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Figure Lengend Snippet: a, Time-course analysis of E-cadherin and N-cadherin expression during neural tube formation. E- cadherin was initially broadly distributed but decreased in cells associated with the Matrigel region, whereas N-cadherin expression was initially weak and became apparent in cells on the Matrigel from day 6 onward. By day 10, the initially continuous monolayer had segregated into two distinct tissues corresponding to the surface ectoderm and neural tube. Scale bar, 50 μm. b, Three-dimensional imaging of tissues stained for N-cadherin, E-cadherin and cytokeratin 8. Section A–A′ shows an incompletely closed region, whereas section B–B′ shows a fully closed neural tube. Scale bar, 70 μm. c, Time-lapse imaging of actin dynamics from day 9 of culture (0 h) to day 10 (24 h). White arrows indicate actin-rich structures bridging the opposing tissue edges during closure. Scale bar, 30 μm. d, Myosin II distribution in the cell sheet during neural induction. Myosin II accumulated at the apical side of the cell sheet within the Matrigel-associated region. e, Distribution of SOX10 and ZIC1 before neural tube closure. Both markers were enriched at the boundary region between the neural and non- neural domains.

Article Snippet: and Supplementary Video 4 show two-photon time-lapse imaging from day 9 to day 10, allowing visualization of actin dynamics by using SPY-actin (CY-SC202, Cytoskeleton Inc.) during surface ectoderm closure.

Techniques: Analysis, Expressing, Imaging, Staining, Tissue