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Fig. 4. Defects of calvarial osteoblast differentiation caused by the Axin2
mutation. Primary calvarial osteoblast precursors isolated from the
Axin2+/+ and Axin2–/–
littermates were cultured in differentiation media for up to 9 days. Liquid
(A) and histochemical (B) assays for alkaline phosphatase were performed at
different time points of osteoblast (OB) differentiation as indicated. The
enzyme activity is expressed as micrograms of p-nitrophenol (Pi)
released per microgram of protein per minute. A diagram summarizes neural
crest contribution (blue) to the skeletal elements and sutures of the mouse
skull vault [diagram modified from Jiang et al.
(Jiang et al., 2002)] (C). The
neural crest or mesoderm-derived osteoblasts were isolated from nasal/frontal
(highlighted by a red line) or parietal (highlighted by a green line) bones,
respectively. c, coronal suture; F, frontal bone; IP, interparietal bone; JL,
jugum limitans; l, lambdoid suture; m, metopic suture; N, nasal bone; nc,
nasal cartilage; P, parietal bone; s, sagittal suture. (D) The neural crest or
mesoderm-derived primary osteoblasts of Axin2+/+ and
Axin2–/– were cultured in differentiation
media for up to 12 days. Liquid assays for alkaline phosphatase were performed
at different time points as indicated. A and D are representative of three
independent experiments. Scale bar: 200 µm in B.
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