Studying the Excitability of Fluorescent Neurons using a Whole-Cell Patch Clamp

In This Article

Abstract

Source: Briz, C. G., et al., In Utero Electroporation Approaches to Study the Excitability of Neuronal Subpopulations and Single-cell Connectivity. J. Vis. Exp. (2017)

The video demonstrates a method to study the excitability of neuronal cells expressing fluorescent protein using a whole-cell patch current clamp. In the current-clamp mode, action potentials indicative of neuronal excitability are generated.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Electrophysiology

NOTE: The goal of this protocol is to obtain whole-cell current-clamp recordings from layer II/III pyramidal cell neurons identified visually by GFP expression in GFP-electroporated mouse brains (or any other fluorescent protein previously electroporated). Using this protocol, it is possible to study the effect of a genetic modification introduced by IUE on the electric properties of the neuron. The acquisition of specific firing modes is ....

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Results

Microscopy setup for GFP imaging; bright field and GFP merge; electrophysiology results.
Figure 1. Electrophysiology Settings and Example of a Firing Response. A. Photograph shows the electrophysiology setup used for patch clamp experiments in acute slices. The setup is included in a Faraday cage to eliminate noise, an.......