Figure 4. MS1 neurons in the SOG are octopaminergic.
(A) Confocal projection of a whole-mounted MS1> CD8::GFP male (left) or female (right) adult central brain. Antibodies against GFP (green) and Bruchpilot (BRP, magenta) were used for immunostaining. Scale bar: 100 µm. (B) Confocal projection of the ventral nerve cord of a male or female expressing CD8::GFP under the control of MS1-Gal4. Scale bar: 50 µm. (C) Top: confocal projection of a male central brain expressing RFP (magenta) in MS1 neurons (using Gal4-UAS) and GFP (green) in Tdc2 neurons (using LexA-LexAOp). Scale bar: 100 µm. Bottom: Magnified view of the SOG region indicated by the rectangle in the corresponding image in the top row. Arrows point to neurons co-expressing MS1>RFP and Tdc2>GFP. Scale bar: 50 µm. (D) Expression of NaChBac::GFP in all MS1 neurons (left, MS1-Gal4, Tdc2-LexA/UAS-NaChBac) and in the non-octopaminergic subset (right, MS1-Gal4, Tdc2-LexA/UAS-NaChBac; LexAop-Gal80). Expression of Gal80, a suppressor of Gal4, in TDC2 neurons removed NaChBac::GFP expression specifically in the SOG. Scale bar: 100 µm. (E) Sleep profile of male flies of the indicated genotypes. N = 75–78. (F) Sleep profile of males in which MS1 neurons were activated with TrpA1 expression at 28°C in an iso31 control (left), Tbhnm18 mutant (middle), or Oamb286 mutant (right) background. N = 20–165. One-way ANOVA followed by Dunnett post hoc test relative to MS1>NaChBac, Tdc2-LexA/+ flies (E) or both parental controls (F).