ZFIN ID: ZDB-FIG-130117-5
Bubenshchikova et al., 2012 - Wtip and Vangl2 are required for mitotic spindle orientation and cloaca morphogenesis. Biology Open   1(6):588-596 Full text @ Biol. Open
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: 1-cell to Long-pec

Fig. 1 Zebrafish wtip gene and protein during development.

(A) Zebrafish Wtip domain structure contains one nuclear export signal (NES, orange), one SH3 binding domain (SH3, green), three LIM domains (LIM1-3, yellow) and one PDZ binding domain (PDZ, red). (B) Percentage of amino acid identity between zebrafish Wtip and homologs of WTIP from other species. (C) RT-PCR time course of wtip expression during embryonic development. gapdh was used to control variations in total RNA (C,D). wtip expression was maternally deposited (C) and confirmed in the pronephros by RT-PCR using total RNA from isolated pronephros from 48 hpf embryos (D). (E) Cross-sections of the pronephric glomerulus and tubules. (F) Whole-mount in situ hybridization for wtip on embryos at 24 hpf, 34 hpf, and 48 hpf are side views anterior to the left and dorsal to the top. Zebrafish wtip expression is restricted to the eye, anterior brain, hindbrain, inner ear, and pronephros (E,F). Double immunofluorescence for the cilia marker acetylated α-tubulin (green) and Wtip (red) in confocal projections taken from whole-mount embryos for the middle segment of the pronephros at 24 hpf (G,H), the posterior pronephros at 24 hpf (I,J,K) and KV at the 10-somite stage (L,M,N). DAPI was used to counterstain nuclei (blue). Control embryos for antibody specificity using knockdown mediated by wtipMO of endogeneous Wtip protein translation (H,J,M) and with pre-immune serum (K,N) showed no expression. Localization of Wtip in the basal bodies of cilia was confirmed by double immunostaining with Wtip antibody (red) and either anti-γ-tubulin (P,V, green) or anti-centrin (S,Y, green), which are basal body markers in KV (V,Y, 10-somite stage) and the middle segment of the pronephros (P,S, 24 hpf). Pronephric tubules (pt), acetylated α-tubulin antibody (AT), γ-tubulin antibody (γ-tub), mid-pronephros (pk-mid), and posterior-pronephros (pk-post). Scale bars are 10 μm in E,G–N, 100 μm in F, and 2 μm in OZ.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
wtip AB standard conditions 1-cell to Long-pec whole organism RTPCR
Ab1-wtip 10-13 somites Kupffer's vesicle ciliary basal body IHC
Prim-5 brain anterior region ISH
Prim-5 eye ISH
Prim-5 heart tube ISH
Prim-5 hindbrain ISH
Prim-5 inner ear ISH
Prim-5 pronephros ISH
Ab1-wtip Prim-5 pronephros ciliary basal body IHC
Prim-15 brain anterior region ISH
Prim-15 eye ISH
Prim-15 heart tube ISH
Prim-15 hindbrain ISH
Prim-15 inner ear ISH
Prim-15 pronephros ISH
Long-pec brain anterior region ISH
Long-pec eye ISH
Long-pec heart ISH
Long-pec hindbrain ISH
Long-pec inner ear ISH
Long-pec pronephric glomerulus ISH
Long-pec pronephric tubule ISH
Long-pec pronephros RTPCR
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-cetn IHC AB standard conditions 10-13 somites Kupffer's vesicle ciliary basal body
IHC Prim-5 pronephros ciliary basal body
Ab1-tubg1 IHC AB standard conditions 10-13 somites Kupffer's vesicle ciliary basal body
IHC Prim-5 pronephros ciliary basal body
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Biology Open for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Biol. Open