ZFIN Image: Verweij et al., 2019, Fig. 7

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Figure Caption

Fig. 7

Functional Analysis of YSL-Exosomes

(A) Transmission and fluorescent images of Ctrl MO- or SyntA MO-treated 1 dpf zebrafish embryos.

(B) Quantification of morphological features of Ctrl MO- or SyntA MO-treated 1–3 dpf (24–72 h) zebrafish embryos. To the left, hatched versus non-hatched comparison. To the right, health status comparison with normal morphology (healthy), mild or severe developmental defects, and mortality. Sample size ≥ 120.

(C) Size (length) of Ctrl MO- or SyntA MO-treated 3 dpf zebrafish embryos (scatter-dot plot, mean ± SD; n = 19, n = 18, respectively).

(D) Close up of the CVP area of 3 dpf Tg(kdrl:Hsa.HRAS-mCherry) zebrafish embryos expressing CD63-pHluorin in the YSL, co-injected with Ctrl MO or SyntA MO in the YSL. Left panel: frontal view; right panel: lateral view. CA, caudal artery; CV, cardinal vein.

(E) Fluorescent intensity of GFP signal in CA measured during different treatments. BafA, bafilomycin-A; Pyr, Pyrimidyn-7; DS500K, 500 KDa dextran sulfate; SyntA MO, syntenin-a morpholino; A.U., arbitrary units. mean ± SD; ^∗∗∗∗p ≤ 0.0001; all conditions show at least triplicates of n ≥ 3; unpaired t test with equal SD.

(F) Quantification of overlap-coefficient of red with green (M1) and green with red (M2) in Ctrl MO- or SyntA MO-treated 3 dpf Tg(kdrl:Hsa.HRAS-mCherry) zebrafish embryos, as shown in (D) (mean ± SD; ^∗∗∗∗p ≤ 0.0001; n = 5; unpaired t test with equal SD).

(G and H) Quantification of height of caudal part of vasculature, specifically for the CVP at second intersegmental vessel (ISV), or (H) measured from CA to CV (scatter-dot plot, mean ± SD; ^∗p ≤ 0.05; n ≥ 10, n ≥ 9 resp; unpaired t test with equal SD).

(I–K) Tg(kdrl:eGFP /nls-mKate2) embryos injected with control (Ctrl MO) or syntenin-a (SyntA MO) morpholinos in the YSL, analyzed for growth (I), proliferation (J), and nuclear density (K) in the CVP between 35 and 40 hpf (mean ± SD; ^∗p ≤ 0.05; ^∗∗p ≤ 0.01; n = 4; unpaired t test with equal SD).

(L) Time-lapse of Tg(kdrl:eGFP /nls-mKate2) embryos between 35 and 40 hpf. Boxes indicate 2 examples of proliferating cells. as shown in (M). Scale bar represents 40 μm.

(M) Two examples of dividing cells are shown. Scale bar represents 15 μm. YSL, yolk syncytial layer; CVP, caudal vein plexus.

Figure Data

Phenotype details

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image.

Reprinted from Developmental Cell, 48(4), Verweij, F.J., Revenu, C., Arras, G., Dingli, F., Loew, D., Pegtel, M.D., Follain, G., Allio, G., Goetz, J.G., Zimmermann, P., Herbomel, P., Del Bene, F., Raposo, G., van Niel, G., Live Tracking of Inter-organ Communication by Endogenous Exosomes In Vivo, 573-589.e4, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell