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Synaptic Physiology Analysis Methods

Data generated during the experiment were analyzed in stages to identify and characterize synapses between distinct cell subclasses. Below is a workflow describing how this dataset was processed and analyzed; more detail can be found in Seeman, Campagnola et al. 2018.

 

Cell classification

Transgenic cell subclass was identified from the overlap of the fluorescent reporter of the cell and that of the recording pipette. Spiny and aspiny cells were defined from morphological analysis. Cells were annotated with a target layer during the experiment, and later a corrected layer determined from biocytin and DAPI staining.

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Synapse detection

Synapses were identified from manual inspection of the postsynaptic response aligned to the presynaptic spike. Excitatory and inhibitory connections could be observed in both voltage and current clamp at a holding potential that increased the driving force for each connection class. Weak synapses were detectable by averaging hundreds of postsynaptic responses.

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Synapse characterization

Resting-state synaptic strength, rise/decay kinetics, and depth of short-term plasticity were calculated from curve fits of postsynaptic responses.

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Quality control

Data passed through multiple quality control filters as part of the analysis workflow.

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