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Show 87 Potential impact of this thesis work In utero electroporation (IUE) is a well-established method for gene transfection of astrocytes and neurons in the rodent brain (Chen and LoTurco, 2012; Nakahira and Yuasa, 2005; Tabata and Nakajima, 2001; Yoshida et al., 2010). In combination with the GCaMPcarrying plasmids described in this thesis work, IUE will enable exploration of astrocytic and neuronal intracellular calcium dynamics in rat models of neurophysiology and neuropathology without many of the limitations of dyes or viral-based methods (Gee et al., 2015; Reeves et al., 2011). IUE of GCaMPs is especially useful for monitoring astrocytic fine process dynamics and provides the opportunity to observe synaptic-related neuronalastrocytic interactions in the rat brain (Wilcox et al., 2015). Although not demonstrated in this work, in vivo imaging of the rat cortex following IUE should be straightforward. Hippocampus could possibly be imaged via techniques applied to mice, as well (Dombeck et al., 2010; Mizrahi et al., 2004). Thus, this toolset provides access to observation of unexplored astrocytic contributions to disease pathogenesis. Calcium imaging in the mammalian brain is most often performed in rats or mice. However, human behavior or disease may be more effectively modeled in other species (Cao et al., 2003; Shinya et al., 2011; Xu et al., 2013; Yang et al., 2013). For instance, studies of longevity-related disorders such as Alzheimer's may be more informative if performed in the naked mole-rat which has a lifespan of over 30 years, naturally develops some of the pathological findings observed in human Alzheimer's patients but does not develop behavioral symptoms of the disease (Edrey et al., 2013; Orr et al., 2015). IUE could likely be incorporated quickly into species in which transgenic technologies are immature or nonexistent. For imaging deeper structures in the intact mammalian brain, animals with smaller brains than mice such as the shrew, vole or mole could be imaged |
