Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System
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Electroporation is an efficient method of transferring charged macromolecules into living cells in order to study their morphology, function, and connectivity within neuronal networks. Labeling cells with fluorophore-coupled macromolecules can be used to trace projections of whole neuronal ensembles, as well as the fine morphology of single cells. Here, we present a protocol to visualize pre- and postsynaptic components of a sensory relay synapse in the brain, using the olfactory system of Xenopus laevis tadpoles as a model. We apply bulk electroporation to trace projections of receptor neurons from the nose to the brain, and single cell electroporation to visualize the morphology of their synaptic target cells, the mitral-tufted cells. Labeling the receptor neurons with a calcium-sensitive dye allows us to record stimulus-induced presynaptic input to the dendrites of the postsynaptic cells via functional calcium imaging.
Key wordsElectroporation Neuronal tracing Single cell morphology Fluorescent dyes Calcium imaging In vivo imaging Xenopus laevis Olfactory system Olfactory receptor neurons Mitral-tufted cells
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