Notch1 and Notch2 expression in osteoblast precursors regulates femoral microarchitecture

Notch receptors regulate cell differentiation and function. Notch1 and Notch2 inactivation in osteoblasts and osteocytes increases cancellous bone volume, but the function of Notch signaling in osteoblast precursors is unknown. To inactivate Notch signaling in immature osteoblastic cells, mice homozygous for conditional Notch1 and Notch2 alleles (Notch1(loxP/loxP);Notch2(loxP/loxP)) were crossed with mice where the osterix (Osx) promoter, regulated by a Tet-Off cassette, governs Cre expression (Osx-Cre). Notch1(loxP/loxP);Notch2(loxP/loxP) control and Osx-Cre(+/-);Notch1(Δ/Δ);Notch2(Δ/Δ) experimental littermate cohorts were obtained. To prevent the effects of embryonic Osx-Cre expression, doxycycline was administered to pregnant dams, but not to newborns. Recombination of conditional alleles was documented in calvarial DNA extracts from 1month old mice. Notch1 and Notch2 inactivation did not affect femoral microarchitecture at 1month of age. Cancellous bone volume was higher and structure model index was lower in 3 and 6 month old Osx-Cre(+/-);Notch1(Δ/Δ);Notch2(Δ/Δ) mice than in control littermates and the effect was more pronounced in female mice. One month old Osx-Cre(+/-);Notch1(Δ/Δ);Notch2(Δ/Δ) male mice transiently exhibited an increase in osteoblast number and a modest suppression in bone resorption. Osx-Cre(+/-);Notch1(Δ/Δ);Notch2(Δ/Δ) female mice displayed a tendency toward increased bone formation at 3months of age, although bone remodeling was suppressed in 6month old Osx-Cre(+/-);Notch1(Δ/Δ);Notch2(Δ/Δ) female mice. Notch1 and Notch2 inactivation increased porosity and reduced thickness of cortical bone. These effects were modest and more evident in 3 and 6 month old female than in male mice of the same age. In conclusion, Notch1 and Notch2 expression in osteoblast precursors regulates cancellous bone volume and microarchitecture.