Disruption of Dhcr7 and Insig1/2 in cholesterol metabolism causes defects in bone formation and homeostasis through primary cilium formation

摘要

Loss of accelerates osteogenesis. MicroCT images of the calvaria—top view (upper panels) and side view (lower panels)—of newborn wild-type (WT) control and knockout (KO) mice. Red arrows indicate early closure of the metopic, coronal, and sagittal sutures. Yellow dotted lines in the lower panels indicate the tips of premaxillae. Skeletal staining of skulls from newborn WT control and KO mice. Yellow boxed areas are enlarged in the lower images. Black arrows in KO images indicate overlapping of frontal and parietal bones at the coronal suture and left and right parietal bones at the sagittal suture. Hematoxylin and Eosin (H&E) staining of the sagittal sutures of newborn WT and KO mice. Arrows indicate the osteogenic front. Scale bar, 50 µm. von Kossa staining of the sagittal sutures of newborn WT and KO mice. Arrows indicate the osteogenic front. Scale bar, 100 µm. Immunoblotting for type I collagen (COL1) in P0 calvaria of WT and KO mice. GADPH was used as loading control. Quantitative RT-PCR of the indicated osteogenic genes at E14.5 (left) and E16.5 (right) in WT (blue bars) and KO (red bars) mice.  = 6 per genotype per stage. **  P g Immunohistochemistry analysis for RUNX2, COL1A1 and SP7 (Osterix) in newborn WT and KO mice. Nuclei were counterstained with 0.04% methylene blue. Scale bar, 50 µm. Alkaline phosphatase (left) and Alizarin Red (right) staining of osteoblasts isolated from newborn WT and KO calvaria after induction of osteogenic differentiation at Day 0, 7, and 14.