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Activity-based automatic ROI generation (AARG) analysis of dendritic spine calcium transients reveals distance-dependent activity of voltage-gated calcium channels

dc.contributor.advisorDean, Camin Dr.
dc.contributor.authorGilbride, Charlie Jonathan
dc.titleActivity-based automatic ROI generation (AARG) analysis of dendritic spine calcium transients reveals distance-dependent activity of voltage-gated calcium channelsde
dc.contributor.refereeSchild, Detlev Prof. Dr. Dr.
dc.description.abstractengImaging synaptic activity in the form of calcium transients occurring in dendritic spines is advancing our understanding of nerve cell function beyond what can be achieved using electrophysiological methods alone. I have developed an analytical approach (which I name Activity-based Automatic Region of interest Generation (AARG) analysis) that facilitates the analysis of synaptic events simultaneously imaged across many dendritic spines. The approach automatically assigns regions of interest (ROIs) based on patterns of compartmentalized calcium transients in spines and generally emphasizes automation such that minimal user input is required. I demonstrate two ways in which this analysis can be applied to relevant questions in neuroscience. First, neurons are exposed to a recombinant neurotrophic compound – brain-derived neurotrophic factor (BDNF) – and then spontaneous spine calcium transients (SSCTs) are collected by washing magnesium out of the bath solution. Imaging SSCTs is required by AARG to automatically assign ROIs. With this approach, the effect of BDNF (or other compounds known to act on dendritic spines) can be analysed. Second, I have assessed the contribution of different voltage-gated calcium channels to SSCTs with a range of pharmacological agents. I find that specific blockers for channels mediating R-type and T-type currents contribute differently to SSCT depending upon distance of the current branch from the soma. These findings fit into an emerging picture of ion channel expression across the dendritic tree that appears well suited to support non-linear membrane potential fluctuations in dendrites. In addition to these imaging studies I have attempted to induce potentiation between pairs of synaptically connected neurons in dissociated hippocampal neurons. I was not able to induce stable potentiation under these conditions. Given the favourable experimental design I implemented (e.g. using perforated patch-clamp experiments) and based on a sound understanding of the relevant literature, signs of stable potentiation should have been more apparent. After thorough re-appraisal of the literature, I conclude that stable potentiation may require a more substantial input cooperativity than I initially
dc.contributor.coRefereeSchlüter, Oliver Dr. Dr.
dc.contributor.thirdRefereeRhee, Jeong Seop Dr.
dc.contributor.thirdRefereeDresbach, Thomas Prof. Dr.
dc.contributor.thirdRefereeHörner, Michael Prof. Dr.
dc.subject.engAutomatic ROI generationde
dc.subject.engDendritic spinesde
dc.subject.engCalcium transientsde
dc.subject.engSynaptic plasticityde
dc.affiliation.instituteGöttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)de
dc.subject.gokfullBiologie (PPN619462639)de

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