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Fig. 5 2D assessment of the LP-driven increase in bone matrix volume that subsequently mineralizes with sufficient-P intake. Sagittal (A and C) and cross (B) non-demineralized histological sections of the vertebral column stained with Von Kossa/Van Gieson. Compared to controls, LP–LP, LP–RP, and LP–HP animals show an increase in bone volume in the middle region of the vertebral centrum (white arrowheads in A, black arrowheads in B), at the arches (white arrowheads in B), and at the vertebral body endplates (pink arrowheads, C). In LP–LP animals, the amount of non-mineralized bone matrix (pink staining) is larger compared to that in LP–RP and LP–HP animals, where the bone structures are mineralized (black staining). In all zebrafish with LP dietary history, the enlarged bone fills the space occupied by fat in RP–RP (white asterisks). Nt: neural tube. (D and E) Histology-based measurements. (D) The total bone area (mineralized and non-mineralized) at the vertebral body endplates, normalized to vertebral body length, is significantly increased by 1.5-fold in LP–LP, LP–RP, and LP–HP zebrafish compared to controls (RP–RP). (E) At the vertebral body endplates, the mineralized bone area divided by the total bone area, used as indicator of mineralization, shows that bone mineralization depends on dietary P, and non-mineralized bone generated during the LP period can resume mineralization to an extent similar to controls when sufficient-P is provided with the diet. Values are reported with Box and Whisker plots: the midline in each box is indicative of the median, whereas min and max values are shown with whiskers. Individual data points and P values are shown.