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During olfactory appetitive learning, animals associate an odor, or conditioned stimulus (CS), with an unconditioned stimulus (US), often a sugar reward. This association induces feeding behavior, a conditioned response (CR), upon subsequent exposure to the CS. In this study, we developed a model of this behavior in isolated  Drosophila  brains. Artificial activation of neurons expressing the  Gr5a  sugar-responsive gustatory receptor ( Gr5a  GRNs) induces feeding behavior in starved flies. Consistent with this, we find that in dissected brains, activation of  Gr5a  GRNs induces Ca 2+  transients in motor neurons, MN11 + 12, required for ingestion. Significantly, activation of  Gr5a  GRNs can substitute for presentation of sugar rewards during olfactory appetitive learning. Similarly, in dissected brains, coincident stimulation of  Gr5a  GRNs and the antennal lobe (AL), which processes olfactory information, results in increased Ca 2+  influx into MN11 + 12 cells upon subsequent AL stimulation. Importantly, olfactory appetitive associations are not formed in satiated flies. Likewise, AL-evoked Ca 2+  transients in MN11 + 12 are not produced in  ex vivo  brains from satiated flies. Our results suggest that a starved/satiated state is maintained in dissected brains, and that this  ex vivo  system will be useful for identification of neural networks involved in olfactory appetitive learning.

A Drosophila ex vivo model of olfactory appetitive learning