Monitoring Membrane Excitability in Drosophila Expressing Modified Shaker Constructs

Abstract

The Drosophila neuromuscular junction (NMJ) ranks as one of the preeminent model systems for studying synaptic development, function, and plasticity. This protocol describes the use of the two-electrode voltage clamp (TEVC) to examine potassium (K⁺) currents mediated by voltage-gated ion channels, and gives several genetic and pharmacological methods that are used to study the currents. Drosophila larval muscle fibers possess three major K⁺ currents. One of these, a fast voltage-activating and inactivating I_A current, is mediated by the Shaker channel. The Shaker channel is characterized by its sensitivity to the drug 4-aminopyridine (4-AP). Two useful transgenic tools for altering membrane excitability have been developed by making specific modifications of the Shaker channel; their use is described here.