Drosophila Optogenetics: A Method to Manipulate Neuronal Circuits

In This Article

Abstract

Source: de Vries, S. E., Clandinin, T. Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster. J. Vis. Exp. (2013).

Optogenetics enables the use of light to manipulate neurons that are genetically engineered to express specific opsins–light-sensitive proteins whose light activation triggers a change in the state of the neuron. Here we describe an optogenetic approach in Drosophila using the opsin Channelrhodopsin2. The example protocol features an optogenetics assay set to study the neuronal circuitry behind the fly's escape behavior.

Protocol

This protocol is an excerpt from de Vries and Clandinin, Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster, J. Vis. Exp. (2013).

1. Generate Channelrhodopsin Flies

Cross UAS-ChR2 flies with the Gal4 driver of your choosing, we use G105-Gal4, which is expressed in Foma-1 neurons in the optic lobe.
To eliminate the possibility of a visual response to the blue light stimulation, both fly lines are in a w⁺norpA background.
End result: w⁺norpA;G105-Gal4/UAS-ChR2 +
After adult flies eclose, put selected females on fresh food, supplemented with 10....

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Results

LED cooling system setup, schematic, and components analysis; heat sink, lens, power supply diagram.
Figure 1: Experimental set-up showing the platform with the vertical holder and the four coolant hosing arms holding heat sinks with LED arrays affixed to them. (A) The set-up under ambient illumination. (B) The set-up when the LEDs are ill.......