(A) When the fluorogen MG2I (chemical structure shown on left) is bound to the fluorogen-activating protein (FAP) dL5** (right), excitation by far-red light causes generation of singlet oxygen. (B) …
(A) A LED light source (100W, 660 nm; Chanzon, Amazon.com) was mounted on a CPU heatsink and fan for cooling. The LED/fan assembly was suspended beneath an optical glass window on a custom stand, so …
(A) A waterproof resin-coated negative temperature coefficient thermistor was used to provide continuous monitoring of bath temperature. The resistance of the thermistor was determined in a voltage …
(A) Design of experiments shown in panels (B - D). Experimental groups: WT zebrafish (black); WT zebrafish exposed to MG2I (green); NeuMitoFAP zebrafish (blue); NeuMitoFAP zebrafish exposed to MG2I …
Source data for Figure 2C.
Source data for Figure 2E.
Transmitted light micrographs of NeuMitoFAP zebrafish: tThe right column shows larvae that were exposed to MG2I from 3 dpf; the left column shows untreated zebrafish. The first row shows larvae at …
(A) Visual motor responses were evaluated in Tg(eno2:gal4FF); Tg(UAS:COX4-COX8-dL5-mCer3) (‘NeuMitoFAP’) zebrafish treated with MG2I. Motor response to cycles of green light (200 Lux; 10 min) and …
(A) Visual motor responses were evaluated in NeuMitoFAP (red) and non-transgenic (green) zebrafish treated with MG2I, identically to Figure 2—figure supplement 1. Each group of 12 zebrafish was …
24 zebrafish from each experimental group (non-transgenic and NeuMitoFAP zebrafish, not exposed to chemical or treated with MG2I; same color key as Figure 2B) were placed in a 96 well plate at 5dpf. …
(A) Whole-cell patch clamp recordings were made from posterior lateral line ganglion (PLLG) sensory neurons. The inset figure shows the preparation and experimental design (the arrow shows a …
Source data for Figure 3B-D.
The graphs show membrane potential values (y-axis) of lateral line ganglion sensory neurons over time (x-axis) in whole cell patch clamp recordings from multiple zebrafish in each experimental …
Whole-cell patch clamp recordings from lateral line ganglion neurons were made exactly as in Figure 4, with the exception that illumination at λpeak=661nm was stopped after 5 min (black; 13.5 J/cm2) …
(A) ATP concentration was measured in lysates (each containing 5 whole zebrafish larvae) using a bioluminescent assay and normalized to protein content. Data points show ATP/protein in individual …
Source data for Figure 4A.
Source data for Figure 4B.
Source data for Figure 4D-E.
Source data for Figure 4F.
It was necessary to prevent swimming movements during oximetry measurements (Figure 4) because: (i) movements of larvae could disrupt measurement of dissolved O2 in the water; and (ii) the large …
Bath acidification rate (BAR) was quantified at the same time as the oxygen consumption rate measurements shown in Figure 4, using a Seahorse XF24 flux analyzer. (A) In the first 30 min of …
(A) Confocal Z-plane projections showing mCerulean-labeled mitochondria in the lateral line nerves of live NeuMitoFAP zebrafish in the absence (upper images) or presence (lower images) of MG2I, …
Source data for Figure 5B.
Source data for Figure 5C.
Source data for Figure 5D.
Source data for Figure 5G.
NeuMitoFAP zebrafish larvae were treated with MG2I or no chemical from 3 to 5 dpf and then exposed to 60 J/cm2 light at λpeak=661 nm, following which they were fixed immediately for electron …
(A) Mean swimming speed of NeuMitoFAP zebrafish treated with MG2I was quantified during the dark phase of the visual motor response (y-axis) after exposure to 60 J/cm2 far-red light (red) or no …
Source data for Figure 6D.
5dpf zebrafish swimming in the wells of an agarose-filled plate were illuminated from above by an infrared light, and video was recorded from below at 30 frame/s, during the dark phase of the visual …
(i) Phase contrast videomicrography of 5dpf zebrafish from the same experimental groups as Video 1, illustrating identical circulation of blood cells in the vascular system from all experimental …
7dpf zebrafish are shown 48 hr after far-red light exposure at 5dpf. Methods and experimental groups are identical to Video 1. The severe neurological abnormalities seen immediately after far-red …
Phase contrast videomicrography of 7dpf zebrafish 48 hr after far-red light exposure at 5dpf. Methods and experimental groups are identical to Video 2. Despite the persistence of severe neurological …
Red | Green | IR | dL5**-MG2I (minor) | dL5**-MG2I (major) | |
---|---|---|---|---|---|
Peak λ (nm) | 661 | 516 | 877 | 456 | 666 |
Centroid λ (nm) | 656 | 520 | 880 | 452 | 649 |
FWHH (nm) | 18 | 36 | 51 | 55 | 61 |
Red x dL5**-MG2I | Green x dL5**-MG2I | |
---|---|---|
Peak λ (nm) | 661 | 524 |
Height | 0.98 | 0.013 |
Area | 19.10 | 1.15 |