Anillin expression during zebrafish retinal differentiation. (A) anillin mRNA (blue) at 28 hpf. (B) Expression of the anillin-eGFP transgene. R, retina; T, optic tectum; L, lens. Left, z-projection view; middle and right, z-plane. (C,C2) anillin:anillin-eGFP/ath5:gap43-RFP transgenic retina. anillin-eGFP is downregulated in the differentiating cells of the RGC layer (asterisks). (D) Anillin-GFP localisation in the nucleus (asterisk), cleavage furrow (arrow, t=02) and at the midbody (arrow, t=52 and t=10′). Time, minutes. (E,E′) Anillin-eGFP symmetric (E) and asymmetric (E′) distribution and intensity profiles (data points and fitted Gaussian) used to detect the offset of the Anillin-eGFP spot from the daughter cell boundary. (F) The position of apical Anillin-eGFP (n=15 divisions). Distance from the cell boundary within the average radius of the Anillin spot (grey) is used to define symmetry (see supplementary material Fig. S3). (G) Frames from supplementary material Movie 1. The Anillin-eGFP spot-inheriting daughter (white arrowhead, red dot) migrates to the RGC layer. The sibling (blue dot) migrates back to the apical surface (n=5 out of 5 analysed divisions). At t=20′ a rotated frontal z-section (dotted lines, 3D slicing mode) across the centre of the dividing daughters is shown (oriented along the z-plane). The apical surface of the retinal neuroepithelium is to the top. Scale bars: 110µm in B; 25µm in C,C′; 6µm in E,E′; 12µm in G.
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