Fig 6. Co-application of sNPF with other Drosophila peptides suggests that sNPF and PDF may have a unique interaction.
(A) Average inverse FRET traces (CFP/YFP) of IPCs reflecting intracellular cAMP changes. 10−5 M synthetic sNPF was co-applied with PDF, SDNFMRFa, AKH (adipokinetic hormone), DTK (Drosophila tachykinin), Ast-C (Allatostatin-C) /10-5M for each/ or 10−5 M Ast-C was added alone. The black arrow indicates the application point of the different substance. NKH: adenylate cyclase activator, used as positive control. HL3: hemolymph-like saline, used as negative control. (HL3) (B) Maximum inverse FRET changes quantified for each individual neuron and averaged for each treatment from 100 s until 1000 s. Statistical comparison revealed that co-application of sNPF with SDNFMRFa, AKH, and DTK resulted in a significant decrease of cAMP levels compared to the sNPF+PDF co-application. sNPF+Ast-C led to a significant increase in the level of cAMP, however similar change was observed in the case of Ast-C application alone. (C) A close up of the immediate cAMP level changes occurring from the application point until 200 s. (D) Maximum inverse FRET from 100 s until 200 s. The legend indicates the color code of the different treatments and the number of neurons in the dissected brains (in parentheses) considered in this analysis. ***p<0.001, **p<0.01, *p<0.05, n.s. not significant.