23e10 gal4

Flies were reared at 25°C and 60% humidity and were kept in a 12-hr light-12-hr dark cycle (or in constant darkness when specified). For thermogenetic activation of neurons, flies were raised and loaded into the visual-recording system at 22°C. Temperature was changed to 28°C when neural activation to place. Flies were backcrossed into a Canton S for at least five generations. UAS-Hid, Dilp2-Gal4, Pdf-Gal4 and MB247-Gal4 were generously provided by Aike Guo (Institute of Biophysics, CAS). Tub > Gal80 > was provided by Jing Wang (UCSD). UAS-StingerGFP and LexAop-tdTomato were gifts from Barry Dickson (Janelia Research Campus, HHMI). UAS-TrpA1 was provided by Paul Garrity (Brandeis University). UAS-mCD8GFP, UAS-StingerGFP, UAS-DscamGFP, UAS-sytGFP, UAS-DenMark, UAS-NachBach, UAS-FRT-stop-FRT-mCD8GFP, 23E10-Gal4, 23E10-LexA, 69F08-Gal4, 18H11-Gal4, LexAop-Gal80, TH-Gal4 and LexAop-Flp were obtained from the Bloomington Stock Center. UAS-5HT2b-RNAi (KK102356) was obtained from the Vienna Drosophila Resource Center (VDRC). Table 1 contains the complete genotypes used in the figures.

All flies were maintained in standard cornmeal–yeast–agar medium at 25 °C. w[1118] (BL5905; wild-type control), Sh[mns] (BL24149), Sh[TKO] (BL76968), sss[P1] (BL16588), sss[MIC] (BL34309), Gad1[L352F] (BL6295), VGAT[MB01219] (BL23022), TrpA1[1] (BL26504), UAS-Sh RNAi #1 (BL53347), UAS-Rdl RNAi (BL52903), ELAV-Gal4 (BL458), ELAV-Gal4; UAS-Cas9 (BL67073), VGAT-Gal4 (II) (BL58980), VGAT-Gal4 (III) (BL58409), 23E10-Gal4 (BL49032), TH-Gal4 (BL8848), TDC2-Gal4 (BL9313), ChAT-Gal4 (BL6798), 121y-Gal4 (BL30815), 30y-Gal4 (BL30818), GAD1-LexA (BL60324), UAS-mLexA-VP16-NFAT (BL66542; CaLexA), UAS-DenMark, UAS-syt.eGFP (BL33065), lexAop-nSyb-spGFP1-10, UAS-CD4-spGFP11 (BL64315; syb:GRASP), UAS-SuperClomeleon (BL59847), and UAS-Epac1-camps (BL25407) were obtained from Bloomington Drosophila Stock Center. Hk[1] (101-119) was obtained from Kyoto Stock Center. UAS-Sh RNAi #2 (v104474) was obtained from Vienna Drosophila Resource Center. GAD1-Gal80, TH-LexA, and UAS-sytGCaMP6s; UAS-P2X₂ were gifts from Y. Li (Chinese Academy of Sciences), H. Tanimoto (Tohoku University), and R. Allada (Northwestern University), respectively.

The UAS-dbt^RNAi (VDRC: v9241) line was from the Vienna Drosophila RNAi center; the dbt mutants, including dbt^S, dbt^L and the UAS-dbt^K/R line were previously described [8 (link),11 (link),31 (link)]; the UAS-dicer (I); tim-GAL4 line was previously described [32 (link)]; the UAS-dicer (II) (BDSC: 24650), dilp2-GAL4 (BDSC: 37516), 23E10-GAL4 (BDSC: 49032), EB-GAL4(BDSC: 44409), Cre;D/Sb (BDSC: 851) and tub-GAL80ts (BDSC: 7108) were from Bloomington Stock Center; the tim-GAL4 (BDSC: 7126) and per-GAL4 (BDSC: 7127) drivers were previously reported [22 (link),33 (link)]; the repo-GAL4 was provided by Dr. Yi Rao (Peking University), the C309 [24 (link),34 ,35 (link)] and 201Y [24 (link),36 (link)] lines were provided by Dr. Yan Li (Institute of Biophysics, Chinese Academy of Science); UAS-CD8: mGFP (BDSC: 5137) [37 (link)], C205 [38 (link)], 44409 [39 (link)], C232 and C819 [40 (link)] were provided by Dr. Li Liu (Institute of Biophysics, Chinese Academy of Science). The mutants dbt^S and dbt^L have been backcrossed with Canton-S for 6 generations and all experiments have been repeated at least 3 times in the manuscript.