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<italic>In vivo</italic> diffusion of Bdpy-ML in zebrafish larva.(A) Brightfield lateral view of a larva at 4 dpf with main anatomical structures highlighted. GI, Gastro-intestinal. (B) lateral views of 4dpf zebrafish Kdrl:Ras mCherry larvae, 2 and 24 hours post injection (hpi) in the caudal vein with vehicle (Veh), Sat-Bdpy-ML or Bdpy-ML. Images are maximal z projections of confocal tile scan images acquired on same larvae over time; mCherry fluorescence shown in red, Bdpy in green. Arrowheads indicate the spinal canal, arrows indicate the site of intravenous injection (iv), stars show accumulation in the GI tract at 24h. Areas delimited by dotted lines in (B) are shown at higher magnification in (C, 2 hpi) and (F, 24 hpi). Scale Bar 100 μm. (C) Enlarged views (with transmitted light image overlaid on fluorescence) of (B) showing accumulation of Bdpy-ML in heart and liver 2 hpi. (D) Detail of the vasculature in the ventral region of the tail showing longitudinal section of blood vessels and colocalization of Bdpy-ML with mCherry in endothelial cells of the caudal vein 4 hpi. N: notochord, cv (dotted brackets): caudal vein, da (brackets): dorsal aorta. (E) Dorsal view of the head with sections of blood vessels showing blood circulating Bdpy-ML and diffusion into the brain with accumulation in the tectal neuropil (tn) at 24 hpi. Right panel is an enlarged view a blood vessel in longitudinal section. (F) Enlarged view of the trunk in (B) showing preferential accumulation of Bdpy-ML in muscles (m), spinal canal (arrowheads) and spinal cord (double arrow) 24 hpi. N: notochord. (A, B, C, F): lateral views, dorsal top, anterior left; (D) lateral view, dorsal top right, anterior top left; (E) dorsal view, anterior down left. Scale bars (C-E) 40 μm. All images are representative of at least 10 larvae from 3 independent experiments. Images in (C-F) correspond to one focal plane (z = 2 μm).
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