FMRFa receptor stimulated Ca2+ signals alter the activity of flight modulating central dopaminergic neurons in Drosophila melanogaster
Fig 6
Expression of FMRFaRRNAi in dopaminergic neurons reduces membrane depolarization.
(A) Mean trace (±SEM) of negative change in fluorescence (-ΔF/F) observed upon KCl stimulation of THD1 neurons expressing a fluorescent-based membrane voltage indicator, Arclight in the indicated genotypes, THD1;TubGAL80ts>Arclight, Control in black; THD1;TubGAL80ts>Arclight;FMRFaRRNAi, in red. (B) Area under the curve and (C) Peak (-ΔF/F) quantified from (A) were significantly different in the FMRFaR knockdown neurons as compared to their genotypic controls. Numbers below each box plot indicate total number of cells imaged from a minimum of 5 independent brains (One-way ANOVA followed by post-hoc Tukey’s test; the same alphabet above each bar represents statistically indistinguishable groups; different alphabet represents p<0.05). (D) Representative images of neuronal depolarization as observed by a decrease in Arclight fluorescence in the same genotypes shown in (A). With FMRFaR knockdown (2nd row), few cells responded normally (yellow arrow), whereas most others showed no response or a minimal response to the depolarization stimulus (pink arrow). Scale bars represent 50 μm. (E) Rescue of flight bout durations by expression of dTrpA1 in the genetic background of FMRFaRRNAi in 6 and 8 day old adult THD1 marked neurons (THD1;TubGAL80ts>dTrpA1;FMRFaRRNAi compared to THD1;TubGAL80ts>FMRFaRRNAi; n≥30, **p<0.01, Mann-Whitney U-test).