Static histomorphometric parameters and cellular activities in femur bones from 3, 10 and 16 week old BSP+/+ (+/+) and BSP−/− (−/−) mice.

<p>Data are Mean±SEM of N = 6 to 10 mice; *:p<0.05, **:p<0.001 vs BSP+/+, Mann-Whitney U Test.</p

Growth plate and osteoblast marker protein expression and blood levels in newborn and adult BSP+/+ and BSP−/− mice.

<p>QRT-PCR was performed 6 days after birth on collected whole femurs and tibias including the growth plates, ground and extracted in Tri-reagent. Messenger RNA expression of growth plate regulators IHH, PTHrP and IGF-1 (A), markers of bone formation Runx2, Osx and Ocn (B) and SIBLING proteins DMP1, MEPE and Opn (C) were assessed in samples from BSP+/+ (N = 5) and BSP−/− (N = 9) mice. Expression levels are normalized on the housekeeping gene GAPDH, and on Runx2 levels for Opn (C). ELISA assay of Opn (D) in the serum of 6 days (N = 5 pools), 35 days and 12 month old (N = 5 male) BSP+/+ and BSP−/− mice. Data are Mean±SEM; *: p<0.05, **: p<0.01, ***: p<0.001 vs BSP+/+, Mann-Whitney U test.</p

Growth plate kinetics of newborn, 3 week old, 10 and 16 week old BSP+/+ and BSP−/− mice.

<p>Data are Mean±SEM of N = 7 to 8 newborn and 4 to 6 adult mice; *:p<0.05, **:p<0.001 vs BSP+/+ Mann-Whitney U Test. Micrographs of the growth plate in the distal femur of BSP+/+ and BSP−/− newborn and 3, 10 and 16 week old mice.</p

Morphology and morphometry of the long bones in newborn BSP+/+ and BSP−/− mice.

<p>(A) 3D µCT reconstruction and morphometry of newborn whole femurs from BSP+/+ (+/+) and −/− newborn mice. (B) Histomorphometry of trabecular bone in newborn femurs from +/+ and −/− mice. Data are Mean±SEM of N = 4 to 10 mice; *:p<0.05, **:p<0.01 Vs +/+, Mann-Whitney U Test. (C) Micrographs of Goldner's trichrome and Von Kossa staining of BSP+/+ and BSP−/− newborn femur sagittal sections. Yellow polygon: ROI for histomorphometry of the trabecular bone. Bar  = 400 µm.</p

Newborn morphology and growth of BSP+/+ and BSP−/− mice.

<p>(A) general aspect of BSP+/+ (+/+) and BSP −/− (−/−) newborn mice. (B) Kinetics of weight gain during the growth of BSP+/+ and BSP−/− mice. M, males; F, females. Data are Mean±SEM of N = 4 to 31 mice. Note the log scale.</p

PCR primer sequences in 5′-3′ direction.

<p>The meaning of the abbreviations is explained in the text.</p

Impact of the lack of BSP on mother's behavior.

<p>(A) Pictures of BSP+/+ and BSP−/− mothers with pups; note the well formed "nest" in the wild type mouse cage as opposed to the untouched and spread out pieces of soft paper in the mutant mouse cage (arrows). Note: the 129 sv/CD1 background is outbred, leading to hair color variability. (B) Body length (left) and weight (right) increase in the 30 first days of growing male and female BSP+/− mice from BSP+/+ (○, N = 7 pups) and BSP−/− mothers (•, N = 6 pups). (C) Femur and tibia length of 40 days old BSP+/− mice from either BSP+/+ or BSP−/− mothers. (D) Femur and tibia length of 40 days old BSP+/− (N = 6) and BSP−/− mice (N = 6) from BSP−/− mothers crossed with BSP+/− males. (E) Body weight and femur length of 40 days old BSP+/+ (N = 7), BSP−/− (N = 11) and BSP+/− mice (N = 22) from BSP+/− parents. Data are Mean±SEM; *: p<0.05 vs BSP+/+ and/or BSP+/−, Mann-Whitney U Test or (E) Kuskall-Wallis with post-test.</p

Impact of the absence of BSP on postnatal development of the skeleton.

<p>The table compares to wild type (+/+), at different ages of developing mutant mouse long bones, the whole growth plate (GP), hypertrophic (HZ) and proliferating zone (PZ) thickness, cortical bone thickness, trabecular bone volume (BV/TV), osteoclast surface/number and osteoid thickness.</p

Skeletal development and mineralization of newborn BSP+/+ and BSP−/− mice.

<p>(A) Skeletal preparation (alcian blue/alizarin red staining) of BSP+/+ (+/+) and BSP −/− (−/−) E19 mice. (B) 3D µCT reconstruction of whole newborn +/+ and −/− mice, showing undermineralized fingers (arrows and black-bordered detailed pictures), and (C) high magnification of the heads showing the widened sutures (arrows).</p