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. 2020 Nov 23;9:e59502. doi: 10.7554/eLife.59502

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© 2020, Deutsch et al

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Figure 8. — (A) Left: Activation of neurons in the pC1-A line and imaging of neural activity in all Dsx+ cell bodies. Activity in Dsx+ pC1 somas is shown 10 s before stimulus onset and 5 min after stimulus onset for an example experiment. Right: Normalized activity of Dsx+ pC1 somas ((F(t) - F₀)/σ_o; F₀ and F(t) are mean Fluorescence during baseline and fluorescence over time, respectively). Chrimson and TdTomato are expressed in pC1-A cells. GCaMP6s is expressed in Dsx+ cells. pC1 somas imaged following pC1-A activation (along with controls) are both shown if Ft1 > 3σ_o (σ_o - standard deviation during baseline, Ft1 is the mean fluorescence during t1) (see (B) for full set of imaged pC1 neurons). (B) Left: Mean Calcium response in Dsx+ pC1 somas during t1 (x-axis) versus during t2 (y-axis) for pC1-A activation and control. Normalized activity is defined as (F - F_o)/σ_o, where F_o is the mean activity during baseline, σ_o is the standard deviation during baseline, and F is the mean activity during t1 for x and t2 for y. Each dot represents a single segmented soma. Dots above the dashed line represent persistent responses following activation. All imaged pC1 neurons are shown (n = 8 flies, 58 ROIs for pC1-A, n = 5 flies, 25 ROIs for controls). Right: Example traces of (F - F_o)/F_o from two individual pC1 cells, showing different response decays after stimulus offset (matching the results from Figure 7D). Corresponding points are enlarged and marked in green and purple in B (left). (C) Top: Activation of neurons in the pC1-A line and imaging of neural activity in Fru+ cell bodies. Bottom: Spatial pattern of Fru+ cell bodies imaged (here, a left hemibrain is shown (Z-projection of Fru+ neurons expressing GCaMP6s)). Fru+ cell body groups 1 and 2 are defined based on their spatial location - group 1 likely contains Fru+ pC2 and aIPg neuronal cell bodies, whereas group 2 likely contains Fru+ pC1 cell bodies (Figure 8—figure supplement 2). (D) Normalized activity of Fru+ somas from group 1 (Top) or group 2 (Bottom). (E) Mean activity in Fru+ cell bodies from group 1 (left, n = 9 flies, 46 ROIs for pC1-A, n = 5 flies, 13 ROIs for control) and group 2 (right, n = 9 flies, 37 ROIs for pC1-A, n = 5 flies, 24 ROIs for control) during t1 (x-axis) versus during t2 (y-axis) following pC1-A activation and in controls (see Key Resources Table for full genotypes). Data were analyzed and plotted as in (B). Dots above the dashed line represent persistent responses following activation.

Figure 8—figure supplement 1. — (A) Left: Activation of neurons in the pC1-A line and imaging of neural activity in all Dsx+ cell bodies. Activity in Dsx+ pC1 somas is shown 10 s before stimulus onset and 5 min after stimulus onset for an example experiment. Right: Normalized activity of Dsx+ pC1 somas ((F(t) - F₀)/σ_o; F₀ and F(t) are mean Fluorescence during baseline and fluorescence over time, respectively). Chrimson and TdTomato are expressed in pC1-A cells. GCaMP6s is expressed in Dsx+ cells. pC1 somas imaged following pC1-A activation (along with controls) are both shown if Ft1 > 3σ_o (σ_o - standard deviation during baseline, Ft1 is the mean fluorescence during t1) (see (B) for full set of imaged pC1 neurons). (B) Left: Mean Calcium response in Dsx+ pC1 somas during t1 (x-axis) versus during t2 (y-axis) for pC1-A activation and control. Normalized activity is defined as (F - F_o)/σ_o, where F_o is the mean activity during baseline, σ_o is the standard deviation during baseline, and F is the mean activity during t1 for x and t2 for y. Each dot represents a single segmented soma. Dots above the dashed line represent persistent responses following activation. All imaged pC1 neurons are shown (n = 8 flies, 58 ROIs for pC1-A, n = 5 flies, 25 ROIs for controls). Right: Example traces of (F - F_o)/F_o from two individual pC1 cells, showing different response decays after stimulus offset (matching the results from Figure 7D). Corresponding points are enlarged and marked in green and purple in B (left). (C) Top: Activation of neurons in the pC1-A line and imaging of neural activity in Fru+ cell bodies. Bottom: Spatial pattern of Fru+ cell bodies imaged (here, a left hemibrain is shown (Z-projection of Fru+ neurons expressing GCaMP6s)). Fru+ cell body groups 1 and 2 are defined based on their spatial location - group 1 likely contains Fru+ pC2 and aIPg neuronal cell bodies, whereas group 2 likely contains Fru+ pC1 cell bodies (Figure 8—figure supplement 2). (D) Normalized activity of Fru+ somas from group 1 (Top) or group 2 (Bottom). (E) Mean activity in Fru+ cell bodies from group 1 (left, n = 9 flies, 46 ROIs for pC1-A, n = 5 flies, 13 ROIs for control) and group 2 (right, n = 9 flies, 37 ROIs for pC1-A, n = 5 flies, 24 ROIs for control) during t1 (x-axis) versus during t2 (y-axis) following pC1-A activation and in controls (see Key Resources Table for full genotypes). Data were analyzed and plotted as in (B). Dots above the dashed line represent persistent responses following activation.