Drosophila Free-Running Rhythms Require Intercellular Communication (original) (raw)
Figure 3
Molecular Oscillations of tim RNA Damp in DD in the Pdf01 Mutant
tim RNA oscillations were examined in the Pdf01 mutant under both LD (A) and different days in DD ([B] and [C]), by whole-mount in situ hybridization. (A), (B), and (C) are representative images from replicas of three experiments.
(A) The left panel is from ZT3, and the right panel is from ZT15. A normal tim oscillation profile is observed compared to that of wild-type (see Figure 2A).
(B) Brains from the Pdf01 mutant in the first day of DD. Left panel, CT3; right panel, CT15. Oscillations are comparable to those in LD.
(C) Brains taken in the fourth day of DD. Six timepoints were taken throughout the circadian day. The sequence of panels from left to right is CT2, 6, 10, 14, 18, and 20, respectively. Wild-type brains (top row) were assayed in parallel with those from the Pdf01 mutant (bottom row). See text for details.
(D) Quantification of (C). Relative intensities are taken from normalized mean pixel intensities. Different clock neuronal groups were quantified independently and compared between wild-type (blue curves) and Pdf01 mutant (purple curves). The panels from left to right are quantification of tim RNA oscillation in the DNs, in the LNds, and in the LNvs. Reduced cycling amplitude and a significant advanced phase were observed in the fourth day of DD. See text for details.