Validation of an Optogenetic Approach to the Study of Olfactory Behavior in the T-Maze of Drosophila melanogaster Adults

Abstract

Simple SummaryThe fruit fly (Drosophila melanogaster) has been used as a model organism to study the olfactory system of insects thanks to the wide range of genetic tools available in this species. Among these tools, optogenetics allows the immediate alteration of the functioning of certain cells with light by the targeted expression of light receptor proteins in these cells. Thus, by successively expressing these receptors in different elements of the behavioral circuit, it is possible to evaluate their effect on the final behavior of the organism. However, the use of optogenetics to dissect the receptor elements of adult olfactory behavior presents a challenge because most odorants elicit gradual attraction or avoidance depending on their concentration, complicating the representative substitution of odor by light. In this work, we explore a dual excitation model in which the subject responds to various odorant concentrations while the olfactory receptor neurons are activated by light. The dose–response curve in these flies remains odorant concentration dependent, but with reduced sensitivity compared to olfactory stimulation alone. The existence of an effect associated with each of the two stimuli, odor and light, allows us to explore the quantitative contribution of the receptor elements to olfactory behavior also by optogenetics.Optogenetics enables the alteration of neural activity using genetically targeted expression of light activated proteins for studying behavioral circuits in several species including Drosophila. The main idea behind this approach is to replace the native behavioral stimulus by the light-induced electrical activation of different points of the circuit. Therefore, its effects on subsequent steps of the circuit or on the final behavior can be analyzed. However, the use of optogenetics to dissect the receptor elements of the adult olfactory behavior presents a challenge due to one additional factor: Most odorants elicit attraction or avoidance depending on their concentration; this complicates the representative replacement of odor activation of olfactory sensory neurons (OSNs) by light. Here, we explore a dual excitation model where the subject is responding to odors while the OSNs are optogenetically activated. Thereby, we can assess if and how the olfactory behavior is modified. We measure the effects of light excitation on the response to several odorant concentrations. The dose-response curve of these flies still depends on odor concentration but with reduced sensitivity compared to olfactory stimulation alone. These results are consistent with behavioral tests performed with a background odor and suggest an additive effect of light and odor excitation on OSNs.