title: Using Neurogenetics and the Warmth-Gated Ion Channel TRPA1 to Study the Neural Basis of Behavior in Drosophila.
abstract: Here we describe a set of straightforward laboratory exercises that integrate the study of genetics, neuroanatomy, cellular physiology and animal behavior. We use genetic tools in Drosophila for visualizing and remotely activating ensembles of neurons with heat pulses. First, we show how to examine the anatomy of several neuronal populations using genetically encoded green fluorescent protein. Next we demonstrate how to use the warmth gated Drosophila TRPA1 (dTRPA1) cation channel to remotely activate neural circuits in flies. To demonstrate the cellular effects of dTRPA1 activation, we expressed dTRPA1 panneurally and recorded excitatory junctional potentials in muscles in response to warmed (29°C) saline. Finally, we present inexpensive techniques for delivering heat pulses to activate dTRPA1 in the neuronal groups we observed previously while flies are freely behaving. We suggest how to film and quantify resulting behavioral phenotypes with limited resources. Activating all neurons with dTRPA1 caused tetanic paralysis in larvae, while in adults it led to paralysis in males and continuous uncoordinated leg and wing movements in females. Activation of cholinergic neurons produced spasms and writhing in larvae while causing paralysis in adults. When a single class of nociceptive sensory neurons was activated, it caused lateral rolling in larvae, but no discernable effects in adults. Overall, these exercises illustrate principles of modern genetics, neuroanatomy, the ionic basis of neuronal excitability, and quantitative methods in neuroethology. Relatively few research studies have used dTRPA1 to activate neural circuits, so these exercises give students opportunities to test novel hypotheses and make actual contributions to the scientific record.
authors.affiliation: Department of Zoology, University of Cambridge, Cambridge, UK CBQ 3EJ.
authors.forename: Jimena
authors.initials: J
authors.lastname: Berni
authors.forename: Alistair M
authors.initials: AM
authors.lastname: Muldal
authors.forename: Stefan R
authors.initials: SR
authors.lastname: Pulver
keywords: Behavior; Drosophila; GAL4-UAS system; Neuromuscular Junction (NMJ); TRPA1; neurogenetics