Impaired growth plate function in bmp-6 null mice

Abstract

Bone morphogenetic protein 6 (BMP-6) is expressed by different skeletal cells including osteoblasts and growth plate chondrocytes, suggesting roles in bone formation and growth regulation. To address these possibilities, we examined whether cancellous and cortical bone parameters, or indices of growth plate function, are altered in bmp-6 null mice as assessed under basal conditions, and following stimulation of bone formation and suppression of growth by estrogen treatment. Ten-week-old female littermate bmp-6 null and wild-type (WT) mice were administered vehicle or E 2 4, 40, 400 or 4000 μg/kg/day by daily sc injection for 28 days (6–8 per group). Tibias were removed, and detailed histomorphometric analysis of the proximal metaphysis and growth plates, and tibial diaphysis were performed on longitudinal and transverse sections respectively. Long bone area as measured by DXA was reduced in vehicle-treated bmp-6 null mice compared with WT littermate controls. In addition, vehicle-treated bmp-6 null mice had a reduced cross-sectional area at the tibial mid-diaphysis as assessed by histomorphometry, whereas cancellous bone indices were unaffected. Histomorphometry of the proximal tibial metaphysis demonstrated a defect in bone formation immediately adjacent to the growth plate in bmp-6 null mice compared to WT mice following E 2 treatment. E 2 administration was also associated with a dose-responsive decrease in longitudinal growth rate, and proliferative and hypertrophic zone parameters of the growth plate ( p < 0.0001). Significantly greater reductions following E 2 treatment were observed in longitudinal growth rate ( p < 0.01), proliferating and hypertorphic zone widths ( p < 0.001), and proliferating ( p < 0.0002) and hypertrophic ( p < 0.002) cells per column of bmp-6 null mice compared to WT mice. Our observation that long bones are reduced in size compared to wild-type mice primarily through a decrease in cortical cross-sectional area, whilst cancellous bone mass is unaltered, suggests a non-redundant role for BMP-6 in periosteal but not trabecular bone formation. Moreover, growth plate function was reduced in bmp-6 null mice receiving estrogen, leading to an impaired cancellous bone response to estrogen at the highest dose, suggesting that BMP-6 also plays a physiological role in maintaining growth plate function.