Double labeling with antiserum to sNPFp and Gad1 -Gal4-driven GFP. Single confocal sections (horizontal views; anterior to top) are shown. A – C. No colocalization of the markers were seen in neurons of the larval ventral nerve cord (A dorsal aspect, B ventral aspect). The dorsal DP neurons in first abdominal ganglion (arrow in A) did not colocalize markers, although similar neurons expressing Gad1-Gal4 were seen in adjacent neuromeres. In C1-2 details of A1 are shown. D1 – 2. The sNPF-IR processes in neuropil overlap Gad1-Gal4 expressing branches in lateral neuropil of abdominal ganglia. Also the posterior neuropil (terminal plexus) in A8/9 (arrow) expresses both markers but, without colocalization (see also D2'). E. In the larval brain no colocalized markers were detected in neuronal cell bodies. The colocalization of Gad1-Gal4 and sNPF in the mushroom body lobes (aL) may be irrelevant here, since the Gad1-Gal4 does not support a GABAergic phenotype (see ). Most Gad1 expressing cell bodies are in the subesophageal ganglion (SEG) in this optical section. F1 – 2. In the adult brain colocalized markers (asterisks) can be seen in a group of dorsal median neurons in the protocerebrum (in the DMN cluster in Fig. 7A). G. In the dorso-lateral protocerebrum at the base of the Kenyon cell cluster (KC) some neurons (arrows) colocalize sNPF and Gad1-Gal4 (in the DLN2 cluster in Fig. 7A).