Segments of neonatal mouse bone were prepared by using a modification of the method described by Mohammad et al. [ 30].
The developmental expression pattern of OPN has not been previously investigated and, therefore, our result demonstrating temporally limited overexpression in mouse bone is interesting (Fig. 3).
This method enables us to analyze precisely comparable sites of the loaded and contra-lateral control tibiae because the effects of loading are site-specific and the mouse bone is small.
On the other hand, the reported crystal size from fish, bovine, chicken, and mouse bones was similar, about 40 nm in length and 30 nm in width, with different distribution in each animal studied [ 8].
The mouse bones were carefully oriented for longitudinal sectioning to include observing the epiphyseal plate as well as the distal region of the bone in the same section (Fig. 3A, top panel).
Gross abnormalities of KO mouse bones were not observed as shown in the X-ray of bone from both FL and KO mice (Figure 7), suggesting that even OGR1 might even have a role in osteoclastogenesis, the overall effects are not apparent and that a redundant role of other genes may be involved in vivo.
For analysis of bone marrow endothelium, mouse bones were harvested 4 d post-irradiation, crushed and then stained with a panel of haematopoietic/endothelial markers exactly as described previously by Hooper et al (2009).
In addition, our preliminary data using adult mice bones are also presented in this table, and demonstrates positional identity marker expression, at least, appears to be conserved between these two species).
BMD and bone mineral content (BMC) of mouse femoral bone were determined with a PIXImus mouse densitometer (GE Lunar, Waukesha, WI, USA).
Intravital microscopy of mouse bone marrow was performed using a protocol modified from a previous report [38].
