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. 2015 Jun 17;4:e06914. doi: 10.7554/eLife.06914

Figure 5. Tachykinin-expressing cells in the SEZ are a second order circuit for Gr68a neurons.

(A) Ablation of TK-expressing circuits using two independent Gal4 drivers (TK2 and TK3) removed sensitivity to CH503. Homozygous or trans-heterozygous ΔTK deletion mutants also exhibit a loss of sensitivity to CH503. Rescuing TK expression in two different mutant backgrounds restored the behavioral response to CH503. See Figure 5—figure supplement 2 for parental controls. N = 15–31, Fisher's exact probability test, ns: not significant, ****p < 0.0001. (B, C, E) TK-expressing cells are closely apposed to Gr68a-Gal4 synaptic terminals labeled with synaptobrevin-GFP (syb-GFP) in the SEZ (C) and AMMC (E). No co-localization is observed (Pearson's coefficient: 0). Image represents a maximum intensity Z-series projection. (D, D′) Positive GRASP-GFP signal in the SEZ indicates synaptic connectivity between Gr68a neurons and TK processes. The GFP signal is overlaid on a phase-contrast image of the tissue (arrows). (F, F′) Positive GRASP-GFP signal in the AMMC indicates synaptic connectivity between Gr68a neurons and TK processes. The GFP signal is overlaid on a phase-contrast image of the tissue (rectangle). Scale bar B: 50 μm; all other scale bars: 20 μm.

DOI: http://dx.doi.org/10.7554/eLife.06914.025

Figure 5.

Figure 5—figure supplement 1. Tachykinin is essential for CH503 detection.

Figure 5—figure supplement 1.

Three different TK-Gal4 drivers were tested for their contribution to CH503 processing. Cell ablation or inactivation of neural activity using TK2-Gal4 and TK3-Gal4 (purple vs white) but not TK1-Gal4 (black vs white) removed sensitivity to (S, Z, Z)-CH503. N = 28–36, Fisher's exact probability test, ns: not significant; **p < 0.01; ***p < 0.001.
DOI: http://dx.doi.org/10.7554/eLife.06914.026
Figure 5—figure supplement 2. Parental control lines for tachykinin mutant rescue experiments.

Figure 5—figure supplement 2.

The presence of the Gal4 or UAS transgene is not sufficient to rescue the response to CH503. N = 23–30, Fisher's exact probability test, all results are not significant.
DOI: http://dx.doi.org/10.7554/eLife.06914.027
Figure 5—figure supplement 3. Non-specific diffuse staining is observed in tissue from GRASP negative controls lacking the Gr68a-Gal4 driver.

Figure 5—figure supplement 3.

Left: the brightness of the confocal image is exaggerated in order to visualize the tissue. Right: the fluorescent image is overlaid on a phase-contrast image of the tissue. Scale bar: 50 μm.
DOI: http://dx.doi.org/10.7554/eLife.06914.028