6C; Wt = 15.9 ± 3.9 N; Mecp2stop/y = 12.6 ± 2.4 N; Mecp2stop/y, CreER = 13.4 ± 2.2 N, n = 5 per genotype, p > 0.05, ANOVA with Tukey’s post hoc test). Similar findings were obtained in the female groups ( Fig. 7). Picrosirius red staining of the femur was used to assess Y-27632 collagen content (Fig. 8A) as described previously [39].

Mecp2stop/y mice showed a significant decrease (− 24%) in collagen content compared to Wt mice ( Fig. 8B; Wt = 65.1 ± 8.6%; Mecp2stop/y = 48.8 ± 9.1%; Mecp2stop/y, CreER = 55.63 ± 11.4%; n = 10 per genotype, p < 0.01, one way ANOVA with Tukey's post hoc test). TRAP staining was conducted to assess resorption activity (osteoclast number per bone surface), but showed no difference between genotypes (Wt = 12.61 ± 8.51; Mecp2stop/y = 17.48 ± 6.13; Mecp2stop/y, CreER = 18.90 ± 4.61; n = 5 per genotype, p > 0.05, one way ANOVA with Tukey’s post hoc test). Qualitative analysis using scanning electron microscopy (SEM) of the

distal femur (n = 5 per genotype) revealed porous structure in cortical ABT199 bone (3 of 5 mice) as well as alterations in the architecture of trabecular bone in Mecp2stop/y mice ( Fig. 9A–B). The central metaphyseal region in Mecp2stop/y mice showed a sparse trabecular mass consisting of short, thin trabecular rod and plate structures. In contrast, a more robust trabecular structure, with a network of shorter and thicker rods and plates was found in wild-type control tissue ( isothipendyl Fig. 9Ai–ii). The porosity and altered trabecular structure was less evident in rescued Mecp2stop/y, CreER mice ( Fig. 9C). These features were investigated further and a quantitative manner using μCT (below). In contrast to the male mice, we did not observe overt tissues differences in heterozygous Mecp2stop/+ mice. Three dimensional μCT analysis was performed to obtain a quantitative measure of trabecular architecture in wild-type, Mecp2stop/y and Mecp2stop/y, CreER mouse lumbar 5 (L5) vertebrae (

Fig. 10A). A significant reduction of L5 trabecular thickness (~ 30%) was observed in Mecp2stop/y mouse tissues compared to the wild-type control. Interestingly Mecp2stop/y, CreER mouse L5 μCT results, showed a significant increase (+ 80%, p < 0.01) in trabecular rod and plates thickness compared to Mecp2stop/y mice ( Fig. 10B–E; Wt = 0.073 ± 0.01 mm; Mecp2stop/y = 0.051 ± 0.02 mm; Mecp2stop/y, CreER = 0.09 ± 0.02 mm; n = 7 per genotype; p < 0.01, ANOVA with Tukey’s post hoc test). No significant differences were observed in trabecular separation, trabecular bone volume, trabecular porosity, bone mineral density (BMD), degree of anisotropy (DA) and structure model index (SMI) between genotypes ( Table 2). μCT analysis of tibia showed a significant difference in cortical bone thickness, outer perimeter length, inner perimeter length, marrow area, total area and bone volume in Mecp2stop/y mouse compared to wild-type controls (p < 0.05, n = 7 per genotype, ANOVA with Tukey’s post hoc test).