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Fig. 3. Alteration of bone formation and resorption in Bmpr1a cKO
mice. (A) QRT-PCR for bone formation markers (Runx2, Sp7, Ibsp,
Akp2 and Bglap2 using E16.5 and E18.5 calvariae). Values are
expressed relative to wild type at E16.5. (B) QRT-PCR for bone
resorption markers expressed by osteoclasts (Mmp9, Ctsk and
Trap), and Rankl and Opg expressed by osteoblasts
using E18.5 and E19.5 calvariae. Values are expressed relative to wild type at
E18.5. (C) Relative ratio of Rankl to Opg expression
levels calculated from Fig. 3B. Values are expressed relative to wild type at
E18.5. (D) Evaluation of osteoclast activity by TRAP staining using
E18.5 calvariae. The positive cells localized randomly in cKO calvariae
compared with wild type (left two panels, red arrows). The percent of
TRAP-positive cells per total cells in bone area detected by DAPI was
significantly reduced in cKO calvariae (wild type, 13.9%; cKO, 6.4%, right
panel). Scale bars: 50 µm. (E) QRT-PCR for BMP type I receptors
(Bmpr1b, Acvr1), type II receptors (Bmpr2, Acvr2a and
Acvr2b) and potential ligands for these receptors (Bmp2, Bmp4,
Bmp6 and Bmp7) using E18.5 calvariae. Values in A-E represent
mean±s.d. from a minimum of three independent experiments using
wild-type and cKO bones. Student's t-test;
*P<0.05.
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