Skip to main content
. 2025 Sep 3;19:1683967. doi: 10.3389/fnins.2025.1683967

Figure 1.

Diagram detailing experimental data. Panel A shows a genetic scheme using the GAL4/UAS system with luciferase expression. Panel B presents two line graphs (G0338 and G0431) showing luminescence over nine days with orange and purple data points, respectively. Panel C includes dot plots showing periodicity in hours over nine days for G0338 and G0431 with box plots indicating 20, 25, 30, and 35 hours. Panel D shows a bar graph comparing performance index values for iso1, time, iso1, and cyco, with bars in black, gray, and blue.

The LABL reporter oscillates in DAL neurons and circadian mutants have normal 3-day memory. (A) The LABL reporter system. Cartoon of three transgenes (shown as line cartoons) that are present in the same fly. The first transgene expresses the transcription factor Gal4 in a DAL-specific anatomical pattern. The Gal4 protein acts on UAS sequences to make the FLP recombinase (second line cartoon). The third transgene contains a fragment from the period promoter (perP) and a FLP-out cassette (triangles flanking mCherry, followed by stop codons in all reading frames) upstream of the coding region for luciferase (missing its ATG start codon). The black arrow signifies the start of transcription and translation. In the presence of FLP, recombination at the FRT sites results in deletion of one FRT site and the intervening sequences, including the translation termination codons (fourth line cartoon). (B) FLP expression in DAL neurons results in oscillating reporter activity. When the LABL system is driven using the DAL-specific Gal4 lines (G0338 [left plot in orange] or G0431 [right plot in magenta]), transcriptional activity oscillates. Luminescence (in arbitrary units) is plotted as a function of time. Daytime and nighttime periods are shown in light and dark gray and the vertical black lines indicate a 24-h period. The best fit maxima and minima for each 24-h period are shown with circles. (C) LABL oscillates with an approximate 24-h periodicity in DAL neurons. Data from the LABL experiments (n = 4) is analyzed as before (Johnstone et al., 2022). Morlet wave analysis for the entire 9 days shows that the periodicity of both reporters is about 24 h. Confidence intervals are shown in shades of color and indicated above the figure. The violin plots indicate the spread of all the data points. (D) Mutations in tim and cyc do not significantly affect 3d memory. The 3d performance indices for wild-type (iso31) and mutant stocks are plotted as a function of genotype. There is no significant difference in either mutant when compared to its wild type control.