Amygdala, especially the lateral central nucleus (CeL), plays a critical role in mediating anxious temperament (AT). However, the cellular, molecular, and functional properties of CeL neurons that underlie AT pathogenesis remain to be explored. The Cellular Neurobiology Core will provide standard technical services to characterize the gene (protein) expression in CeL neurons at the light and electron microscopic levels in qualitative and quantitative manners. We will reprogram induced pluripotent stem cells (iPSCs) from skin biopsy tissues of monkey and human subjects with high and low AT using the state-of-the-art nonintegration technology. These IPSCs will be directed to neuronal subtypes, including the DARPP32- expressing medium spiny GABA neurons that resemble those in the CeL using our established protocols. The gene expression, transmitter release and electrophysiological properties of these neurons will be evaluated by immunocytochemistry, HPLC, and whole cell patch clamping analysis. These cellular properties will be cross verified between cultured neurons and those in the monkey brain and compared between high and low AT. By providing these services and close collaboration with individual projects, the core will generate iPSCs from high and low AT monkeys and human individuals for the first time. These cell lines will become valuable resources for and be made available to investigators across the Conte centers. Information gained from the cellular analyses will enable the discovery of cellular/ functional substrate that underlies AT development and bridge the monkey model to human disorders.
The Cellular Neurobiology Core will provide essential technical services to reveal cellular, molecular, and functional changes in neurons that are responsible for AT development for the first time, especially those from AT human individuals that are otherwise inaccessible.
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