Chapter title |
Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System
|
---|---|
Chapter number | 15 |
Book title |
Xenopus
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8784-9_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8783-2, 978-1-4939-8784-9
|
Authors |
Weiss, Lukas, Offner, Thomas, Hassenklöver, Thomas, Manzini, Ivan, Lukas Weiss, Thomas Offner, Thomas Hassenklöver, Ivan Manzini |
Abstract |
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. |
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