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. 2011 Apr 25;6(4):e19313. doi: 10.1371/journal.pone.0019313

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Li et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Micro-CT analysis of total body (left panels) and femur of 4-week-old Lmna ^−/− mice and WT littermates. Images representative of two-and three- dimensional reconstructions, obtained with a 0.9° rotation between frames on a Skyscan® 1072 instrument, are shown for WT mice (A, upper panel) and Lmna ^−/− (A, lower panel) mice. Whole-body CT analysis of Lmna ^−/− mice and WT littermate controls shows dramatically lower bone accumulation in Lmna ^−/− mice with remarkable skull defects observed in the mutant mice. The right upper panels are representative of longitudinal sections. The lower right panels show the metaphysis (area just below the growth plate) (M) and distal diaphysis (cortical structure) (D). Lmna ^−/− mice exhibited profound thinning of cortical bone, a reduction in platelike structures and a lack of trabecular connectivity. These changes correlated with von Kossa staining (B). Quantitation of bone parameters (C) further exemplified a decrease in bone volume vs. total volume (BV/TV), trabecular number (Tb.N), and cortical thickness (Ct.Th) with a concomitant increase in trabecular separation (Tb.Sp) in the mutant femora compared with the WT littermate controls. Results are expressed as the mean ± SD of eight independent analyses per group. *P<0.001, significantly different from null mice. (D) Tetracycline-labeled section of the distal femur. The distance between two layers of tetracycline labels (arrows) visualized by epifluorescence represents bone formation that occurred during the 5-d period between tetracycline injections. Mutant mice showed a significant decrease in all parameters of bone formation including mineralized surface/bone surface (MS/BS), mineral apposition rate (MAR) and bone formation rate/bone surface (BFR/BS) as compared with their WT littermates (*P<0.001). Photomicrographs were obtained on the Bioquant analysis system using a ×40 objective.