Inducing Targeted Neuronal Injury Using a High-Power Laser in a Drosophila Larva

In This Article

Abstract

Source: Li, D., Li, F., et al. A Drosophila in vivo injury model for studying neuroregeneration in the peripheral and central nervous system. J. Vis. Exp. (2018)

This video demonstrates a method for inducing targeted neuronal injury in Drosophila larvae using a high-power laser. After positioning the larva under a microscope, a low-power laser scans for fluorescent neurons, and then the laser power is increased to induce localized damage. The injury is confirmed by increased fluorescence and a crater at the injury site.

Protocol

1. Two-photon Injury and Confocal Imaging

Microscope setup
NOTE: A confocal laser scanning microscope with a two-photon laser was used for this experiment, but other systems with an equivalent setup will also suffice. The two-photon laser (930 nm) was used for delivering injury and an Argon laser (488 nm) was used for confocal imaging of GFP.
1. At the beginning of each session, turn on the two-photon laser and/or the confocal laser(s) and the microscope. Then, open the imaging software.
2. For two-photon injury, set up the following parameters for imaging GFP with the two-photon laser at 930 nm (1,....

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Results

Larval axon analysis method, axon injury recovery, neuronal diagrams, microscopy results.
Figure 1: Da neuron axon regeneration in the periphery displays class specificity. (A and B) Schematic drawing showing the position of the larvae. (C) Schematic drawing of class III da neurons. (D) Axons of the class III da neurons ddaF, labeled with 19-12-Gal4,.......