Figure 3. Electrophysiological effects of acutely activating R23E10 neurons.

(A) Setup for recording in vivo adult Drosophila electrophysiology with whole-cell patch clamp (orange). (B) dFB neuron schematic showing whole-cell recordings targeted to R23E10-Gal4 cell bodies. Injecting current in a stepwise manner causes firing in these neurons (bottom left panel), which are heterogeneous in their endogenous firing patterns (bottom right panel). Scale bars indicate 10 mV and 100 ms. (C) Example traces (left) of a CsChrimson-expressing (UAS-CsChrimson/+;R23E10-Gal4/+ with ATR) spiking cell (top) and non-spiking cell (middle), and of a non-CsChrimson-expressing (R23E10-Gal4/+ with ATR) cell (bottom) when exposed to constant red light (red shading). Superimposed traces of the corresponding types from multiple cell recordings (right panel, top to bottom: n = 10, n = 6, n = 6). Solid red lines indicate mean values. (D) Boxplots show median membrane potentials for CsChrimson-expressing cells (blue) and non-CsChrimson expressing cells (gray, n = 6) before, during and after constant light stimulation (***p<0.001, Friedman test with Dunn’s multiple comparisons to pre-stimulus condition). See also Figure 3—figure supplements 1 and 2.

Figure 3—figure supplement 1. 1Hz stimulation.

(A) Example traces (left) of a CsChrimson-expressing spiking cell (top), a CsChrimson-expressing non-spiking cell (middle), and a non-CsChrimson-expressing cell (bottom) when exposed to 1 Hz light pulse with 5 ms exposure per pulse (orange shading). Superimposed traces from all 120 trials for the corresponding cells shown on the left (right). Solid yellow lines indicate mean values. (B) Boxplots show median membrane potential for CsChrimson-expressing cells (cyan, n = 13) and for non-CsChrimson-expressing cells (gray, n = 4) before, during, and after 1 Hz light stimulation (****p<0.0001, Friedman test with Dunn’s multiple comparisons to pre-stimulus condition).

Figure 3—figure supplement 2. Whole-cell patch physiology in wildtype and INX6 knockdown flies.

Top panels show combined traces (yellow lines indicate mean values) for all 120 pulses for all CsChrimson-expressing R23E10 neurons that showed the presence of evoked spikes (red arrows) in response to the light stimulus pulses at 1 Hz with 5 ms exposure time each pulse, and the absence of any evoked spikes for the wildtype controls (right, 500 ms exposure for R23E10-Gal4 > UAS CsChrimson with no ATR), with spiking events presented as raster plots (middle), and corresponding peristimulus time histograms (bottom).