piezo1 and piezo2a are essential for bone formation and mineral deposition in early stage of development. (A,B) Gross phenotypes and (C,D) mineralization patterns of sibling and piezo1−/−; piezo2a−/− mutant at 8 days after fertilization (dpf). White arrow indicates uninflated swim bladder. Scale = 500 μm. (E,F) Magnified images of the anterior body of sibling and double mutant. Numbers 1 to 8 indicate the position of each vertebral bone. The dashed circle indicates swim bladder. Notably, the swim bladder in double mutant was small and uninflated. (G) Schematic diagram of generating piezo transient KO mutant (CRISPRant). Comparison of 3D reconstruction of micro-CT images of vertebral bone at caudal part from (H) wildtype, (I)piezo1 sgRNA injected to piezo2a−/−, and (J)piezo2a sgRNA injected to piezo1−/− at 2.5 months after fertilization.
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