XB-IMG-146168
Xenbase Image ID: 146168
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Fig. 3. Ezrin shows expression in cilia and is required for ciliogenesis in zebrafish. (A-D) Human respiratory epithelial cells from healthy controls were double labelled with antibodies directed against Ezrin (green) and the ciliary axoneme marker α/β-Tubulin (magenta) (arrow). Ezrin localisation is restricted to the putative basal bodies (arrowhead) and the nucleus. The nucleus is stained with Hoechst 33342 (blue). (E-G) Ezrin is expressed at the basal bodies (arrowheads) and the ciliary axonemes in 48 hpf Tg(actb2:Mmu.Arl13b-GFP) zebrafish embryos. (H-I′) Expression silencing of ezrin (I-I′) using TB-MO ezrin (2 ng) results in hydrocephalus (arrowhead in I) and pronephric cyst formation (stars in I′ and I′) as compared with zebrafish embryos injected with Co-MO (2 ng) (H-H′), shown in a bright-field lateral view with anterior to the left (H,I), a dorsal view with anterior to the left of a Tg(wt1b:EGFP) embryo (H′,I′), and by a histological transverse section (H′,I′) of 48 hpf embryos. (J) Quantification of pronephric cyst formation in 48 hpf zebrafish embryos after injection with TB-MO ezrin (2 ng) or TB-MO ezrin (2 ng)+ ezrin mRNA (20 pg), as compared with Co-MO (2 ng). There is significant prevention of cyst formation upon co-injection of ezrin mRNA (***P<0.001). (K,K′) TEM analysis revealed reduced microvilli formation and basal body docking defects in TB-MO ezrin (2 ng) morphants at 48 hpf. Arrow indicates prospective basal body not properly docked. (L) Quantification of pronephric cyst formation in 48 hpf zebrafish embryos injected with Co-MO (2 ng), TB-MO ezrin (2 ng), TB-MO ezrin (2 ng)+ elmo1 mRNA (20 pg), SB-MO elmo1 (2 ng) or SB-MO elmo1 (2 ng)+ ezrin mRNA (20 pg) (*P=0.03; **P=0.006). (J,L) The number of individual embryos analysed is indicated above each bar. Scale bars: 10 µm in D; 5 µm in G; 100 μm in H,I; 50 µm in H′,H′,I′,I′; 2 μm in K; 0.5 μm in K′. Image published in: Epting D et al. (2015) Copyright © 2015. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |