Figure 3: Membrane targeting enhances the efficiency of miniSOG mediated GABAergic, touch, and interneuron ablation.
From: Highly efficient optogenetic cell ablation in C. elegans using membrane-targeted miniSOG
(a) Punc-25-PH-miniSOG transgenic animals display Unc phenotypes immediately after 4 min blue light illumination, whereas Punc-25-mito-miniSOG animals did not. Scale, 250 μm. (b) Quantitation of locomotion speed immediately after blue light illumination. Punc-25-PH-miniSOG transgenic animals displayed significantly reduced speed compared to Punc-25-mito-miniSOG. Numbers are the animals analyzed in 2 independent experiments. ***P < 0.001, t-test. (c) Quantitation of shrinker phenotype in Punc-25-mito-miniSOG and Punc-25-PH-miniSOG expressing animals, before and after blue light illumination (4 min at 2 Hz). n > 30 animals for each transgenic animal. ***P < 0.001, t-test. (d) Representative confocal images of PLM neurons before and 24 h after 4 min blue light illumination. The PLM neuron is degenerated in mito-miniSOG and in PH-miniSOG transgenic animals after illumination. Arrows indicate PLM processes. Scale, 20 μm. (e) Quantitation of touch responses in Pmec-4-mito-miniSOG and Pmec-4-PH-miniSOG before and after blue light illumination. ***P < 0.001, t-test. (f) Quantitation of backward movement after head touch in Pnmr-1-mito-miniSOG and Pnmr-1-PH-miniSOG transgenic animals after blue light illumination. ***P < 0.001, t-test. (g) Representative confocal images of PVC neurons before and 24 h after 4 min blue light illumination. PVC neuron is degenerated in mito-miniSOG and PH-miniSOG transgenic animals after illumination. White arrow indicates PVC neurons and yellow arrows indicate mCherry debris in the epidermis. Scale, 10 μm.
