Table 1.
List of genes and primers used to analyze gene expression by quantitative real time PCR.
Table 2.
Comparison of wild type (WT) and Cox2 KO mice.
Table 3.
Body weight, serum calcium (Ca) and serum PGE2 in wild type (WT) and Cox2 KO mice infused with vehicle or PTH for 12 days.
Fig 1.
Femoral BMD and serum markers of bone turnover in mice infused for 12 days.
A. BMD was measured in vivo at beginning and end of infusion with vehicle (VEH) or PTH (40 μg/kg/d) in WT and Cox2 KO mice and the percent change calculated relative to the beginning BMD for each mouse. B, C. Serum parameters for bone formation (P1NP) and bone resorption (CTX) were measured at the time of euthanasia. Bars are means ± SEM for 7 WT and 7 KO mice treated with vehicle and 8 WT and 8 KO treated with PTH. aSignificant effect of PTH, p<0.01. bSignificant effect of genotype, p<0.01.
Fig 2.
Morphometry of femoral bone and L3 vertebral bone in mice infused for 12 days.
A. Representative μCT longitudinal images (top panel) and morphometric analyses (bottom panel) of trabecular bone in the metaphyseal region of distal femur. Bars are means ± SEM for 7 WT and 7 KO mice treated with vehicle and 8 WT and 8 KO mice treated with PTH. B. Representative μCT cross-sectional images (top panel) and morphometric analyses (bottom panel) of trabecular bone in the L3 vertebrae. Bars are means ± SEM for 7 WT and 7 KO mice treated with vehicle and 8 WT and 8 KO mice treated with PTH. C. Representative μCT cross-sectional images of midshaft of femurs. aSignificant effect of PTH, p<0.01; bp<0.05. cSignificant effect of genotype, p<0.01; dp<0.05.
Table 4.
Cortical morphometry by μCT in the midshaft femoral region of WT and Cox2 KO mice after infusion with vehicle or PTH for 12 days.
Fig 3.
Static and dynamic histomorphometric analysis of distal femurs in mice infused for 12 days.
A. Representative microscopic images of distal femur. (1) Hematoxylin staining at 20x magnification. (2) Tartrate resistant acid phosphatase (TRAP) staining and counter staining with hematoxylin at 400x magnification in PTH-infused WT and KO mice. (3) Double labeling with calcein (green) and demeclocycline (orange/brown) of trabeculae at 400x magnification in PTH-infused WT and KO mice. B. Histomorphometric analysis of distal femurs. Bars are means ± SEM for 7 WT and 7 KO mice treated with vehicle and 8 WT and 8 KO mice treated with PTH. aSignificant effect of PTH, p<0.01; bp<0.05. cSignificant effect of genotype, p<0.01.
Fig 4.
Tibial gene expression in mice infused with vehicle or PTH for 12 days.
At the end of infusion, both tibiae from each mouse, minus ends, were combined for RNA extraction. mRNA expression was measured by qPCR, as described in Methods. Bars are means ± SEM for 7 WT and 7 KO mice infused with vehicle and 8 WT and 8 KO infused with PTH. aSignificant effect of PTH, p<0.01;bp<0.05. cSignificant effect of genotype, p<0.01.
Table 5.
Body weight and serum calcium (Ca) in 3-mo old wild type (WT) and Cox2 KO mice infused with vehicle or PTH for 21 days.
Fig 5.
Femoral BMD and serum markers of bone turnover in mice infused for 21 days.
A. BMD was measured in vivo at beginning and end of infusion with vehicle or PTH in WT and Cox2 KO mice and the percent change calculated relative to the beginning BMD for each mouse. B, C. Serum parameters for bone formation (osteocalcin) and bone resorption (CTX) were measured at the end of study. Bars are means ± SEM for 6 WT and 6 KO mice infused with vehicle and 7 WT and 7 KO infused with PTH. aSignificant effect of PTH, p<0.01; bp<0.05. cSignificant effect of genotype, p<0.01.
Fig 6.
Morphometry of femoral trabecular bone and L3 vertebrae in mice infused for 21 days.
A. Representative μCT images (top panel) and morphometric analyses (bottom panel) of trabecular bone in the metaphyseal region of distal femur. B. Representative μCT images (top panel) and morphometric analyses (bottom panel) of trabecular bone in L3 vertebrae. Bars are means ± SEM for 6 WT and 6 KO mice infused with vehicle and 7 WT and 7 KO mice infused with PTH. aSignificant effect of PTH, p<0.01; bp<0.05. cSignificant effect of genotype, p<0.01. C. Representative μCT cross-sectional images of midshaft of femurs.
Table 6.
Cortical morphometry by μCT in the midshaft femoral region of WT and Cox2 KO mice after infusion with vehicle or PTH for 21 days.
Fig 7.
Tibial gene expression in mice infused with vehicle or PTH for 21 days.
At the end of infusion, both tibiae were combined for RNA extraction and mRNA expression was measured by qPCR Bars are means ± SEM for 6 WT and 6 KO mice infused with vehicle and 7 WT and 7 KO infused with PTH. aSignificant effect of PTH, p<0.01; bp<0.05. cSignificant effect of genotype, p<0.01; dp<0.05.