An assay for visual learning in individual Drosophila larvae

Vision is a major sensory modality in Drosophila behavior, with more than one-half of the Drosophila brain devoted to visual processing. The mechanisms of vision in Drosophila can now be studied in individuals and in populations of flies by using various paradigms. Simple strategies for conducting visual perception and learning studies consist of individual studies performed on single flies on solid supports (larvae on agar or adults in a T-maze) using a light/dark association paradigm. These approaches are quite easy to implement but are fairly limited in their ability to address questions of visual perception. Nevertheless, the simpler approaches treating vision in one dimension (light, dark) do provide effective paradigms for genetic analysis. This article describes a protocol for larval visual learning. Larvae are transferred back and forth between well-lit or dark agarose plates that either do or do not contain fructose (which is appetitive) as the unconditioned stimulus (US). Such appetitive larval visual memory is tested by tallying the time spent in light or dark quadrants of a plate (without a US).