Cell Ablation Techniques for the Larval Drosophila Neuromuscular System

Abstract

Tissue development requires local and long-distance communication between cells. Cell ablation experiments have provided critical insights into the functions of specific cell types and the tissue surrounding the dead cells. In the Drosophila neuromuscular system, ablation of motor neurons and muscles has revealed the roles of the ablated cells in axon pathfinding and circuit wiring. For example, when muscles are denervated due to laser ablation of their motor neuron inputs, they receive ectopic innervation from neighboring motor neurons. Here, we describe two methods of specific cell ablation. The first is a genetic ablation approach that uses GAL4 (ideally expressed in a small subset of cells) to drive expression of cell death genes reaper and head involution defective. The second method relies on reactive oxygen species produced by light activation of the Arabidopsis-derived Singlet Oxygen Generator, miniSOG2, expressed in a subset of cells. For the latter, the precision stems from both the GAL4 and the restricting of the blue-light stimulation area.