Fig. 2. Loss of ATP4 causes defects in MCCs reminiscent of foxj1 and Wnt/PCP phenotypes. (A–C) Analysis of the cellular phenotype in MCCs using high magnification confocal single cell imaging. (A) Cells were stained with an antibody against acetylated-α-tubulin (tubulin, red) and phalloidin-Alexa488 for actin staining (actin, green). Control (uninj.) MCCs were characterized by the presence of a dense ciliary tuft projecting from the apical surface (dashed lines in lateral projections) and the presence of an apical actin meshwork. In contrast, MCCs in atp4a morphants showed reduced ciliation, intracellular accumulation of tubulin and defects in the apical actin meshwork formation. (B) The localization of basal bodies to the apical membrane was analyzed using overexpression of sas6-gfp (green) in combination with tubulin (blue) staining, the outlines of the cells are depicted by the dashed lines in the apical views. Control MCCs were fully ciliated and basal bodies aligned close to the apical membrane (dashed lines in lateral projections), while atp4a morphant MCCs had severe defects in ciliation, which correlated with aberrant basal body distribution within the cell. Basal bodies in atp4a morphants were mislocalized to the deep cytoplasm and not uniformly distributed along the apical membrane. (C) Basal body orientation was analyzed using centrin4-rfp (red) and clamp-gfp (green) overexpression. In control MCCs, basal bodies were mostly uniformly aligned along the anterior–posterior axis, while this alignment was randomized in atp4a morphant embryos. Lower panels show a magnified field of basal bodies.
Image published in: Walentek P et al. (2015)
Copyright © 2015. Image reproduced with permission of the Publisher, Elsevier B. V.
| Gene | Synonyms | Species | Stage(s) | Tissue |
|---|---|---|---|---|
| tuba4b.L | alpha tubulin, alpha-tubulin, tuba4, tuba4a | X. laevis | Throughout NF stage 15 | ciliated epidermal cell |
Image source: Published
| Experiment + Assay | Source | Phenotypes and Disease | ||||
|---|---|---|---|---|---|---|
| Xla Wt + atp4a MO + NF32 (immunohistochemistry) | Fig. 2 A |
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