The dorsal lateral geniculate nucleus (dLGN) has become an important model for studying many aspects of visual system development. To date, studies have focused on the development of retinal projections and the role of activity in shaping the pattern of synaptic connections made with thalamocortical relay cells. By contrast, little is known about relay cells and the factors that regulate the growth and establishment of their dendritic architecture. In many systems, such growth seems consistent with the synaptotrophic hypothesis which states that synapse formation and dendritic growth work in a concerted fashion such that afferent input and the establishment of functional synapses are needed to shape the maturation of dendritic arbors. To address this, we characterized the development of relay cells in the dLGN of wild-type (WT) mouse. By adopting a loss of function approach, we assessed the manner in which growth and maturation of relay cells were affected by retinal innervation. For this, we made use of the math-null (math5-/-) mouse in which progenitors fail to differentiate into retinal ganglion cells (RGCs), and exhibit a >95% cell loss. Anterograde labeling of RGC axons with cholera toxin subunit B (CTB), immunolabeling of RGC-specific presynaptic machinery in dLGN (e.g. vesicular glutamate transporter 2), and ultrastructural analysis at the electron microscopy level demonstrated that the dLGN is devoid of retinal innervation. We examined the functional and morphological characteristics of relay cells in WT and math5-nulls during early postnatal life by conducting in vitro whole cell recordings in slices containing dLGN. Individual relay cells were labeled by intracellular injection of biocytin, and imaged by confocal microscopy to obtain the 3-D reconstructions of their dendritic trees. Morphometric analysis revealed that relay cells in WT undergo two growth spurts: an early one where cell class specification and dendritic complexity are established and a later one marked by an increase in dendritic field and length. Following the third week, relay cells growth was stabilized. In math5-nulls, relay cells maintained their morphological identity whereby cells could be classified in three groups (Y: spherical, X: bi-conical, W: hemi-spherical). However, the dLGN was highly reduced in size, and relay cells showed disrupted growth spurts. Relay cells had smaller somata and exhibited fluctuations in dendritic complexity and field extent compared to age-matched WTs. Exuberance in dendritic branching was noted in week 2, and by week 5, relay cells had significantly smaller surface area resulting from a loss of dendritic segments and a reduction in dendritic field extent. Control experiments using RT-PCR revealed that these changes were not due to the loss of math5 in the dLGN. Whole cell recordings and voltage responses to square wave current pulses showed that math5-nulls possess the full compliment of intrinsic membrane properties, such as relay cells displayed both burst and tonic firing modes. A cross of the math5-null with a transgenic mouse that expresses GFP in layer VI cortical neurons revealed a dense plexus of corticogeniculate terminals throughout the mature dLGN. However, the rate of corticogeniculate innervation was highly accelerated and was complete a week earlier than WT. Electric stimulation of cortical axons revealed that synapses are functional and responses were indistinguishable from WT. Taken altogether, these observations suggest that retinal innervation plays an important trophic role in the maturation of dLGN and is necessary for the continued maintenance of relay cells’ structural integrity. However, the general form and function of relay cells seem largely unaffected by the loss of retinal innervation.

Chez diverses espèces animales, les informations sensorielles peuvent déclencher la locomotion. Ceci nécessite l’intégration des informations sensorielles par le système nerveux central. Chez la lamproie, les réseaux locomoteurs spinaux sont activés et contrôlés par les cellules réticulospinales (RS), système descendant le plus important. Ces cellules reçoivent des informations variées provenant notamment de la périphérie. Une fois activées par une brève stimulation cutanée d’intensité suffisante, les cellules RS produisent des dépolarisations soutenues de durées variées impliquant des propriétés intrinsèques calcium-dépendantes et associées à l’induction de la nage de fuite. Au cours de ce doctorat, nous avons voulu savoir si les afférences synaptiques ont une influence sur la durée des dépolarisations soutenues et si l’ensemble des cellules RS partagent des propriétés d’intégration similaires, impliquant possiblement les réserves de calcium internes. Dans un premier temps, nous montrons pour la première fois qu’en plus de dépendre des propriétés intrinsèques des cellules réticulospinales, les dépolarisations soutenues dépendent des afférences excitatrices glutamatergiques, incluant les afférences spinales, pour perdurer pendant de longues périodes de temps. Les afférences cutanées ne participent pas au maintien des dépolarisations soutenues et les afférences inhibitrices glycinergique et GABAergiques ne sont pas suffisantes pour les arrêter. Dans un deuxième temps, nous montrons que suite à une stimulation cutanée, l’ensemble des cellules RS localisées dans les quatre noyaux réticulés possèdent un patron d’activation similaire et elles peuvent toutes produire des dépolarisations soutenues dont le maintien ne dépend pas des réserves de calcium internes. Enfin, les résultats obtenus durant ce doctorat ont permis de mieux comprendre les mécanismes cellulaires par lesquels l’ensemble des cellules RS intègrent une brève information sensorielle et la transforment en une réponse soutenue associée à une commande motrice. / In various animal species, sensory information can initiate locomotion. This relies on the integration of sensory inputs by the central nervous system. In lampreys, the spinal locomotor networks are activated and controlled by the reticulospinal cells (RS) which constitute the main descending system. In turn, RS cells receive information coming from various synaptic inputs such as the sensory afferents. Once activated by a brief cutaneous stimulation of sufficient strength, RS cells display sustained depolarizations of various durations that rely on calcium-dependant intrinsic properties and lead to the onset of escape swimming. During the course of this Ph.D, we aimed at determining whether synaptic inputs can modulate the duration of the sustained depolarizations and if the different populations of RS cells share the same integrative properties, possibly involving the internal calcium stores. First, our results show for the first time that excitatory glutamatergic inputs, including ascending spinal feedback, contribute to prolong the sustained depolarizations for long periods of time. Cutaneous inputs do not contribute to maintain the sustained depolarizations and inhibitory glycinergic and GABAergic inputs are not sufficient to stop them. Second, we show that in response to cutaneous stimulation, the RS located in the four reticular nuclei display a similar activation pattern and can all produce sustained depolarizations which do not depend on internal calcium release to be maintained. Finally, the results obtained during this Ph.D allowed us to better understand the cellular mechanisms by which the RS cells integrate and transform a brief sensory information into a sustained response associated with a motor command.

Chez diverses espèces animales, les informations sensorielles peuvent déclencher la locomotion. Ceci nécessite l’intégration des informations sensorielles par le système nerveux central. Chez la lamproie, les réseaux locomoteurs spinaux sont activés et contrôlés par les cellules réticulospinales (RS), système descendant le plus important. Ces cellules reçoivent des informations variées provenant notamment de la périphérie. Une fois activées par une brève stimulation cutanée d’intensité suffisante, les cellules RS produisent des dépolarisations soutenues de durées variées impliquant des propriétés intrinsèques calcium-dépendantes et associées à l’induction de la nage de fuite. Au cours de ce doctorat, nous avons voulu savoir si les afférences synaptiques ont une influence sur la durée des dépolarisations soutenues et si l’ensemble des cellules RS partagent des propriétés d’intégration similaires, impliquant possiblement les réserves de calcium internes. Dans un premier temps, nous montrons pour la première fois qu’en plus de dépendre des propriétés intrinsèques des cellules réticulospinales, les dépolarisations soutenues dépendent des afférences excitatrices glutamatergiques, incluant les afférences spinales, pour perdurer pendant de longues périodes de temps. Les afférences cutanées ne participent pas au maintien des dépolarisations soutenues et les afférences inhibitrices glycinergique et GABAergiques ne sont pas suffisantes pour les arrêter. Dans un deuxième temps, nous montrons que suite à une stimulation cutanée, l’ensemble des cellules RS localisées dans les quatre noyaux réticulés possèdent un patron d’activation similaire et elles peuvent toutes produire des dépolarisations soutenues dont le maintien ne dépend pas des réserves de calcium internes. Enfin, les résultats obtenus durant ce doctorat ont permis de mieux comprendre les mécanismes cellulaires par lesquels l’ensemble des cellules RS intègrent une brève information sensorielle et la transforment en une réponse soutenue associée à une commande motrice. / In various animal species, sensory information can initiate locomotion. This relies on the integration of sensory inputs by the central nervous system. In lampreys, the spinal locomotor networks are activated and controlled by the reticulospinal cells (RS) which constitute the main descending system. In turn, RS cells receive information coming from various synaptic inputs such as the sensory afferents. Once activated by a brief cutaneous stimulation of sufficient strength, RS cells display sustained depolarizations of various durations that rely on calcium-dependant intrinsic properties and lead to the onset of escape swimming. During the course of this Ph.D, we aimed at determining whether synaptic inputs can modulate the duration of the sustained depolarizations and if the different populations of RS cells share the same integrative properties, possibly involving the internal calcium stores. First, our results show for the first time that excitatory glutamatergic inputs, including ascending spinal feedback, contribute to prolong the sustained depolarizations for long periods of time. Cutaneous inputs do not contribute to maintain the sustained depolarizations and inhibitory glycinergic and GABAergic inputs are not sufficient to stop them. Second, we show that in response to cutaneous stimulation, the RS located in the four reticular nuclei display a similar activation pattern and can all produce sustained depolarizations which do not depend on internal calcium release to be maintained. Finally, the results obtained during this Ph.D allowed us to better understand the cellular mechanisms by which the RS cells integrate and transform a brief sensory information into a sustained response associated with a motor command.