ZFIN ID: ZDB-FIG-120423-45
Boggetti et al., 2012 - NBP, a zebrafish homolog of human Kank3, is a novel Numb interactor essential for epidermal integrity and neurulation. Developmental Biology   365(1):164-174 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-5

Fig. 3

NBP and Numbs morphants show defects in development. Panels compare NBP morphants (NBP MO), NBP control morphants (NBP MOC) and Numbs morphants (Numbs MO) with wild-type embryos (WT). (A) 24 hpf embryos injected with either NBP MO or Numbs MO are delayed in growth and abnormally developed in comparison with wild-type embryos.Arrows point to dissociated cells in the dorsal region of the NBP morphants (lateral view, anterior towards the left). (B) Western blot analysis of protein extracts from 24hpf wild-type and NBP MO-injected embryos with the N1 polyclonal antibody against NBP. The wild-type extract gives a NBP-positive band of the expected size (99.57 kDa), whereas only a negligible amount of NBP is seen in extracts from the morphants. (C, E–H) Anomalous development of the epidermis. The epidermal cells of the NBP MO-injected embryos varied in size and shape, were separated by large intercellular spaces (C) and frequently were released from the surface (E, asterisk denotes the gap below the epidermal cells; arrowheads in F and H). Tumor-like structures emerged from the dorsal part of the embryo (asterisks in F, G, and H). They were composed of enlarged cells with round, β-catenin strongly positive nuclei (F) and with α-catenin (G), β-catenin (F) and E-cadherin (H) enriched in the cell–cell contacts. Stars point to inner tissue located just underneath the tumor-like structures where cells lack β-catenin in both nuclei and membranes (F) and α-catenin in membranes (G). Arrows point to junctions between neighboringepidermal cells enriched in β-catenin (F), α-catenin (G) and E-cadherin (H) in WT and to mislocalized E-cadherin in epidermal cells of NBP MO (H). Nuclei in H are stained by acridine orange. Scale bar: A, D = 100 μm, C, F–H = 20 μm, and E = 1 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
kank3 Ab1-kank3 WT standard conditions Prim-5 whole organism WB
Ab1-kank3 WT + MO1-kank3 standard conditions Prim-5 whole organism WB
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-cdh1 IHC WT control Prim-5 epidermal cell cell-cell junction
IHC Prim-5 epidermis
IHC WT + MO1-kank3 standard conditions Prim-5 epidermal cell cell-cell junction
IHC Prim-5 epidermis
Ab2-ctnna IHC WT control Prim-5 epidermal cell cell-cell junction
IHC Prim-5 epidermis
IHC WT + MO1-kank3 standard conditions Prim-5 epidermal cell cell-cell junction
IHC Prim-5 epidermis
Ab6-ctnnb IHC WT control Prim-5 epidermal cell cell-cell junction
IHC Prim-5 epidermal cell nucleus
IHC Prim-5 epidermis
IHC WT + MO1-kank3 standard conditions Prim-5 epidermal cell cell-cell junction
IHC Prim-5 epidermal cell nucleus
IHC Prim-5 epidermis
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-kank3 standard conditions Prim-5 cell-cell adhesion decreased process quality, abnormal
Prim-5 chordate embryonic development delayed, abnormal
Prim-5 epidermal cell increased variability of size, abnormal
Prim-5 epidermis epidermal cell separated from epidermis epidermal cell, abnormal
Prim-5 epidermis morphogenesis decreased process quality, abnormal
Prim-5 whole organism malformed, abnormal
WT + MO3-numb standard conditions Prim-5 chordate embryonic development delayed, abnormal
Prim-5 whole organism malformed, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 365(1), Boggetti, B., Jasik, J., Takamiya, M., Strähle, U., Reugels, A.M., and Campos-Ortega, J.A., NBP, a zebrafish homolog of human Kank3, is a novel Numb interactor essential for epidermal integrity and neurulation, 164-174, Copyright (2012) with permission from Elsevier. Full text @ Dev. Biol.