Article Title: Altered bone growth dynamics prefigure craniosynostosis in a zebrafish model of Saethre-Chotzen syndrome
Figure Lengend Snippet: Altered bone growth dynamics in Tcf12 +/- ; Twist1 +/- mice. ( A ) Lateral views of E13.5 mouse heads show Alkaline phosphatase staining of developing frontal and parietal bones. Dotted lines indicate the bone fronts that will form the coronal suture. Compared to wild types ( n = 5), the fronts were accelerated in all Tcf12 +/- ; Twist1 +/- mutants ( n = 12). Scale bar, 1 mm. ( B ) Dorsal views of skull bones stained with Alizarin Red at birth (P0). Compared to wild types ( n = 5), the fronts were closer together in all Tcf12 +/- ; Twist1 +/- mutants ( n = 11). Scale bar, 1 mm. ( C ) Sections of E14.5 coronal sutures and E16.5 sagittal sutures stained for BrdU (magenta), Sp7 protein (green), and DAPI (blue, nuclei). Boxed regions are magnified in lower panels, with yellow dotted lines indicating the regions of interest (ROI) used for quantification. fb, frontal bone; pb, parietal bone. Scale bar, 100 µm. ( D–F ) Quantification of Sp7 + osteoblasts per ROI ( D ), BrdU + Sp7 - bone front cells per ROI ( E ), and the percentage of Sp7 + osteoblasts that are BrdU + in the ROI ( F ). Cell counts were performed at the developing coronal sutures (cs, four wild types, three mutants) and sagittal sutures (ss, six wild types, six mutants). p values were determined by Student’s t-tests; error bars represent standard error of the mean.
Article Snippet: Immunohistochemistry was performed using rat anti-BrdU (MCA2060 GA, Bio-Rad), rabbit anti-Osx/Sp7 (sc-22536-r, Santa Cruz), goat anti-Grem1 (PA5-47973, Thermo Fisher), or goat anti-Gli1 (AF3455, R and D Systems) diluted in 1% BSA/PBS and incubated overnight at 4°C.
Techniques: Mouse Assay, Staining