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rspo3 ablation does not impact initial bone mineralization but increases osteoclast area of activity. (A) Representative images of maximum intensity projections of confocal z-stack of whole-mount 10 dpf zebrafish. Zebrafish express sox10:kaede transgene allowing fluorescent imaging of cartilage elements. Zebrafish were live-stained with Alizarin red S to visualize mineralized structures. No differences in intensity of Alizarin staining, elements stained, or cartilage architecture were noted in the rspo3−/− fish. Scale bar: 100 μm. (B) Lateral view of 14 dpf showing increased osteoclast activity (red stain in the black dotted box) in rspo3 mutant as compared to wild-type. At 21 dpf, more areas of osteoclast activity in the dentary, hyomandibular, pharyngeal teeth and jaws (solid arrow) were observed in rspo3−/− compared to wild-type. Quantification of total area of red staining. P < 0.05. N = 5. Scale bar: 100 μm.
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