Figure 2: DAergic PPL2ab neurons regulate male courtship sustainment.
From: PPL2ab neurons restore sexual responses in aged Drosophila males through dopamine
Representative images showing the expression pattern of the Gal4 driver lines: (a) murashka-1-Gal4, (b) NP3024-Gal4, (c) NP5945-Gal4 and (d) LG121-LexA; driver expression patterns revealed by UAS-mCD8::GFP (or LexAop-mCD8::GFP) in the adult brain (10-day-old) are shown (green in a–d shows the PPL2ab cell bodies indicated by the arrows and POT track indicated by the arrowheads; the inset box shows the expression pattern in a single calyx). The neuropil was immunostained with an anti-DLG antibody (magenta in a–d). The cell bodies of PPL2ab neurons (green in a1-d1) were labelled with four independent drivers; PPM1/2 neurons (green in a2-c2) were labelled by murashka-1-Gal4, NP3024-Gal4 and NP5945-Gal4; and PPM3 neurons (green in b3) were labelled by the NP3024-Gal4; all of these cell were DAergic as indicated by TH-antibody immunostaining (magenta in a1-2, b1-3, c1-2 and d1). Scale bars, 20 μm. (e,f) The results for courtship index and courtship bout length towards 3-day-old intact CS females or decapitated CS females are shown. ANOVAs were used to compare genetically manipulated 10-day-old male flies (murashka-1-Gal4>UAS-TH, NP3024-Gal4>UAS-TH, NP5945-Gal4>UAS-TH, and LG121-LexA>LexAop-TH) with the corresponding driver and effector heterozygous controls. Significant differences in courtship index (e) and courtship bout length (f) were observed. Each column represents the mean of 18 tests. The error bars indicate+s.e.m. The means for columns followed by the same letters were not significantly different according to the P<0.05 threshold by a one-way ANOVA followed by Tukey’s test per grouped columns (separated by a dashed line). (g,h) Two types of PPL2ab neurons from murashka-1-Gal4 flip-out clones (green) and neuropil were immunostained with the anti-DLG antibody (magenta). Genotypes in (a–f): (1) +/y;+/+;murashka-1-Gal4/UAS-TH; (2) +/y;+/+;murashka-1-Gal4/+; (3) +/y;+/+;NP3024-Gal4/UAS-TH; (4) +/y;+/+;NP3024-Gal4/+; (5) +/y;+/+;NP5945-Gal4/UAS-TH; (6) +/y;+/+;NP5945-Gal4/+; (7) +/y;+/+;+/UAS-TH; (8) +/y;LexAop-TH/+;murashka-1-Gal4/+; and (9) +/y;LexAop-TH/+;+/+. Genotypes in (g,h): hs-FLP/y; +/+; UAS>rCD2,y+>mCD8::GFP/murashka-1-Gal4.
