Figure 2
Temperature-dependent modulation of cellular activity by Gr28bD. (A) Left panel: dorsal view of adult Drosophila dissected to expose ventral nerve cord motor neurons to saline bath. Right panel: composite image of epifluorescence (green on black) and two-photon 3D reconstruction (gray scale) of abdominal neuromere neurons expressing GCaMP6f. The green trace is an example of single-cell activity, as obtained via epifluorescence from the indicated region of interest. (B) Single-cell fluorescence signals (blue traces) obtained in response to a temperature step (red traces), analyzed by continuous wavelet-transform (spectrogram). Top panel: representative data from control flies expressing only GCaMP6f in motor neurons. Middle and bottom panels: representative data sets illustrating two types of temperature responses in flies expressing GCaMP6f + Gr28bD in motor neurons. (C) Change in average fluorescence between low and high temperature. ΔF/F was calculated as (FH−FL)/FL, where FL and FH are the average fluorescence intensities (in arbitrary units) for a given cell, at low and high temperature. Data points are mean ± SDV (n = 21 cells from five flies for GCaMP6f, and n = 20 cells from six flies for GCaMP6f + Gr28bD). The blue lines represent mean ± SDV for each data set. ΔF/F = −0.1524 ± 0.1358 in control flies and 0.03656 ± 0.10 in GR28bD-expressing flies (unpaired t-test; p-value < 0.00001).
