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. 2017 Mar 23;6:e24992. doi: 10.7554/eLife.24992

Figure 4. Silencing and activating the IR60b neuron alters feeding time.

(A) Initial analysis of total feeding time of IR60b-GAL4/UAS-TNT, IR60b-GAL4/+, and UAS-TNT/+ flies when 300 mM sucrose was delivered in the pharyngeal pumping assay (**p<0.01, one-way ANOVA, Bonferroni post-test, n = 16–24). (B) Total feeding time of silenced flies at a range of sucrose concentrations (*p<0.05, ***p<0.001: two-way ANOVA, Bonferroni post-test, n = 18–30). (C) Pumping rates of silenced flies at different sucrose concentrations. No differences were observed in pumping rates among different genotypes or doses (two-way ANOVA, Bonferroni post-test, n = 16–27). (D) Total feeding time of silenced flies when tested with glucose, fructose, or trehalose. No increases in sugar consumption were observed in IR60b-GAL4/UAS-TNT flies when compared to controls (one-way ANOVA, Bonferroni post-test, n = 25–38). (E) UAS-Chrimson (CHR) activation: total feeding time with 300 mM trehalose in IR60b-GAL4/UAS-Chrimson, IR60b-GAL4/+, and UAS-Chrimson/+ flies under low-light conditions, either with or without application of red light. IR60b-GAL4/UAS-Chrimson flies show a decrease in feeding when IR60b neurons are activated with red light (*p<0.05, two-way ANOVA, Bonferroni post-test, n = 12–24). Under red light, feeding time for IR60b-GAL4/UAS-Chrimson flies is lower than for IR60b-GAL4/+ and UAS-Chrimson/+ controls (*p<0.05, two-way ANOVA, Bonferroni post-test, n = 12–24).

DOI: http://dx.doi.org/10.7554/eLife.24992.008

Figure 4.

Figure 4—figure supplement 1. Expression of UAS-TNT driven by IR94f-GAL4 also causes overconsumption.

Figure 4—figure supplement 1.

(A) Total feeding time of 300 mM sucrose for IR94f-GAL4/UAS-TNT, IR94f-GAL4/+, and UAS-TNT/+ flies in the pharyngeal pumping assay. IR94f-GAL4/UAS-TNT females exhibited increased total feeding time compared to heterozygote controls (*p<0.05, ***p<0.001, one-way ANOVA, Bonferroni post-test, n = 12–16). These results support the conclusion from mapping data that IR60b and IR94f co-express in the same pharyngeal neuron. The IR94f-GAL4 and UAS-TNT constructs were each backcrossed for at least five generations against our control w Canton S genetic background before the experiment.
DOI: http://dx.doi.org/10.7554/eLife.24992.009
Figure 4—figure supplement 2. Optogenetic activation of IR60b neurons does not reduce feeding on sucrose.

Figure 4—figure supplement 2.

The feeding time of IR60b-GAL4/UAS-Chrimson flies in red light is not lower than that of parental controls during delivery of 300 mM sucrose (one-way ANOVA, n = 36–49), suggesting that activation of IR60b neurons by sucrose is sufficiently strong that red light does not produce a demonstrable incremental effect.
DOI: http://dx.doi.org/10.7554/eLife.24992.010