Comparison of Plasma Membrane Distribution of Rab5c-YFP and Number of Cell Nuclei during Early Epiboly
(Related to Figure 3)
(A-F) Confocal anti-GFP immunofluorescence images of Rab5c-YFP expressed by mRNA injection in WT (A-B) or MZspg embryos (C-D), and of Rab5c-YFP and RN-tre co-expressed in WT (E-F) embryos by mRNA co-injection. Animal views. Scale bar = 10 μm.
(G-H) Quantification of cell nuclei number and thickness of deep cell layer (DCL) from two hour (sphere to 50% epiboly) 3D time-lapse recordings of standard control morpholino (Con Mo) injected (n= 6 embryos), mgfp mRNA injected (n= 7 embryos), Rab5c morpholino (Rab5c Mo) injected (n= 7 embryos), or RN-tre mRNA injected (n= 5 embryos) WT embryos in which all cell nuclei were labeled by expression of NLS-tomato. Each embryo is documented by time series of animal pole view confocal stacks 120 μm from the animal pole EVL into the blastoderm. Cell nuclei numbers from the 3D time-lapse volume data in Rab5c Mo injected embryos (G) or RN-tre mRNA injected embryos (H) are not significantly different from control WT embryos, while thickness of the DCLs at 5.5 hpf as percentage 8 of DCL thickness at 4 hpf are significantly thicker both in Rab5c Mo (p < 0.05) and RN-tre mRNA (p < 0.01) injected embryos.
Boxplots and trend lines represent cell nuclei number and DCL thickness, respectively. Error bars represent standard error of the mean (SEM).
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Reprinted from Developmental Cell, 24(5), Song, S., Eckerle, S., Onichtchouk, D., Marrs, J.A., Nitschke, R., and Driever, W., Pou5f1-dependent EGF expression controls e-cadherin endocytosis, cell adhesion, and zebrafish epiboly movements, 486-501, Copyright
(2013) with permission from Elsevier.
Full text @ Dev. Cell