DEV02678 Supplementary Material

Files in this Data Supplement:

• Supplemental Table 1 - Adobe PDF

• Supplemental Figure 1 -

  Fig. S1. Initiation of keel formation relative to eye-field location in wild-type, silberblick and cyclops embryos. (A-C) Wild type. (D-F) slb. (G-I) cyc morphant. (A-I) Dorsal views, anterior to left. (A,D,G) Forebrain regions, defined by cell tracking, at 80% epiboly (8.4 hpf). White cells and open circles − medial anterior eye (left) and initiation of keel formation (right). Adjoining solid line − intervening distance along the midline (square dotted line). Keel formation begins with hypothalamus in wild-type (A) and slb (D), and with anterior eye field in cyc morphant (G). For corresponding angular distances subtended at the embryonic centroid see Table S1. (B,E,H) Odd-paired-like (opl) gene expression, as determined using standard procedures, defines the eye field at 80% epiboly (8.4 hpf). Hypothalamus occupies the posterior-medial notch in wild type (B) and slb (E), consistent with the tracked data in A and D. However, eye tissue presides in the posterior-medial notch in cyc morphant (H). (C,F,I, Table S1) DAPI nuclear counter-staining, defining the medial boundaries (red cells and open circles) of the opl-expressing eye field (dashed line) at 80% epiboly (8.4 hpf), confirms this. hpf, hours post fertilisation. Scale bars: 25 μm.

• Supplemental Figure 2 -

  Fig. S2. The resolution of bilateral eyes during forebrain neurulation. (A-D,I-L) Projection images of GFP-labelled nuclei. (E-H,M-P) 3D projections of tracked cells. Arrows indicate the direction of major tissue movement. Dotted line, neural boundary; dashed lines, dorsal midline, neuropore. (A-D,I-L) Dorsal views, anterior is left; (E-H, M-P) frontal views, dorsal is up. (A,E) Forebrain regions prior to neurulation. Cells of the left and right eyes are intermixed at the midline. Left and right eye-field cells remain intermixed throughout the period of posterior contraction of the anterior neural plate (B,F) and throughout subduction of the hypothalamus (asterisk) beneath the medial eye field (C,D,G,H). (I, M) Sorting of eye tissue into the appropriate vesicles begins with the movement of cells away from the midline during evagination, (J-L, N-P) and continues coincident with the closure by the telencephalon of the neuropore, and the convergence and anterior-ward movement of the dorsal diencephalon. hpf, hours post fertilisation. Scale bars: 25 μm;

• Movie 1 -

  Movie S1. Time-lapse analysis of eye-field morphogenesis during forebrain neurulation. 3D projection of GFP-labelled nuclei, highlighting the individual cell movements followed during forebrain formation and the resolution of bilateral eyes. Dorsal view, anterior is left. Horizontal white line, dorsal midline; curved white line, neuropore; asterisk, initiation of keel formation. The timing of the principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.

• Movie 2 -

  Movie S2. 3D projection of tracked cells enacting forebrain neurulation. Frontal view, dorsal is up. Vertical white line, dorsal midline; asterisk, initiation of keel formation. The timing of the principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.

• Movie 3 -

  Movie S3. 3D projection of the tracked cells enacting eye-field morphogenesis. A latero-medial slice highlighting the reorganisation of the eye field during forebrain neurulation. Frontal view, dorsal is up. The timing of principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.

• Movie 4 -

  Movie S4. Animation illustrating the principal folding movements of the forebrain. View is of the complete width of the forebrain, observed dorsally. This animation spans development from neural-plate contraction to closure of the neural pore. The spherical curvature of the zebrafish embryo has not been represented and some exaggeration in the depth of the tissue has been introduced to aid visualisation. Anterior is left.

• Movie 5 -

  Movie S5. Animation illustrating the principal folding movements of the forebrain. View is of the complete width of the forebrain, observed obliquely from the left. Anterior is bottom-left and dorsal is up.

• Movie 6 -

  Movie S6. Virtual tour illustrating the principal folding movements of the forebrain. Drag the image from left to right to develop in time; drag the image up and down to tilt the neural plate.

• Movie 7 -

  Movie S7. Time-lapse analysis of eye-field morphogenesis in silberblick. 3D projection of GFP-labelled nuclei, showing that reduced cell movements during medial eye field reorganisation underlies defective evagination. Dorsal view, anterior is left. Horizontal white line, dorsal midline; curved white line, neuropore; asterisk, initiation of keel formation. The timing of the principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.

• Movie 8 -

  Movie S8. 3D-Projection of tracked cells enacting eye field morphogenesis in silberblick.Frontal view, dorsal is up. Vertical white line, dorsal midline; asterisk, initiation of keel formation. The timing of the principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.

• Movie 9 -

  Movie S9. Time-lapse analysis of eye-field morphogenesis in cyc MO. 3D projection of GFP-labelled nuclei, showing that the erroneous subduction of eye tissue precludes eye-field splitting by mechanisms seen in wild-type animals. Dorsal view, anterior is left. Horizontal white line, dorsal midline; curved white line, neuropore; asterisk, initiation of keel formation. The timing of the principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.

• Movie 10 -

  Movie S10. 3D-projection of tracked cells enacting eye-field morphogenesis in cyc MO. Frontal view, dorsal is up. Vertical white line, dorsal midline; asterisk, initiation of keel formation. The timing of the principal morphogenetic phases is shown. Time, in hours post fertilisation. Scale bar: 25 mm.