Figure 1.

Maternal B₁₂ supplementation in the wild-type mice during gestation increases bone mass accrual of the offspring during postnatal growth. A: schematic representation of the experimental protocol used. Body weight analysis (B), tibia length (C), tibia weight (D), representative images of Von Kossa/Van Gieson-stained vertebral sections (area used to measure BV/TV% is shown) (E), bone volume over total volume % (BV/TV%) in L4 vertebral sections of the offspring (F), osteoblast number per trabecular area (G), bone formation rate (H), and mineral apposition rate (I), osteoclast surface/bone surface % (J), serum osteocalcin levels (K), serum deoxypyridinoline (dpd; L) levels in offspring from mothers that received either vehicle (0) or B₁₂ daily through subcutaneous injections at the dose of 20 or 200 μg/day during gestation from embryonic day 0.5 to birth. Micro-CT analysis of tibia showing BV/TV% (M), trabecular numbers (N), trabecular thickness (O), and cortical thickness (P). Gene expression analysis of markers of osteoblast differentiation (Q) and serum taurine levels in offspring (R). S: histomorphometric analysis of L4 vertebra of offspring at 1 yr of age from mothers treated with either 0 or 200 μg/day dosage of B₁₂ during gestation. Histological and histomorphometric analysis of L4 vertebra in mothers that received either vehicle (0) or B₁₂ daily through subcutaneous injections at the dose of 20 or 200 μg/day during pregnancy: representative images of Von Kossa/Van Gieson-stained vertebral sections (area used to measure BV/TV% is shown) (T), BV/TV% (U), Osb.N./T.ar. (V), and Oc.S./BS% (W) is shown. Means ± SE is shown. n = 8–10 (A–R), n = 5 (S), n = 8 (T–W) female mice each group. *P < 0.05. #P < 0.01.