A Genetic Toolkit for Dissecting Dopamine Circuit Function in Drosophila

Abstract

SUMMARY The neuromodulator dopamine (DA) plays a key role in motor control, motivated behaviors, and higher-order cognitive processes. Dissecting how these DA neural networks tune the activity of local neural circuits to regulate behavior requires tools for manipulating small groups of DA neurons. To address this need, we assembled a genetic toolkit that allows for an exquisite level of control over the DA neural network in Drosophila. To further refine targeting of specific DA neurons, we also created reagents that allow for the conversion of any existing GAL4 line into Split GAL4 or GAL80 lines. We demonstrated how this toolkit can be used with recently developed computational methods to rapidly generate additional reagents for manipulating small subsets or individual DA neurons. Finally, we used the toolkit to reveal a dynamic interaction between a small subset of DA neurons and rearing conditions in a social space behavioral assay.

Social behavior and aging: A fly model

“…188,200 Similarly, both virgin male and virgin females are less social than their mated counterparts. The condition and previous experiences of the fly can affect social spacing.…”

“…Socially isolated flies are more distal, and therefore less social, an effect that is more pronounced in females than in males. 188,200 Similarly, both virgin male and virgin females are less social than their mated counterparts. 188 Recently, our lab found that as D. melanogaster age, social spacing changes in an atypical pattern 118 ( Figure 1D).…”

“…Specifically, tyrosine hydroxylase is important for the response to social isolation in dopaminergic neurons in the PPM2 cluster, in males. 200 Social space can also be affected by proteins located on the post-synaptic membrane. Some of the genes encoding these post-synaptic proteins have also been proposed as candidate genes for neuropsychiatric disorders.…”

Trans-synaptic Fish-lips Signaling Prevents Misconnections between Non-synaptic Partner Olfactory Neurons

“…C15-p65 AD is generated by co-injecting a gRNA (cloned in pU6-BbsI-chiRNA) targeting the end of the last exon and donor sequence containing homology arms, p65(AD)::Zip+, and 3XP3-RFP-SV40 (32). Mz19-Gal4 DBD is generated using the HACK strategy (36). We replaced QF2 sequence of pBPGUw-HACK-G4>QF2 with the DNA binding domain of GAL4 cloned from pBS-KS-attB2-SA(0)-T2A-Gal4DBD-Hsp70 polyA and injected it to Mz19-GAL4; vas-Cas9 embryos.…”

“…To achieve specific GAL4 expression, we utilized the split-GAL4 strategy (32,33) to generate new driver lines specific to VA1d and DC3 PNs. We inserted the hemidriver p65 AD component after the last coding exons of C15, a transcription factor that is only expressed by PNs derived from the anterodorsal neuroblast (adPNs) (34), and converted Mz19-GAL4 to Mz19-GAL4 DBD using the HACK strategy (35,36). When we intersected these two components, expression of functional GAL4 was restricted to Mz19+ adPNs (VA1d and DC3) ( Fig.…”