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Title: Imaging neural activity in vitro and in vivo using genetically-encoded calcium indicators
Author: Walker, Alison Sarah
ISNI:       0000 0004 5357 4227
Awarding Body: King's College London (University of London)
Current Institution: King's College London (University of London)
Date of Award: 2014
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The projects in this thesis use genetically-encoded calcium indicators (GECIs) to investigate neural activity both in vitro, in dissociated hippocampal neurons, and in vivo, in the visual system of the larval zebrafish. We first characterised the properties of established and newly-generated GECIs in dissociated hippocampal neurons. Our first question dealt with understanding how quantal excitatory activity is distributed across the dendritic tree of dissociated hippocampal neurons. In neurons expressing GCaMP3 we recorded small NMDA receptor-dependent calcium transients, in the presence of tetrodotoxin. These were localised to dendritic spines and highly heterogeneous in terms of amplitude and frequency, both at single synapses and across the dendritic tree. Precise mapping of quantal synaptic events onto dendritic arbours of individual neurons determined that the amplitude of calcium transients correlated with the location of the spine within a branch, with larger calcium transients found closer to branch ends. One aim of this thesis was to investigate how this spatial distribution of activity arises. A second question was to understand how single motion-sensitive neurons in the retinotectal system of the larval zebrafish develop their functional properties? The function of single retinal ganglion cells labelled with SyGCaMP3, or single tectal cells labelled with RGECO was probed by playing a drifting bar visual presentation to the same awake larvae at different timepoints during their development (3-7 days post fertilisation). As larval zebrafish can see prior to the full maturation of their visual circuitry, our approach gives insight into how ongoing circuit maturation impacts on the function output of single neurons. We discovered that some classes of motion-sensitive neurons appear functionally invariant during development, suggesting insensitivity to ongoing circuit refinement, while others classes mature through a period of functional refinement.
Supervisor: Meyer, Martin Patrick; Burrone, Juan Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available