Figure 1. ISNs relay information to the Pars Intercerebralis.

(A) Temporal consumption assay screen for water ingestion using. RNAi targeting different neurotransmitter pathways. UAS-RNAi + or − ISN-Gal4. RNAi against: nSynaptobrevin (nSyb), tryptophan hydroxylase (TRH), choline acetyltransferase (ChAT), tyrosine beta-hydroxylase (TBH), histamine decarboxylase (HDC), vesicular monoamine transporter (VMAT), glutamic acid decarboxylase 1 (GAD1), dopa decarboxylase (DDC), Drosophila vesicular glutamate transporter (DVGlut), short neuropeptide F (sNPF), vesicular GABA transporter (VGAT), Tyrosine decarboxylase 2 (TDC2), Drosophila insulin like peptide 1 (dILP1), Drosophila insulin like peptide 2 (dILP2), Drosophila insulin like peptide 3 (dILP3), Drosophila insulin like peptide 4 (dILP4), Drosophila insulin like peptide 5 (dILP5), Drosophila insulin like peptide 6 (dILP6), Drosophila insulin like peptide 7 (dILP7). Represented are the mean, and the 10–90 percentile; data was analyzed using Kruskal-Wallis test, followed by multiple comparisons against the RNAi control; p-values were adjusted using False Discovery Rate. n=8–39 animals/genotype except nSyb positive control (70–72). (B) Temporal consumption assay for 1M sucrose or water using RNAi targeting dILP3 or amontillado in ISNs. Sucrose assay: Kruskal-Wallis test followed by Dunn’s multiple comparison tests against ISN control and respective RNAi control. Water assay: ANOVA, Šídák’s multiple comparison test to ISN control and respective RNAi control. n=48–52 animals/genotype. (C) ISNs reconstruction from FAFB volume. (D) Light microscopy image of ISN-Gal4 registered to JFRC2010. (E) ISN postsynaptic neurons based on synapse predictions using FAFB volume (Zheng et al. 2018) and connectome annotation versioning engine (CAVE, Buhmann et al. 2021, Heinrich et al. 2018). Left: 10 postsynaptic neurons, right: postsynaptic neurons BiT, Cowboy, Handshake and DSOG1. *p<0.05, ***p<0.001