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Fig. 3
The Notochord Affects KV Cell Shape by Polarizing Laminin Deposition around KV
(A) α-Laminin-α1β1γ1 antibody staining of KV in Tg(sox17:GFP) embryos at 1 ss (10.3 hpf; coronal section, A.1) and 6 ss (12 hpf; sagittal section, A.2 and A.3); A.4 and A.5 show insets of the region boxed in upper panels.
(B) Thickness of the KV epithelium [n = 42 measurements on 3 Tg(sox17:GFP) embryos]; box and whisker plot.
(C) Basal α-laminin-α1β1γ1 antibody staining intensity along KV epithelium in the same sections used for (B); n = 6 lines measurements; see dashed line in (A) for location; box and whisker plot; red line, trend based on the local average method.
(D) α-FN-1 antibody and DAPI staining on 6 ss (12 hpf) Tg(sox17:GFP) embryo (coronal section).
(E) α-Laminin-α1β1γ1 antibody and DAPI staining on ectopic KV in 6 ss (12 hpf) Tg(sox17:GFP) (left panel), MZoep; Tg(sox17:GFP) (middle panel), and MZoep; Tg(sox17:GFP) with a coinduced notochord (arrow in right panel) embryos.
(F) Ratio of nuclei densities between the AD and PV quadrants of KV as a function of average KV cell apical surface; mean ± SEM. Control n = 11 embryos; laminin γ1-MO1-cell n = 15 embryos; laminin γ1-MODFC n = 7 embryos; embryos injected with 40 pg truncated FN 1a and 1b mRNA n = 4 embryos. Data were polled from Tg(sox17:GFP) and Tg(sox17:LMA-tdTomato) embryos between 1 ss and 10 ss (10.3-14 hpf).
(G) Projected cilia length in control embryos and embryos injected with laminin γ1 MO either into the YSL between the 512- and 1,000-cell stages or at the 1-cell stage. p < 0.0001 (t test); n > 140 cilia for each condition; n > 5 embryos for each condition.
(H) Projected cilia length in control embryos and embryos injected with 40 pg truncated FN 1a and 1b mRNA; n > 89 cilia for each condition; n > 3 embryos for each condition; ns, p value (t test) > 0.05
(I) Quantification of the lateralization of spaw expression between 18 ss and 22 ss (20–22 hpf) by in situ hybridization in control (n = 18 embryos) and embryos injected with 40 pg truncated FN 1a and 1b mRNA (n = 36 embryos).
A, anterior; P, posterior; L, left; R, right; V, ventral; D, dorsal; ns, not significant; YSL, yolk syncytial layer; ctrl, control; trunc. fibro., embryos injected with 40 pg truncated FN 1a and 1b mRNA. Scale bars in (A), (D), and (E), 20 µm. See also Figure S3.
Reprinted from Developmental Cell, 31, Compagnon, J., Barone, V., Rajshekar, S., Kottmeier, R., Pranjic-Ferscha, K., Behrndt, M., Heisenberg, C., The Notochord Breaks Bilateral Symmetry by Controlling Cell Shapes in the Zebrafish Laterality Organ, 774-783, Copyright (2014) with permission from Elsevier. Full text @ Dev. Cell