Figure 3
Temporal alignments of time-series data help to uncover phenotypic abnormalities caused by genetic perturbations. This example compares a control genotype (left panels) and a genetic perturbation caused by the overexpression of EAST(1-1902)-GFP (right panels) that interferes with muscle remodeling. The panels show maximum intensity projections (MIPs) of 3D stacks of dorsal abdominal regions. The control pupa expresses UAS-Grasp65-GFP (green) and UAS-Histone-mKO (red) driven by Mef2-Gal4 to label cytoplasm and nuclei, respectively. The UAS-east(1-1902)-GFP mutant pupa (green nuclei) expresses MHC-tau-GFP (green) to label cytoplasm and UAS-histone-mKO. Time stamp 0:00 (hours:minutes) refers to the onset of PPT, negative time stamps indicate prepupal and positive ones pupal stages. (a, a') At -6:00, no discernible differences are observed. The arrows point in the anterior to posterior direction along the bilateral symmetry axis. Arrow head indicate external oblique muscles that undergo apoptosis during metamorphosis. (b, b') At -2:00, deformation of the external muscle (arrow head) is more pronounced in the control than the mutant animal. (c) Muscle contractions at the onset of PPT lead to blurring of the MIPs. (d) At +12:00, disintegrating external muscles give rise to vesicles (arrow head) while the mutant specimen (d') shows intact external muscles (arrow head) and significant lower quantities of vesicles. (e) At 23:30, external muscles in the control are completely destroyed, providing a non-occluded view of the internal oblique muscles (arrows). Mutant external muscles (arrow head) show a delayed onset of muscle degeneration. Internal muscles (arrows) remain mostly occluded. (f) At 33:00, apoptosis of persistent muscles is also completed in the mutant. Note that atrophy is more advanced in the control than in the mutant (f'), resulting in a smaller diameter of the muscle fibers (arrows). (g) At 77:00, persistent control muscles (arrow) show progressive erosion and an orientation parallel to the body axis. (g') Corresponding mutant fibers (arrow) display irregular morphologies and orientations. Note that the complete time series can be viewed as a movie (Additional File 1).