Figure 1: Activation of miniSOG in the epidermis causes paralysis and disrupts epidermal integrity.
From: Highly efficient optogenetic cell ablation in C. elegans using membrane-targeted miniSOG
(a) Cartoon of constructs targeting miniSOG to the outer membrane of mitochondria, and to cell membranes. (b) Representative images of adult animals immediately before and after blue light illumination. Blue light treatment results in paralysis of mito-miniSOG, myr-miniSOG, and PH-miniSOG expressing animals; times indicate the minimum time for immediate paralysis using 2 Hz blue light illumination. Scale, 250 μm. (c) Quantitation of paralysis immediately after blue light illumination, for the indicated times. Numbers are the animals that were analyzed in three independent experiments. (d). Quantitation of paralysis at different time points after 1 min blue light illumination. Paralyzed and non-paralyzed animals were counted at specific times after illumination. 4 independent experiments. Numbers are the animals that were analyzed. (e) Quantitation of locomotion velocity before and immediately after 1 min blue light illumination at 2 Hz. Transgenic animals were illuminated on 3 cm unseeded plates first and transferred to unseeded plates immediately afterwards for automated worm tracking. Numbers are the animals that were analyzed. (f) Representative DIC and confocal images of epidermis before and 4 h after 2 Hz blue light illumination. Images are from live, paralyzed animals expressing Pcol-19-PH-miniSOG. Left, DIC images, N indicates nuclei; arrows indicate vacuoles. Right, epidermal cells were labeled by Pcol-19-tdTomato. Yellow arrow indicates degenerated seam cell and white arrows indicate hyp7 after blue light illumination. Aggregation and decrease of the tdTomato fluorescence signal is evident. Red dash lines indicate epidermal edges. Scale, 10 μm. (g) Blue light illumination of PH-miniSOG in the epidermis increases lipid peroxidation. Representative confocal images of Bodipy C11 staining in lateral epidermis. Scale, 10 μm. (h) Quantitation of relative Bodipy C11 fluorescence intensity (AU). ***P < 0.001, Student’s t-test. Numbers are the animals that were analyzed.
