Skip to main content
. 2020 Nov 6;9:e62297. doi: 10.7554/eLife.62297

Figure 5. Carbachol-stimulated Dopamine release at the axonal terminals of PPL1-γ2α′1 neurons is attenuated by expression of IP3RDN.

(A) Representative images of a presynaptic marker synaptotagmin GFP (syt.eGFP) in lobes of the Mushroom body (top). Quantification of syt.eGFP fluorescence normalized to mRFP is shown below in the indicated MB regions marked by THD’GAL4. N = 6 brains, n.s. not significant at p < 0.05 by two tailed Mann-Whitney U test. Scale bars indicate 50 μm (B) Carbachol stimulated dopamine release at PPL1- γ2α′1 termini visualized by changes in GRABDA fluorescence in a representative mushroom body lobe from brains of the indicated genotypes. Images were acquired at 2s per frame. Brighter fluorescence denotes increase in Dopamine. Scale bars = 10 μm (C) Average traces (± SEM) of normalized GRABDA fluorescence responses (ΔF/F) from the right lobe of each brain upon addition of Carbachol (Green line) (panel on the left). Images were acquired at 2s per frame; Quantification of area under the curve was from the point of stimulation at 160s and up to 250 s. Box plots and symbols are as described in Figure 2A. MB296B GAL4 > UAS GRABDA N = 10 cells from 10 brains; MB296B GAL4 > UAS ItprDN; UAS GRABDA N = 8 cells from 8 brains. **p< 0.05, Mann-Whitney U test.

Figure 5—source data 1. GRABDA fluorescence (ΔF/F) traces from axonal termini of MB296BGAL4 marked DANs in individual brains (Black -MB296B GAL4>UAS GRABDA; Red-MB296B GAL4>UAS ItprDN; UAS GRABDA).

Figure 5.

Figure 5—figure supplement 1. Carbachol evoked Ca2+ responses in the soma of PPL1- γ2α′1 DANs marked by MB296BGAL4.

Figure 5—figure supplement 1.

(A) Carbachol stimulated change in GCaMP6m fluorescence (ΔF/F) in PPL1- γ2α′1 cells. Representative images of GCaMP6m fluorescence at the indicated time points (top panel). Warmer colors denote increase in [Ca2+]. Average response (± SEM) of normalized changes in GCaMP6m fluorescence (ΔF/F) in MB296BGAL4 marked PPL1 cells upon addition of Carbachol from N=7 cells from 7 brains. Images were acquired at 2s per frame. (B) Average traces (± SEM) of normalized changes in GRABDA fluorescence (ΔF/F) from axonal termini of MB296BGAL4 marked DANs that responded below the arbitrary threshold (ΔF/F=0.05) upon addition of Carbachol (Green line). Images were acquired at 2s per frame (Top panel). Quantification of area under the curve (AUC) from the point of stimulation 160s up to 250s. Boxplots and symbols are as described in Figure 2A. MB296B GAL4 > UAS GRABDA N = 3 brains; MB296B GAL4 > UAS ItprDN; UAS GRABDA N = 9 brains, *p< 0.05, Mann-Whitney U test (lower panel).