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ZFIN ID: ZDB-FIG-180822-3
Guo et al., 2018 - Apical Cell-Cell Adhesions Reconcile Symmetry and Asymmetry in Zebrafish Neurulation. iScience   3:63-85 Full text @ iScience
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Condition:
Anatomical Terms:
Stage Range: 14-19 somites to 26+ somites

Fig. 2

Dynamics and Compositions of the PAAs and OAAs

(A) Diagrams illustrate the principle of GFP-assisted sagittal serial microscopy for visualizing proteins at the opposing apical surfaces: the opposing apical surfaces at the midline region (SM), which contain the OAAs, must be flanked by two spatial references—PAA regions of the left and right halves of the tissue (SL and SR). Green represents transiently expressed GFP in some cells. The bottom panels are images collected with this technique: N-Cad and β-catenin were detected at the PAA regions as circumferential belts (SL and SR, arrows) as well as in the OAA regions as punctate foci (SM; arrowheads). By contrast, ZO-1 and actin were detected in the PAA regions (SL and SR, arrows) but not in the OAA regions (SM, arrowheads).

(B) Diagrams illustrate the strategy to visualize transiently expressed N-Cadwt-GFP or N-Cadm117-GFP at 14-ss. Note that N-Cadwt-GFP enriched on the opposing apical surfaces (illustrated by the drawing on the right).

(C) The PAAs (arrows) and the opposing apical surfaces (arrowheads indicating the OAAs) survived the Triton X-100 extraction of the anti-GFP immunogold electron microscopic procedure, whereas the lateral membranes were dissolved (basal of asterisks).

(D) The magnification of the boxed region in C revealed the labeling of N-Cad-GFP with 5-nm gold particles at the PAAs (arrows, 72% of the PAAs were labeled with gold particles) and OAAs (arrowhead). Insets are magnifications of boxed regions.

(E) Crb2a juxtaposed with ZO-1 sites at the apical ends (26-ss).

(F) The table summarizes the developmental and compositional changes of the OAAs and PAAs. Crb*, Crb-based adhesion; NaKATPase*, Na+/K+-ATPase-based adhesion; +, presence; -, absence. Also see Figure S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP pt137Tg heat shock 14-19 somites neural tube cell-cell adherens junction IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-cdh2 IHC TU standard conditions 14-19 somites neural rod apical plasma membrane
IHC 14-19 somites neural tube cell-cell adherens junction
Ab1-crb2a IHC TU standard conditions 26+ somites neural tube apical plasma membrane
IHC 26+ somites neural tube cell-cell adherens junction
Ab1-ctnnb IHC TU standard conditions 14-19 somites neural rod membrane
Ab1-tjp1 IHC TU standard conditions 14-19 somites neural rod apical plasma membrane
IHC 14-19 somites neural tube cell-cell adherens junction
IHC 26+ somites neural tube apical plasma membrane
IHC 26+ somites neural tube cell-cell adherens junction
IHC pt137Tg heat shock 14-19 somites neural tube cell-cell adherens junction
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal iScience for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ iScience