Tβh gal4

Flies were reared on standard cornmeal-yeast medium under a 12 hr:12 hr dark:light cycle at 25°C and 60% humidity. Flies carrying a dTrpA1 transgene were raised at 21°C. UAS-TNTE and UAS-impTNT were kindly provided by Dr Cahir O’Kane (University of Cambridge). UAS-dTRPA1 was a gift from Dr Paul Garrity (Brandeis University). Dilp2-GAL4 line, trans-Tango line, and elav-GS line were provided by Dr Yi Zhong (Tsinghua University), DskFlp and DskRNAi lines were provided by Dr Yufeng Pan (Southeast University). UAS > stop > myr::GFP (pJFRC41 in attP5) was a gift from Gerald Rubin, UAS > stop > kir^eGFP was provided by Dr Yi Rao, pC1-ss1 and pC1-ss2 were provided by Dr Kaiyu Wang. The following lines were obtained from the Bloomington Drosophila Stock Center: R71G01-GAL4 (BL#39599), R71G01-LexA (BL#54733), TβH-GAL4 (BL#45904), TRIC line (BL#61679), UAS-Kir2.1 (BL#6595 and BL#6596), UAS-mCD8::GFP (BL#5137), UAS > stop > dTrpA^myrc (BL#66871). Lexo-CD4-spGFP11/CyO; UAS-CD4-spGFP1-10/Tb was previously described (Gordon and Scott, 2009 (link)).

Drosophila stocks used in this study were obtained from the following sources: the tyramine-β-hydroxylase (TβH) null mutant line8 (link), Tβh^nM18 (supplied by Henrike Scholz, Cologne, Germany), was described previously8 (link). This line was backcrossed several times with w¹¹¹⁸, the control line for all corresponding experiments. Tdc2^RO54 as well as flies with the matching genetic background were provided by Jay Hirsh (University of Virginia, USA) and have been described previously40 (link). The following transgenic lines were from the Bloomington Drosophila Stock Center: Tβh-GAL4 (#45904), Tdc2-GAL4 (#9313), UAS-dTrpA1 (#26263) and UAS-ChR2 (#9681). Unless otherwise stated, the flies were raised on standard medium at 25 °C with 50–60% relative humidity under a 12:12 h light/dark cycle as described earlier41 (link). All assays utilised virgin females at a density of 20–25 adults per vial.