Fig. 3: Glutamatergic DN1as primarily send inhibitory projection to E cells.
a DN1as are glutamatergic neurons. Anti-RFP (glutamate) and anti-GFP (DN1a) staining were visualized in the brain of flies. Scale bars, 20 μm. b Schematic of the major downstream neurons of DN1as reconstructed in fly brain template. The left panel is the EM reconstruction of DN1as and downstream neurons. The right panel displays a Sankey histogram depicting the synaptic connections between DN1as and major postsynaptic neurons, as determined by the Hemibrain connectome dataset. a: anterior. l: latter. v- ventrolateral. c Identifying postsynaptic connections of DN1as using trans-Tango. From left to right: Presynaptic DN1as (yellow), merged images, and postsynaptic trans-Tango signal (cyan / black). The genotype was labeled above the panel. Scale bars, 20 μm. d Representative PER and GFP double staining of DN1a-spl > trans-Tango brain. Anti-PER (cyan) and anti-GFP (LNd: yellow) staining were visualized in the brain of flies. White circle indicates LNds. Scale bar, 50 μm. e Double staining of GFP (cyan) and RFP (yellow) in Dvpdf-LexA > LexAop-GFP, DN1a-spl>UAS-tdTomato brain. Scale bars, 50 μm. f Representative pseudocolored images of calcium responses of LNds before and after ATP perfusion activation of DN1as (left); Scale bars, 10 μm. Representative GCaMP traces (ΔF/F ± SEM) of LNds in response to DN1as activation (right, n = 12). Con: grey; Exp: green. g Quantification of relative max calcium activities of LNds in (f). Data are presented as mean in MaxΔF/F0 ± SEM in the histogram; 2-tailed t-test; ****p < 0.0001, p = 0.000145. n = 12.
