Extended Data Fig. 7: Copulation attempts drive dopaminergic inhibition of the visual threat pathway.
A, ∆F/F0% of LC16 > GRABDA signal of males paired with a male fly. Right: mean ∆F/F0% during 1st min (0-20 s) and at 4 min (220-240 s) (n = 5). B, ∆F/F0% of LC16 > GCaMP6f signal of males pre and post application of 500 µM dopamine. Right: mean ∆F/F0% pre and post time windows (n = 10). C, ∆F/F0% of LC16 > GCaMP7b signal after focal injection of dopamine (blue), saline (grey), or dopamine in LC16 > GCaMP7b; Dop2R-RNAi males (magenta). Scale bar: 5 µm. Right: mean ∆F/F0% pre and post-injection. Saline (n = 5, 3 flies) vs. Dopamine (n = 6, 4 flies) vs. Dop2R-RNAi (n = 5, 3 flies). D, TH genomic region showing sequences upstream of Gal4 for lines C1 and C’. Exons are black boxes. ATG, stop, and i1-i6 mark translational start, stop, and intron sites. “+”, “+/−”, and “−” in the table indicate expression in most, a subset, or none of the neurons in the clusters, respectively. See69. Panel D is adapted from ref. 69, Elsevier. E1-3, ∆F/F0% of LC16>LexAop-GCaMP6f; TH-C1>CsChrimson signal pre and post: E1 threat, E2 threat with LED stimulation, or E3 threat and LED stimulation in control flies only expressing UAS-GCaMP6f in LC16 neurons. Right: Mean ∆F/F0% pre and post time windows (n = 8,8,7). F, Left: LC16 Split-GAL4 driving UAS-spGFP1-10 without (left) or with (right) tagging endogenous Dop2R::GFP11. Right: quantified GFP fluorescence in LC16 cell bodies (left) or axon terminals (right) with or without Dop2R::GFP11. anti-brp (nC82; magenta). (n = 8,10,7,11 brains). GFP fluorescence was normalised to the average fluorescence in the LC16 axon terminals in LC16 > GFP1-10,Dop2R::GFP11 flies. Scale bar: 25 μm. G1-3, ∆F/F0% TRHPMPD > GCaMP6s pre and post threat (G1), threat with 500 µM dopamine (G2), and threat with 500 µM dopamine with additional knockdown of Dop2R in LC16 neurons (G3). Right: mean ∆F/F0% of pre and post-time windows (n = 5,5,7). See legend to Extended Data Fig. 5 for details on graphics and statistics.
