprotéiques appelés granules P. Au cours du développement ces derniers sont toujours localisés spécifiquement dans les cellules précurseurs de la lignée germinals, suggérant qu’ils sont des déterminants de la lignée germinale. De façon intéressante, des granules ribonucléoprotéiques, comme les P bodies impliqués dans le contrôle post-transcriptionnel, ont été observés chez tous les organismes. Néanmoins, la fonction précise des granules P de C. elegans est inconnue. Récemment, notre laboratoire a montré que NHL-2, un homologue de Mei-P26 de Drosophile, colocalise avec les granules P dans des embryons précoces et joue un rôle dans la division cellulaire asymétrique et dans la polarité cellulaire. Tous les granules P contiennent NHL- 2, ce qui nous a mené à poser l’hypothèse que NHL-2 régule la biogenèse et la fonction des granules P. Nous avons testé cette hypothèse par imagerie et quantification de l'intensité de PGL-1, un composant essentiel des granules P, dans des embryons fixés. Nos résultats montrent que dans des embryons mutants pour nhl-2 il y a une réduction du nombre de granules P, de l'intensité de fluorescence moyenne (IFM) et de l'intensité de fluorescence total (IFT) de PGL-1. Une analyse plus poussée a montré qu'il existe deux populations distinctes d’embryons mutants pour nhl-2 : l’une présente une intensité de PGL-1 comparable à celle d’une population sauvage alors que le second groupe présente une forte réduction des quantités de PGL-1 et est comparable à des mutants pour pgl-1. Cette variabilité est aussi observée dans le phénotype de stérilité de nhl-2 mutant à des températures élevées. Globalement, nos résultats suggèrent que la perte de fonction de NHL-2 perturbe la prolifération des cellules germinales ainsi que la formation et/ou la stabilité des granules P au cours des étapes précoces du développement des précurseurs de la lignée germinals. D’autre part, ils suggèrent que la fonction de NHL-2 pourrait être partiellement redondants avec les autres régulateurs de la stabilité des granules P. Mots-clés : Granules P, NHL-2, Cellules germinals.

Mécanismes moléculaires impliqués dans la régulation de l’acide polysialique (PSA) dans le néocortex visuel des souris durant la maturation des synapses GABAergiques

Le fonctionnement du cortex cérébral nécessite l’action coordonnée de deux des sous-types majeurs de neurones, soient les neurones à projections glutamatergiques et les interneurones GABAergiques. Les interneurones GABAergiques ne constituent que 20 à 30% des cellules corticales par rapport au grand nombre de neurones glutamatergiques. Leur rôle est toutefois prépondérant puisqu’ils modulent fortement la dynamique et la plasticité des réseaux néocorticaux. Il n’est donc pas surprenant que les altérations de développement des circuits GABAergiques soient associées à plusieurs maladies du cerveau, incluant l’épilepsie, le syndrome de Rett et la schizophrénie. La compréhension des mécanismes moléculaires régissant le développement des circuits GABAergiques est une étape essentielle menant vers une meilleure compréhension de la façon dont les anormalités se produisent. Conséquemment, nous nous intéressons au rôle de l’acide polysialique (PSA) dans le développement des synapses GABAergiques. PSA est un homopolymère de chaînons polysialylés en α-2,8, et est exclusivement lié à la molécule d’adhésion aux cellules neuronales (NCAM) dans les cerveaux de mammifères. PSA est impliqué dans plusieurs processus développementaux, y compris la formation et la plasticité des synapses glutamatergiques, mais son rôle dans les réseaux GABAergiques reste à préciser. Les données générées dans le laboratoire du Dr. Di Cristo démontrent que PSA est fortement exprimé post- natalement dans le néocortex des rongeurs, que son abondance diminue au cours du développement, et, faits importants, que son expression dépend de l’activité visuelle i et est inversement corrélée à la maturation des synapses GABAergiques. La présente propose de caractériser les mécanismes moléculaires régulant l’expression de PSA dans le néocortex visuel de la souris. Les enzymes polysialyltransférases ST8SiaII (STX) et ST8SiaIV (PST) sont responsables de la formation de la chaîne de PSA sur NCAM. En contrôlant ainsi la quantité de PSA sur NCAM, ils influenceraient le développement des synapses GABAergiques. Mon projet consiste à déterminer comment l’expression des polysialyltransférases est régulée dans le néocortex visuel des souris durant la période post-natale; ces données sont à la fois inconnues, et cruciales. Nous utilisons un système de cultures organotypiques dont la maturation des synapses GABAergiques est comparable au modèle in vivo. L’analyse de l’expression génique par qPCR a démontré que l’expression des polysialyltransférases diminue au cours du développement; une baisse majeure corrélant avec l’ouverture des yeux chez la souris. Nous avons de plus illustré pour la première fois que l’expression de STX, et non celle de PST, est activité-dépendante, et que ce processus requiert l’activation du récepteur NMDA, une augmentation du niveau de calcium intracellulaire et la protéine kinase C (PKC). Ces données démontrent que STX est l’enzyme régulant préférentiellement le niveau de PSA sur NCAM au cours de la période post-natale dans le cortex visuel des souris. Des données préliminaires d’un second volet de notre investigation suggèrent que l’acétylation des histones et la méthylation de l’ADN pourraient également contribuer à la régulation de la transcription de cette enzyme durant le développement. Plus d’investigations seront toutefois nécessaires afin de confirmer cette hypothèse. En somme, la connaissance des mécanismes par lesquels l’expression des ii polysialyltransférases est modulée est essentielle à la compréhension du processus de maturation des synapses GABAergiques. Ceci permettrait de moduler pharmacologiquement l’expression de ces enzymes; la sur-expression de STX et/ou PST pourrait produire une plus grande quantité de PSA, déstabiliser les synapses GABAergiques, et conséquemment, ré-induire la plasticité cérébrale.

Régulation transcriptionelle du développement de l'hypothalamus chez l'amphibien

Le noyau paraventriculaire (PVN) de l'hypothalamus régule une série de phénomènes physiologiques incluant l'équilibre énergétique et la pression artérielle. Nous avons identifié une cascade de facteurs de transcription qui contrôle le développement du PVN. SIM1 et OTP agissent en parallèle pour contrôler la différenciation d'au moins cinq types de neurones identifiables par la production d'OT, AVP, CRH, SS et TRH. Ces Facteurs de transcriptions contrôlent le développement des lignées CRH, AVP et OT en maintenant l'expression de Brn2 qui à son tour est nécessaire pour la différenciation terminale de ces neurones. L'analyse du transcriptome du PVN nous a permis d'identifier plusieurs gènes qui ont le potentiel de contrôler le développement du PVN. Nous voulons développer un paradigme de perte de fonction qui permettrait l'étude de ces gènes candidats sur une grande échelle. Le but de ce projet est de caractériser le PVN en développement de l'amphibien en vue de l'utilisation de ce modèle pour des études fonctionnelles. Nous avons cloné des fragments de cDNA de Sim1, OTP, Brn2, Sim2, CRH, Ot, AVP et TRH à partir de l'ARN total de Xenopus Laevis. Nous avons adapté notre technique d'hybridation in situ pour caractériser l'expression de ces gènes chez l'amphibien aux stades 33-39, 44, 51, 54, 60, et chez l'adulte. Résultats. Les Facteurs de transcription Sim1, OTP, et Brn2 commencent à être exprimés dans le PVN prospectif au stade 33. L'expression des marqueurs de différenciation terminale devient détectable entre les stades 37 et 39. De façon intéressante, le PVN occupe initialement un domaine de forme globulaire puis à partir du stade 44 s'allonge le long de l’axe dorso-ventral. Cet allongement se traduit par une organisation en colonnes des cellules du PVN que nous n'avons pas observée chez les rongeurs. Le développement du PVN est conservé chez l'amphibien dans la mesure où la relation entre l'expression des facteurs de transcription et des marqueurs de différenciation terminale est conservée. Il existe par ailleurs des différences entre la topographie des PVN des mammifères et de l'amphibien. L'organisation en colonnes de cellules pourrait correspondre à des mouvements de migration tangentielle. Nous sommes maintenant en mesure de tester la fonction des facteurs de transcription dans le PVN par l'approche d'invalidation par morpholinos.