This thesis first explored the effects of concurrent spatial attention and working memory task performance on over-ground gait in healthy young and older adults. It then compared over-ground gait parameters and working memory performance in mild Alzheimer’s Disease (AD) and normal controls (NC) and investigated costs of dual-tasking on working memory performance and cadence during treadmill walking at preferred walking speed in the two groups. Furthermore, it explored these differences in AD and NC groups in relation to their subcortical hyperintensities (SH) that were rated using standardized scales on MRI. Reaction times and accuracy on working memory performance measures were collected under single and dual task conditions. Over-ground gait parameters were measured on an automated walkway. Costs of dual-tasking on gait parameters and working memory performance were measured at a constant velocity on a treadmill. The hypotheses that working memory influences gait performance and that a higher SH burden negatively influences over-ground gait and costs of dual-task conditions, were supported in a series of experiments. Gait slowed down while performing working memory and spatial attention tasks in young and older adults. Patients with mild AD, compared to NC, had a slower gait velocity, shorter stride length and lower cadence on the walkway. When the two groups were subdivided into higher and lower SH groups based on their median SH score, the NC group with lower SH burden walked significantly faster with a higher cadence and a longer stride length than the other three groups. Lastly, a higher SH burden negatively influenced working memory performance in NC while in mild AD patients, it had negative influences on adaptive changes in gait while dual-tasking. These results suggest that, in dual-task condition, SH interfere with processing speed in NC and on gait in AD. These findings provide new insights in to tradeoffs during dual tasking in relation to cerebrovascular disease. This has ecological implications because of the prevalence of small vessel disease in aging and dementia, may impact on predicting falls in AD.

This thesis first explored the effects of concurrent spatial attention and working memory task performance on over-ground gait in healthy young and older adults. It then compared over-ground gait parameters and working memory performance in mild Alzheimer’s Disease (AD) and normal controls (NC) and investigated costs of dual-tasking on working memory performance and cadence during treadmill walking at preferred walking speed in the two groups. Furthermore, it explored these differences in AD and NC groups in relation to their subcortical hyperintensities (SH) that were rated using standardized scales on MRI. Reaction times and accuracy on working memory performance measures were collected under single and dual task conditions. Over-ground gait parameters were measured on an automated walkway. Costs of dual-tasking on gait parameters and working memory performance were measured at a constant velocity on a treadmill. The hypotheses that working memory influences gait performance and that a higher SH burden negatively influences over-ground gait and costs of dual-task conditions, were supported in a series of experiments. Gait slowed down while performing working memory and spatial attention tasks in young and older adults. Patients with mild AD, compared to NC, had a slower gait velocity, shorter stride length and lower cadence on the walkway. When the two groups were subdivided into higher and lower SH groups based on their median SH score, the NC group with lower SH burden walked significantly faster with a higher cadence and a longer stride length than the other three groups. Lastly, a higher SH burden negatively influenced working memory performance in NC while in mild AD patients, it had negative influences on adaptive changes in gait while dual-tasking. These results suggest that, in dual-task condition, SH interfere with processing speed in NC and on gait in AD. These findings provide new insights in to tradeoffs during dual tasking in relation to cerebrovascular disease. This has ecological implications because of the prevalence of small vessel disease in aging and dementia, may impact on predicting falls in AD.

Le gène Notch3 code pour un récepteur transmembranaire hétérodimérique exprimé principalement dans les cellules musculaires lisses des petites artères. Les travaux de ces dernières années ont montré que le récepteur Notch3 joue un rôle clé dans la physiologie et la pathologie des petites artères. Chez la souris, Notch3 est requis pour l’intégrité structurale et fonctionnelle des artères de résistance en contrôlant l’identité artérielle, la maturation postnatale des cellules musculaires lisses et le tonus myogénique des artères de résistance. Chez l’Homme, les maladies des petites artères cérébrales (MPAC) regroupent un ensemble hétérogène de maladies parmi lesquelles un petit pourcentage, probablement encore sous-estimée, est héréditaire. A ce jour, très peu de gènes responsables de formes familiales de MPAC ont été identifiés. CADASIL est la forme familiale la plus fréquente de MPAC causée par des mutations du gène NOTCH3. Il s’agit de mutations extrêmement stéréotypées siégeant dans les répétitions EGF qui constituent le domaine extracellulaire de Notch3. Les résultats du laboratoire suggèrent fortement que l’effet pathogène de ces mutations résulte de l’acquisition par le récepteur muté d’une nouvelle fonction. Les deux objectifs de ce travail ont été : 1°) Tester l’hypothèse qu’il existe des maladies des petites artères cérébrales causées directement par une modification de l’activité du récepteur Notch3 2°) Identifier les effecteurs du récepteur Notch3 dans le contexte du développement et de la maturation des artères cérébrales Nous avons identifié chez une patiente présentant une MPAC distincte de CADASIL une nouvelle mutation siégeant dans le domaine d’hétérodimérisation du récepteur Notch3. In vitro, la mutation L1515P induit une activation ligand indépendante du récepteur Notch3. L’analyse biochimique suggère que cette activation est causée par une déstabilisation du domaine d’hétérodimérisation de Notch3.Nous avons réalisé une analyse du transcriptome des artères caudales de souris Notch3-/- et Notch3+/+. Cette analyse a permis d’identifier un groupe de 17 gènes régulés par Notch3 dans l’artère caudale ou les artères cérébrales. L’invalidation du facteur de transcription CSL/RBPJK dans les cellules musculaires lisses, pendant la période postnatale immédiate, phénocopie les altérations structurales et moléculaires observées chez les souris Notch3-/-. L’administration chez la souris d’un inhibiteur pharmacologique de la voie Notch a permis d’identifier 6 gènes (Grip2, Nrip2, Kv1.5, Pgam2, Susd5, Xirp1), en plus de Notch3, dont l’expression dans les artères cérébrales est rapidement diminuée par ce traitement. Nous avons ensuite concentré nos efforts sur le gène Grip2 dont l’expression était la plus fortement diminuée dans les différents modèles d’inactivation de la voie Notch. Grip2 était jusqu’alors connu pour son interaction avec les récepteurs au glutamate dans les neurones. Nous avons montré que Grip2 était également exprimé dans les cellules musculaires lisses vasculaires et identifié une isoforme vasculaire régulée spécifiquement par Notch3/CSL/RBPJK. L’analyse des souris Grip2neo/neo, exprimant une protéine Grip2 tronquée dans sa partie N-terminale, a révélé une atteinte sélective du tonus myogénique des artères cérébrales.En conclusion, nous avons démontré l’existence d’une mutation activatrice de NOTCH3 associée à une MPAC chez l’Homme. Nos résultats indiquent que dans le contexte de la maturation des artères cérébrales, la fonction de Notch3 est médiée par le facteur de transcription CSL/RBPJK dans les cellules musculaires lisses durant la période postnatale immédiate. Nous avons identifié plusieurs nouveaux effecteurs potentiels de Notch3 et validé l’un d’entre eux, Grip2, pour son implication dans les réponses myogéniques des artères cérébrales. Nous proposons que des mutations dans les gènes codant pour ces effecteurs puissent rendre compte de certaines formes monogéniques de MPAC. / Notch3 encodes a transmembrane receptor primarily expressed in arterial smooth muscle cells. Human and mouse genetics studies demonstrated that Notch3 is a key player in physiology and diseases of small vessels. Studies in mice revealed that Notch3 is required to generate functional arteries in regulating arterial differentiation, maturation of vascular smooth muscle cells and myogenic tone. Cerebral Autosomal Dominant Arteriopathy with Subcortical infarcts and Leukoencephalopathy (CADASIL) is the most frequent hereditary small vessels disease in human adults caused by NOTCH3 mutations. Pathogenic mutations lead to an odd number of cysteine residues within the NOTCH3 extracellular domain. Data from the laboratory suggest a model that invokes novel pathogenic roles from the mutant NOTCH3 protein. The main goals of this work are: 1°) To determine if there is small vessels disease caused by modification of Notch3 activity 2°) To identify Notch3 effectors involved in development and maturation of cerebral arteries We identified a novel heterozygous missense mutation (L1515P) in the heterodimerization domain of NOTCH3 in a patient with cerebral small vessel distinct from CADASIL. In vitro analysis showed that the L1515P mutant exhibits increased canonical NOTCH3 signaling in a ligand-independent manner. Biochemical analysis suggests that the mutation renders NOTCH3 hyperactive through destabilization of the heterodimer. Transcriptome analysis using tail arteries of Notch3-/- and Notch3+/+ mice identified a core set of 17 novel Notch3-regulated genes confirmed in tail or brain arteries. Postnatal deletion of RBP-Jκ in smooth muscle cells recapitulated the structural, functional, and molecular defects of brain arteries induced by Notch3 deficiency. Transient in vivo blockade of the Notch pathway with γ-secretase inhibitors uncovered, in addition to Notch3, 6 immediate responders, including the voltage-gated potassium channel Kv1.5, which opposes to myogenic constriction of brain arteries, and the glutamate receptor-interacting protein-2, with no previously established role in the cerebrovasculature. We identified a vascular smooth muscle cell isoform of Grip2. We showed that Notch3-RBP-Jκ specifically regulates this isoform. Finally, we found that cerebral arteries of glutamate receptor-interacting protein-2 mutant mice, which express an N-terminally truncated glutamate receptor-interacting protein-2, exhibited selective attenuation of pressure-induced contraction. In conclusion, we have demonstrated the existence of a NOTCH3 activating mutation associated with small vessels disease in human. Our results show that, in the context of cerebral arteries maturation, Notch3 functions are mediated by CSL/RBPJK transcription factor. We have identified several new Notch3 effectors and validated Grip2 as a novel regulator of myogenic tone in cerebral arteries. One can expect that mutations in these Notch3-regulated genes could be responsible of some monogenic form of small vessel diseases of the brain.

No description available.

[pt] O Comprometimento cognitivo vascular subcortical (CCVs) caracteriza-se, do ponto de vista cognitivo, por déficits em funções executivas em seus estágios iniciais, levando a dificuldades funcionais precoces na população acometida. Contudo, dados da literatura indicaram que a condição é frequentemente subdiagnosticada, o que pode ser atribuído, dentre outras razões, à falta de instrumentos de rastreio acurados para a condição, cujas características clínicas diferem dos quadros neurodegenerativos. Esta escassez foi evidenciada pela revisão sistemática realizada como parte do presente trabalho. O Teste Breve de Memória e Funções Executivas (BMET) foi desenvolvido por Brookes e cols. (2012) especificamente para a detecção destes quadros clínicos. O objetivo desta dissertação foi discorrer sobre o CCVs e adaptar o BMET para o uso na população acima dos 60 anos do Brasil. Para tanto, seguiram-se etapas de tradução, avaliação de juízes de linguística e de neuropsicologia, avaliação por representante do público-alvo, retrotradução e, por fim, aplicação à amostra. Foi conduzido um estudo piloto para verificar a aplicabilidade e fornecer dados psicométricos preliminares do teste. Conclui-se que o BMET é de simples e rápida aplicação, podendo ser administrado por diferentes profissionais da saúde e possivelmente adequado à atenção primária em saúde. Considerando o contexto atual de um rápido envelhecimento populacional e do consequente aumento na prevalência de agravos associados às faixas etárias avançadas, tais como os transtornos neurocognitivos, é de grande relevância a detecção precoce do CCVs visando ao planejamento de intervenções e medidas de profilaxia secundária, contribuindo assim, para a qualidade de vida da população idosa. / [en] Subcortical Vascular Cognitive Impairment (sVCI) is characterized, upon a cognitive perspective, by the presence of dysexecutive function in its initial stages, leading to early functional difficulties for sufferers of this condition. However, data has indicated that it is often underdiagnosed, which could be attributed to the lack of accurate screening tools for the disorder. This shortcoming has been evidenced by a systematic review conducted as part of the current project. The Brief Memory and Executive Test (BMET) has been specifically developed by Brookes et al. (2012) for the detection of sVCI. This thesis aimed to discuss about sVCI and to adapt BMET for use in subjects over 60 years old from Brazil. For this purpose, consecutive stages, comprising translation, evaluation by experts in linguistics and neuropsychology, assessment by representant of the target population, retro-translation and, finally, application to an older sample, have been performed. A pilot study has been carried out to analyze the applicability and to draw preliminary information about psychometric properties of the scale. In conclusion, BMET has revealed to be easy and rapid to administer, feasible to be applied by different categories of health practitioners and possibly adequate for primary health settings. Considering the current context of a fast population aging and a rise in the prevalence of conditions associated with late-life, such as neurocognitive disorders, it is relevant to detect sVCI in its early stages, so that therapeutic and secondary preventive strategies could be planned, which could impact on the quality of lives of older population.