FIGURE 6.

FLP³³⁵ and tsh-GAL80 restrictive labeling of the dopaminergic GAL4 line. (A) FLP³³⁵ tsh-GAL80 TH-GAL4 UAS>stop>mCD8::GFP showing exclusive targeting of neurons that are double positive for GFP and 5HT at the abdominal ganglion. (B,C) Two different depths are shown to illustrate the co-localized signals. Tissues were stained with anti-mCD8 (green) and anti-5HT (magenta). Scale Bar = 50 μm. (D–F) Effect of silencing FLP³³⁵ and tsh-GAL80 restrictive labeling of TH-GAL4. (D) Percentage of successful matings in 1 h (FLP³³⁵, TH>TNTin, tsh-GAL80: n = 31; FLP³³⁵, TH>TNT, tsh-GAL80: n = 36). NS, not significant by Fisher’s exact test. (E) Copulation latency (central line indicates the median; FLP³³⁵, TH>TNTin, tsh-GAL80: n = 30; FLP³³⁵, TH>TNT, tsh-GAL80: n = 36). NS, not significant by Mann–Whitney test. (F) Copulation duration (central line indicates the median; FLP³³⁵, TH>TNTin, tsh-GAL80: n = 30; FLP³³⁵, TH>TNT, tsh-GAL80: n = 36). ^∗p < 0.0172 by Mann–Whitney test. (G) Double staining experiment of the FLP³³⁵ tsh-GAL80 restricted GFP expression of TH-Gal4 male reproductive tissues with anti-GFP (green) and anti-5HT (magenta). (H–J) Co-localization of GFP and 5HT signals at the seminal vesicle (H), the accessory glands (I), and the cell-like structure near the testicular duct at the junction between the testes and the seminal vesicle (J). AC, accessory gland; ED, ejaculation duct; SV, seminal vesicle; TD, testicular duct; Te, testes; MeT, median trunk nerve; VNC, ventral nerve cord; Abg, abdominal ganglion.