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Vasopressin anatomy of the mouse brainRood, Benjamin D 01 January 2010 (has links)
The nine amino acid peptide vasopressin acts as a neurohormone in the periphery and a neurotransmitter/neuromodulator in the central nervous system. Historically, research on vasopressin neurons and their projections to the pituitary has helped lay the groundwork for our understanding of peptidergic neurotransmission. Currently, our research on central vasopressin projections is driving a revolution in our understanding of social behavior. Vasopressin affects a number of social behaviors from social memory to aggression to affiliative behavior, such as pair-bonding. Further, with the addition of more and more transgenic mouse models of disease states, anxiety and depression related disorders, and social behavior dysfunction, it is important now more than ever to have a clear knowledge of the mouse vasopressin system, which derives from a number of distinct nuclei within the brain. Here, I map out vasopressin immunoreactivity in the mouse brain, and delineate the subset of brain regions with gonadal steroid hormone-dependent vasopressin immunoreactivity. Such projections are thought to derive from the bed nuclei of the stria terminalis and medial amygdala in the telencephalon. Finally, based on data from mice with lesions of the suprachiasmatic nucleus, I outline the subset of regions that likely receive vasopressin from this source. Our research on the anatomy of the vasopressin system of mice and our attempts to delineate the site of origin of the many vasopressin fibers found throughout the brain suggest that a significant amount of the vasopressin innervation deriving from cells in the bed nuclei of the stria terminalis and medial amygdala project to areas in the midbrain involved in serotonin and dopamine transmission, such as the dorsal raphe and ventral tegmental area. These transmitter systems play a crucial role in the control of anxiety and depression levels as well as motivated behavior and emotional regulation. Our results strongly suggest that a direct link exists between these systems, and future plans include an examination of this possibility. It is our hope that this work will further our understanding of the role of vasopressin and other transmitter systems in the regulation of social behaviors.
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Engineered Micro and Nanotopography for Neuron and Glial Guidance after Spinal Cord InjuryJohnson, Christopher David Landis 09 August 2018 (has links)
<p> Spinal cord injury presents many barriers to regeneration that have yet to be solved. Guidance conduits are a promising approach that allow regenerating tissue to bypass or traverse the injury site. Peripheral nerve autografts into the spinal cord have guided neurons and glia around an injury site and resulted in some functional recovery [1]. Synthetic guidance conduits made from electrospun fibers have shown an ability to guide robust regeneration after spinal cord injury [2]–[4]. The surface topography of the electrospun fiber scaffolds is important for cell guidance, and can be modified by changing the fiber diameter, fiber alignment, and fiber surface nanotopography, among others. While the effects of these fiber physical characteristic on neuron guidance has been determined, it remains unknown how fiber diameter and surface nanotopography affect astrocytes - the most abundant cells in the CNS. Astrocytes maintain homeostasis in the healthy CNS and provide guidance and protection to neurons after injury. As a result, astrocytes should be considered an important therapeutic target for biomaterial approaches. The current thesis studied the effects of surface topography on astrocyte morphology and astrocyte phenotype. </p><p> First, the astrocyte response to fiber surface nanotopography was tested. Astrocytes were cultured on three engineered surface topographies - smooth, pitted, or divoted fiber surfaces. Astrocytes isolated from the spinal cord were compared to astrocytes isolated from the cortex, to determine if there was a different response from astrocytes selected from different anatomical locations. The astrocyte morphological response was measured with immunocytochemistry, while the GFAP and vinculin protein expression was measured using Western blot techniques. The findings suggest that smooth fiber surfaces allowed for the longest cortical astrocyte extension after 4 days in culture, while the pits and divots reduced astrocyte elongation. The spinal cord astrocytes exhibited smaller elongation ratios over the times tested, compared to the cortical astrocytes, and did not have significantly different responses to the fiber surfaces tested. Since astrocyte elongation is correlated with neurite outgrowth, these data shows that the smooth fibers provide the greatest astrocyte elongation. The GFAP and vinculin expression were not significantly different among any of the surfaces or cell types tested. </p><p> Second, the astrocyte response to fiber diameter was tested. Astrocytes were cultured on large (800nm) diameter, small (386nm) diameter, and film substrates, and the astrocyte morphology was analyzed over the first four days in culture. Astrocytes became significantly more elongated on the large fibers as early as two hours after culture, and remained significantly more elongated after four days. The effect of fiber diameter on GLT-1, GLAST, and GFAP expression were examined by western blot. GLT-1 was significantly increased on the fiber substrates, compared to the film, but GLT-1 expression was not dependent on fiber diameter. A glutamate excitotoxicity assay was performed on an astrocyte/neuron coculture to determine if the increased astrocyte GLT-1 expression improved neuron protection. The findings suggest that fibrous surfaces improved cell survival, however, the differences were not statistically significant from the film control. Next neuron outgrowth was observed on the astrocyte surfaces prepared on each of the three topographies - small fiber, large fiber, and film. Neurons were cultured on the astrocyte/fiber surfaces and compared to an astrocyte/film control. The results suggest that after 4 days in culture, the astrocytes on the large fibers induced longer more directed growth, while the astrocytes on the small fibers induced comparatively shorter neurite growth with more neurite branching. </p><p> Third, a novel biomaterial scaffold was prepared by incorporating superparamagnetic iron oxide nanoparticles (SPIONs) into the electrospinning solution. The purpose was to prepare an injectable electrospun fiber scaffold that could be magnetically oriented <i>in situ</i>. The surface topography was carefully controlled. The effect of incorporating increased amounts of SPIONs on the fiber diameter, fiber surface nanotopography, and fiber alignment was measured, and compared against the previous findings in this thesis and against a similar scaffold that has been successful <i>in vivo</i>. The effect of the increasing SPION concentration on the fiber magnetization and on the speed of fiber alignment in viscous solutions was measured. From these data, the 6% SPION concentration exhibited the fastest fiber realignment times without significant changes to the fiber topography. As a result the 6% SPION fibers were tested in vitro by orienting the fibers to provide an aligned fiber scaffold within either a collagen/Matrigel or fibrin/Matrigel hydrogel. (Abstract shortened by ProQuest.)</p><p>
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