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ZFIN ID: ZDB-FIG-130426-18
Choe et al., 2013 - Wnt-Dependent Epithelial Transitions Drive Pharyngeal Pouch Formation. Developmental Cell   24(3):296-309 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Prim-15
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-15

Fig. 4 Wnt Signaling and Cdc42 Regulate Junctional Localization of Alcama in Developing Pouches(A–D) Immunohistochemistry shows progressive junctional localization of Alcama in the wild-type fourth pouch. Below each image, the normalized intensity of fluorescent signal is plotted along the 35 μM red lines. Asterisks indicate clear cell-cell boundaries.(E–H) Still images from a time-lapse confocal recording of fourth pouch development in nkx2.3:Alcama-GFP; her5:mCherryCAAX transgenic embryos (see Movie S6). As with endogenous Alcama protein, plots of fluorescence intensity through the red lines show progressive junctional localization of Alcama-GFP (green) within her5:mCherryCAAX-positive pouch cells (red).(I–X) Immunohistochemistry shows Alcama localization at the level of the fourth pouch in wild-type, mutant, and fzd8a-MO embryos, as well as UAS transgenic embryos (capitalized) doubly positive for nkx2.3:Gal4VP16. Alcama is partially mislocalized from pouch cell membranes to the cytoplasm in wnt4a-, wnt11r-wnt4a-, and fzd8a-MO embryos but not in wnt11r- embryos. In contrast, misexpression of Wnt11r results in disorganized endoderm (n = 28/32, data not shown) and occasionally rosette-like structures (n = 3/32), with rosette-like structures never observed in fzd8a-MO-injected siblings (n = 0/23, p = 0.003). Similarly, the inappropriate Alcama junctional localization induced by Wnt4a misexpression (n = 26/114) was never seen in fzd8a-MO-injected siblings (n = 0/74, p = 0.004) or doubly transgenic Wnt4a; DN-Cdc42 embryos (n = 0/31, p = 0.005). Rosette-like structures were observed in both CA-Rac1 embryos (n = 104/112) and their wnt11r mutant siblings (n = 11/17), and hyperelongated pouches were observed in both CA-Cdc42 embryos (n = 85/122) and their wnt4a-MO-injected siblings (n = 72/101). Plots of fluorescence intensity through the red lines are shown, with asterisks indicating clear cell-cell junctions.(Y–AB) Immunohistochemistry and normalized line plots show junctional localization of E-cadherin throughout wild-type pouch development, with pronounced apical enrichment (arrow in AA) as pouches mature. Inhibition of Dvl in nkx2.3:Gal4VP16; UAS:DvlΔDEP embryos does not significantly disrupt E-cadherin localization.See also Figure S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
alcama zn-8 WT standard conditions Prim-5 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 WT + MO2-fzd8a standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 el190Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 el268Tg; el48Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 el268Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 el320Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 el322Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 el322Tg; el93Tg + MO4-wnt4a standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 el421Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 el421Tg; el93Tg + MO2-fzd8a standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 el48Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 el48Tg; el93Tg + MO2-fzd8a standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 wnt11rfh224/fh224 standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 wnt11rfh224/fh224; el320Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 wnt11rfh224/fh224; wnt4afh295/fh295 standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
zn-8 wnt4afh295/fh295 standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction IHC
zn-8 Prim-15 pharyngeal pouch 4 cytoplasm IHC
EGFP el304Tg; el72Tg standard conditions Prim-5 pharyngeal pouch 4 cell-cell junction IFL
Prim-15 pharyngeal pouch 4 cell-cell junction IFL
mCherry el304Tg; el72Tg standard conditions Prim-5 pharyngeal pouch 4 plasma membrane IFL
Prim-15 pharyngeal pouch 4 plasma membrane IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-cdh1 IHC WT standard conditions Prim-5 pharyngeal pouch cell-cell junction
IHC Prim-15 pharyngeal pouch cell-cell junction
IHC el190Tg; el93Tg standard conditions Prim-15 pharyngeal pouch 4 cell-cell junction
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO2-fzd8a standard conditions Prim-15 pharyngeal pouch protein localization to cell junction occurrence, normal
Prim-15 pharyngeal pouch protein localization to plasma membrane decreased occurrence, abnormal
el190Tg; el93Tg standard conditions Prim-15 pharyngeal pouch protein localization to plasma membrane decreased occurrence, abnormal
el268Tg; el48Tg; el93Tg standard conditions Prim-15 pharyngeal pouch protein localization to cell junction occurrence, normal
el268Tg; el93Tg standard conditions Prim-15 pharyngeal pouch protein localization to plasma membrane decreased occurrence, abnormal
el320Tg; el93Tg standard conditions Prim-15 pharyngeal pouch endodermal cell irregular spatial pattern, abnormal
el322Tg; el93Tg standard conditions Prim-15 pharyngeal pouch endodermal cell elongated, abnormal
el322Tg; el93Tg + MO4-wnt4a standard conditions Prim-15 pharyngeal pouch endodermal cell elongated, abnormal
el421Tg; el93Tg standard conditions Prim-15 pharyngeal pouch endodermal cell irregular spatial pattern, abnormal
el48Tg; el93Tg standard conditions Prim-15 pharyngeal pouch protein localization to cell junction decreased occurrence, abnormal
wnt11rfh224/fh224 standard conditions Prim-15 pharyngeal pouch protein localization to plasma membrane occurrence, normal
wnt11rfh224/fh224; el320Tg; el93Tg standard conditions Prim-15 pharyngeal pouch endodermal cell irregular spatial pattern, abnormal
wnt11rfh224/fh224; wnt4afh295/fh295 standard conditions Prim-15 pharyngeal pouch protein localization to plasma membrane decreased occurrence, abnormal
wnt4afh295/fh295 standard conditions Prim-15 pharyngeal pouch protein localization to plasma membrane decreased occurrence, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 24(3), Choe, C.P., Collazo, A., Trinh, L.A., Pan, L., Moens, C.B., and Crump, J.G., Wnt-Dependent Epithelial Transitions Drive Pharyngeal Pouch Formation, 296-309, Copyright (2013) with permission from Elsevier. Full text @ Dev. Cell