![]() ( A) Quantification of KRT8+ cells per FFP section reveals taste bud cell number does not increase until P9. Blue bars: mean ± SD (n = 3 animals per stage, 10 placodes per animal, open and shaded shapes). ( P, Q) Total cell number and placode volume did not differ between stages. White outlines ( L, N) and randomly assigned colors ( M, O) indicate individual placode cells. ( L–O) E13.5 and E16.5 taste placodes (KRT8+, magenta) in E-cadherin (Ecad) immunostained whole tongues were imaged through their apical-to-basal extent, 3D reconstructed and individual cells defined in Imaris (see Materials and methods). Blue bars: mean ± SD (n = 3 animals per stage, 6–11 placodes per animal, open and shaded shapes). ( A– J are confocal optical sections acquired at 0.75 µm.) ( K) Quantification of placodal KRT14-Tomato+ cells 48 hr after tamoxifen induction at progressive stages, for example tamoxifen at E12.5, analysis at E14.5. ( J) Krt14 CreERT2 R26R tdTomato (KRT14-tomato green) induced at E12.5 labels a subset of KRT8+ (magenta) placode cells at E14.5. ( G–I) At E16.5, KRT8 is expressed exclusively by taste bud primordia ( H), which lack KRT14, while non-taste basal epithelial cells are robustly KRT14+. KRT14 is evident only basally and apically in placodes (asterisk in F) but remains well expressed in non-taste epithelium. ( D–F) At E13.5, KRT8 expression is expressed by taste placodes (asterisk in F), and surface periderm ( E, yellow arrows). ( A–C’) Before taste placodes are evident (E12.0), KRT14 (green) and KRT8 (magenta) are co-expressed in lingual epithelium.
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