Fig. 8

Impairment of vertebral development persists following recovery from acute hyperglycemia. (A and B) Lateral view of C22 and Tx22 zebrafish stained with alizarin red solution; robust differences are observed in terms of mineralization of centra, development of neural arches (na; extending dorsally from the centra) and hemal arches (ha; extending ventrally from the centra), and mineralization of the anal fin rays (A and B, arrows) between C22 and Tx22 zebrafish. Tx22 fish continued to exhibit statistically significant reductions in the area of mineralized centra (V3-5) and increased intervertebral domain (IVD) area compared to C22 fish, and these differences were significant (“All” bars in C and D, respectively, ****P < .0001). (C) This difference in centra area persisted when Tx22 and C22 fish were size matched by total body length (TL): 4 mm, ****P < .0001; 5 mm, ****P < .0001, 6 mm, ***P = .0003. No analyses were conducted at 7 and 8 mm TL as there were no Tx22 of this size. (D) Likewise, the difference in IVD area persisted when Tx22 and C22 were size matched by TL: 4 mm, **P = .0029; 5 mm, ****P < .0001; 6 mm, ***P = .0004. (E) Tx22 fish were significantly less likely to have neural arches present anywhere along their axial skeleton compared to C22 fish, ****P < .0001. (F) A significant absence of hemal arches in Tx22 fish was also observed, ****P < .0001. (G) Mineralization of skeletal structures within the caudal fin were also observed in C22 fish. (H) Mineralization of the caudal vertebrae and fin rays were absent in Tx22 fish. All values are means ± SEM (see Table 1). Scale bars = 200 μm.