Fig. 5: 3D morphometrics to identify cell behaviors in basement membrane-depleted embryos.
From: Basement membrane perforations guide anterior–posterior axis formation
a Schematic of an E5.5 embryo showing the relationship between major cell types: EPI (yellow), VE (white), DVE (green), EPC (gray), and ExE (cyan). b Confocal image of a fixed E5.5 E-cadherin-GFP (Cdh1-GFP in gray) embryo with CERL immunostained (green) to identify the migrating DVE. Scale bars:20 μm. c In toto imaging of a fixed E5.5 embryo using multiview light-sheet microscopy. Embryos rotated at 3 different angles to comprehensively show VE. d VE segmented, 3D reconstructed, and color-coded embryo rotated at 4 different angles. e–g Morphometric analyses of E5.0, E5.25, and E5.5 embryos: the volume of the entire embryo (e), aspect ratio (f), and cell number (g). n = 8, 18, and 8 embryos. h Extraction of tissue topology from 3D reconstructed and VE segmented embryo. Cell sidedness is color-coded. i, j Comparison of morphometrics in acute collagenase-treated E5.5 Cdh1-GFP embryos cultured for 3 h and the control. While the number of VE cells remains similar (i), the percentage of hexagonal VE cells is lower in basement membrane-disrupted embryos than in the controls (j). n = 15 and 16 embryos. p-values = 0.22(n.s., not significant) and 0.02(*); two-sided Student’s t test. Error bars: standard deviation.
