Magnocellular neurosecretory cells (MNCs) are neuroendocrine cells with somata located in the hypothalamus and nerve terminals in the posterior pituitary. They receive neuronal inputs from the brain and release vasopressin and oxytocin into the blood to regulate many important functions such as water balance, lactation, and parturition. The process of hormone release depends on Ca2+ influx mediated by voltage-gated Ca2+ channels (VGCCs) on the plasma membranes of neuroendocrine cells. To better understand the cellular and molecular components that are involved in regulating secretory vesicle exocytosis, this thesis work was conducted to investigate the expression and function of different subtypes of VGCCs in MNCs and pituicytes (the glial cells surrounding MNC nerve terminals).<p> Molecular biology, immunohistochemistry and cellular biology were used to detect expression and alternative splicing of different VGCC subtypes in MNCs, neurons, and pituicytes. First, the presence of CaV2.2 and CaV2.3 channels were detected on the pituicytes in situ. When the pituicytes were isolated and cultured for 14 days, more VGCC subtypes were expressed including CaV1.2 channels. Regulation of VGCC expression was measured in normal and dehydrated rats, and CaV1.2 channels were found to be selectively up-regulated in pituicytes after 24 hours of dehydration.<p> Second, two splice variants of CaV2.1 channels (CaV2.1Ä1 and Ä2) that lack a large portion of the synprint (synaptic protein interaction) site were detected in the rat brain. To determine whether the splice variants were expressed in MNCs, we did immunocytochemistry using two antibodies (the selective and the inclusive antibody) that recognized the carboxyl-terminus of channels and the synprint site, respectively, in different cell types. We found that vasopressin MNCs, but not the oxytocin MNCs, and one type of endocrine cell (the melanotropes of the pituitary gland) expressed the synprint site deleted variants, whereas the hippocampal neurons mainly expressed the full-length isoform. The splice variants were properly distributed on the plasma membrane of the somata and nerve terminals of the MNCs, suggesting the synprint site is not essential for CaV2.1 channel targeting into the nerve terminals of neuroendocrine cells.<p> Third, expression and distribution of CaV2.2 channels were studied in the MNCs. All CaV2.2 isoforms we detected contained the full-length synprint site. To test the importance of the CASK/Mint1 binding site for CaV2.2 channel targeting, we over-expressed a peptide that inhibits the interaction between CaV2.2 channels and CASK/Mint1 in differentiated PC12 cells (a neuroendocrine cell line). We found that the distribution of CaV2.2 channels in the growth cones of PC12 cells were significantly decreased, suggesting that the CASK/Mint1 interaction is important for CaV2.2 channel targeting into the neuroendocrine terminals.<p> In conclusion, these results provide insights of VGCC expression in neuroendocrine cells, and also give rise to a better understanding of the molecular components that are involved in forming the exocytotic machinery in these cells.

Acute brain cell swelling (cytotoxic edema) can occur in the first minutes of stroke, presumably as a result of brain cells taking up water. In extreme hypo-osmotic situations such as excessive water-loading by patients, uptake by brain cells can expand the brain, causing seizures. But is ischemic brain cell swelling the same as hypo-osmotic swelling? Water can passively diffuse across the plasma membrane. However the presence of water channels termed aquaporins (AQP) facilitates passive water diffusion by 10-100 times. Unlike astrocytes, there is no evidence of water channels on neuronal plasma membrane. However, there is still much debate about which cells (neurons or astrocytes) swell during over-hydration or during stroke and if neurons and astrocytes can volume-regulate during osmotic stress. The purpose of this study was to examine and compare the volume responses of PyNs and magnocellular neuroendocrine cells (MNCs) to acute osmotic challenge and to OGD. We examined MNCs because they are intrinsically osmosensitive to small changes (2-3 mOsm) of plasma osmolality. We also examined if the same neurons behave similarly in brain slices or when dissociated and if they respond differently to acute osmotic stress and stroke-like stress. Our results indicate that during acute osmotic stress (±40 mOsm) half of dissociated PyNs and MNCs tended to show appropriate responses. MNCs in brain slices showed similar responses to when they were dissociated, while brain slice PyNs were less responsive than when dissociated. Exposure to OGD resulted in obvious differences between the two types of in vitro preparations. Dissociated PyNs and MNCs showed no consistency in their volume responses to 10 minutes of OGD. Dissociated neurons swelled, shrunk or were unchanged in about equal numbers. In contrast, brain slice PyNs underwent profound swelling whereas, brain slice MNCs showed minor volume decreases. We conclude that about half of our dissociated neurons were too variable and unpredictable in their osmotic volume responses to be useful for osmotic studies. They also were too resistant to stroke-like stress to be good models for ischemia. Brain slice neurons were similar in their osmotic responses to dissociated neurons but proved to have consistent and predictable responses to stroke-like stress. / Thesis (Master, Neuroscience Studies) -- Queen's University, 2011-02-07 17:55:08.333

Cette thèse avait pour principal objectif d’étudier la dissociation du système magnocellulaire et parvocellulaire en testant l’évolution de ces systèmes dans le vieillissement normal et la réhabilitation à l’aide de lunettes à filtre jaune. Pour cela deux paradigmes expérimentaux dont les caractéristiques des stimuli permettaient de biaiser le traitement visuel vers l’un ou l’autre des deux systèmes et dont les tâches impliquaient un niveau de traitement cognitif plus ou moins élaboré ont été utilisés. A l’aide d’une tâche de discrimination de luminance, les travaux ont mis en évidence l’apparition d’un déficit parvocellulaire massif dans le vieillissement ainsi qu’un déficit magnocellulaire de moindre ampleur au début du vieillissement mais qui s’accentuait rapidement au-delà de 75 ans. Dans une tâche de catégorisation d’objets, la présence du déficit magnocellulaire avec l’âge et le grand âge a été corroborée mais aucune répercussion du déficit parvocellulaire mis en évidence à un niveau de traitement plus précoce n’a été observée dans cette tâche. Concernant la réhabilitation, seul le groupe de participants très âgé (>75 ans) a vu ses performances améliorées par le filtre jaune dans la condition impliquant spécifiquement le système magnocellulaire à l’épreuve de discrimination de luminance. Dans l’épreuve de catégorisation, les deux groupes de participants âgés et très âgés ont montré une diminution significative de leur temps de réponse avec le filtre jaune accompagnée d’une amélioration de la précision de catégorisation pour le groupe très âgé, spécifiquement dans la condition mettant en jeu le système magnocellulaire.L’ensemble des résultats confirment donc la dissociation neuropsychologique entre système magnocellulaire et parvocellulaire : ils peuvent être sollicités de façon privilégiée chez l’adulte jeune en faisant varier à la fois plusieurs caractéristiques de la stimulation et la tâche ; ils ne vieillissent pas au même rythme et l’un d’entre eux peut être spécifiquement réhabilité / The thesis aimed at studying the dissociation between the magnocellular and the parvocellular system by testing their evolution in normal aging and their rehabilitation with yellow filter glasses. Two experimental paradigms whose specific stimuli characteristics and task allow to bias visual processing toward one or the other system and which involved a low or an elaborated cognitive processing were used.In a luminance discrimination task, the results showed a massive parvocellular and a lower magnocellular deficit at the beginning of aging, but the magnocellular deficit increased rapidly beyond 75 years old. In an object categorization task, the presence of magnocellular deficit with age and very old age was corroborated but there was no parvocellular deficit suggesting that the parvocellular deficit found at earlier stage of processing had no consequence for higher-level processes.Regarding rehabilitation, yellow filter improved sensitivity in the discrimination of luminance specifically in the condition involving the magnocellular system and for the very old group of participants (> 75 years old) only. In the categorization task, yellow filter significantly decreased response time specifically in the condition isolating the magnocellular system for both old groups and increased the accuracy for the very-old group.Taken together, the results confirm the dissociation between the magnocellular and parvocellular systems. Magnocellular system undergoes age-related changes at later stage of ageing than parvocellular system and it could be rehabilitated with yellow filter

No description available.

L’expression faciale de peur constitue un important vecteur d’information sociale mais aussi environnementale. En condition naturelle, les visages apeurés apparaissent principalement dans notre champ visuel périphérique. Cependant, les mécanismes cérébraux qui sous-tendent la perception de l’expression faciale de peur en périphérie restent largement méconnus. Nous avons démontré, grâce à des études comportementales, des enregistrements magnétoencéphalographiques et intracrâniens, que la perception de l’expression faciale de peur est efficace en grande périphérie. La perception de la peur en périphérie génère une réponse rapide de l’amygdale et du cortex frontal, mais également une réponse plus tardive dans les aires visuelles occipitales et temporales ventrales. Le contrôle attentionnel est capable d’inhiber la réponse précoce à l’expression de peur, mais également d’augmenter les activités postérieures plus tardives liées à la perception des visages. Nos résultats montrent non seulement que les réseaux impliqués dans la perception de la peur sont adaptés à la vision périphérique, mais ils mettent également en avant une nouvelle forme d’investigation des mécanismes de traitement de l’expression faciale, pouvant conduire à une meilleure compréhension des mécanismes de traitement des messages sociaux dans des situations plus écologiques. / Facial expression of fear is an important vector of social and environmental information. In natural conditions, the frightened faces appear mainly in our peripheral visual field. However, the brain mechanisms underlying perception of fear in the periphery remain largely unknown. We have demonstrated, through behavioral, magnetoencephalographic and intracranial studies that the perception of fear facial expression is efficient in large peripheral visual field. Fear perception in the periphery produces an early response in the amygdala and the frontal cortex, and a later response in the occipital and infero-temporal visual areas. Attentional control is able to inhibit the early response to fear expression and to increase the later temporo-occipital activities linked to face perception. Our results show that networks involved in fear perception are adapted to the peripheral vision. Moreover, they validate a new form of investigation of facial expression processing, which may lead to a better understanding of how we process social messages in more ecological situations.

L’objectif de ce travail de thèse était d’étudier l’implication des systèmes visuels magnocellulaire et parvocellulaire dans le traitement de langage écrit, spécialement dans l’identification des mots. Quatre études expérimentales ont été réalisées afin de documenter le rôle respectif de ces deux systèmes pour différentes composantes de la structure du mot écrit, la lettre (Etude 1), la longueur du mot (Etude 2) et l’enveloppe du mot (Etude 3), ainsi que le voisinage orthographique (Etude 4). La dissociation des deux sous-systèmes visuels se fondait sur leurs caractéristiques fonctionnelles spécifiques : sensibilité aux fréquences spatiales basses et au contraste de luminance pour le système magnocellulaire ; sensibilité aux fréquences spatiales moyennes et élevées et au contraste chromatique pour le système parvocellulaire. Les participants étaient des adultes jeunes normolecteurs et des enfants de 10-11 ans, lecteurs novices, afin d’envisager l’implication de chaque système visuel à deux étapes de leur développement : maturité chez l’adulte jeune ; en cours de maturation chez l’enfant. Les résultats ont confirmé le rôle prépondérant des traitements parvocellulaires, ce qui corroborait le privilège accordé au traitement des lettres et traits des lettres dans la reconnaissance visuelle des mots. Le traitement de la longueur du mot s’est en revanche révélé être une dimension sélectivement magnocellulaire. L’ensemble des résultats est discuté dans le cadre de l’approche coarse-to-fine. / The main objective was to study the implication of the magnocellular and the parvocellular visual systems in written language processing, specifically in word identification. Four studies were carried out to document the respective roles of these two systems, for different components of the written word structure, the letter (Study 1), the word length (Study 2), the word shape (Study 3) and the orthographic neighborhood (Study 4). The dissociation of the two visual systems was based on their specific functional characteristics: sensitivity to low spatial frequencies and luminance contrast for the magnocellular system; and sensitivity to medium and high spatial frequencies and chromatic contrast for the parvocellular system. The participants were young adult normal readers and 10-11 years-old children, novice readers, in order to consider the involvement of each visual system at two stages of their development: mature in young adults while still maturing in the children. The results confirmed the prominent role of parvocellular processing, which was consistent with the privilege accorded to the processing of letters and letters features in visual word recognition. On the other hand, word length processing has been shown to be a selectively magnocellular dimension. All results are discussed in the framework of the coarse-to-fine approach.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

The magnocellular neurosecretory cells (MNCs) of the hypothalamus project axons from the supraoptic nucleus to the posterior pituitary gland, where they secrete either oxytocin or vasopressin into the circulation. Oxytocin is important for delivery at birth and is essential for milk ejection during suckling. Vasopressin primarily promotes water reabsorption in the kidney to maintain body fluid balance. The profile of oxytocin and vasopressin secretion is principally determined by the pattern of action potentials initiated at the cell bodies in the hypothalamus. MNCs principally secrete hormones from terminals in the pituitary, but the secretion also occurs from their dendrites in the supraoptic nucleus, where they diffuse and affect the neighbouring cells. Mechanisms controlling the oxytocin and vasopressin secretion from MNCs have been extensively studied over the last decades and it is assumed that the relationship between Ca2+ signalling, secretion from dendrites, and the firing patterns is essential in understanding the magnocellular neurosecretory system. In this project, we combine mathematical analysis and experimental measurements of Ca2+ activity of MNCs of transgenic rats expressing an arginine vasopressin-enhanced green fluorescent protein (AVP-eGFP) or oxytocin-monomeric red fluorescent...

After over a decade of its discovery, the Electroretinogram (ERG) still remains the objective tool that is conventionally used in assessment of retinal function in health and disease. Although there is ongoing research in developing ERG- recording techniques, interpretation and clinical applications, there is still a limited understanding on how each photoreceptor class contribute to the ERG waveform and their role and/or susceptibilities in various retinal diseases still remains unclear. Another limitation with currently used conventional testing protocols in a clinical setting is the requirement of an adaptation period which is time-consuming. Furthermore, the ERG responses derived in this manner are recorded under different stimulus conditions, thus, making comparison of these signals difficult. To address these issues and develop a new testing method, we employed silent substitution paradigm in obtaining cone- and rod- isolating ERGs using sine- and square- wave temporal profiles. The ERGs achieved in this manner were shown to be photoreceptor-selective. Furthermore, these responses did not only provide the functional index of photoreceptors but their contributions to their successive postreceptoral pathways. We believe that the substitution stimuli used in this thesis could be a valuable tool in functional assessment of individual photoreceptor classes in normal and pathological conditions. Furthermore, we speculate that this method of cone/rod activity isolation could possibly be used in developing faster and efficient photoreceptor-selective testing protocols without the need of adaptation. / Bradford School of Optometry and Vision Sciences