Paradoxical effects of partial leptin deficiency on bone in growing female mice

Abstract

Morbidly obese, leptin-deficient ob/ob mice display low bone mass, mild osteoclast-rich osteopetrosis, and increased bone marrow adiposity. While partial leptin deficiency results in increased weight, the skeletal manifestations of partial leptin deficiency are less well defined. We therefore analyzed femora and lumbar vertebrae in growing (7-week-old) female C57BL/6 wildtype (WT) mice, partial leptin-deficient ob/+ mice, and leptin-deficient ob/ob mice. The bones were evaluated by dual energy absorptiometry, microcomputed tomography and histomorphometry. As expected, ob/+ mice were heavier, had more white adipose tissue, and lower serum leptin than WT mice, but were lighter and had less white adipose tissue than ob/ob mice. With a few exceptions, cancellous bone architecture, cell (osteoblast, osteoclast, and adipocyte), and dynamic measurements did not differ between WT and ob/+ mice. In contrast, compared to WT and ob/+ mice, ob/ob mice had lower cancellous bone volume fraction, and higher bone marrow adiposity in the femur metaphysis, and higher cancellous bone volume fraction in lumbar vertebra. Paradoxically, ob/+ mice had greater femoral bone volume than either WT or ob/ob mice. There was a positive correlation between body weight and femur volume in all three genotypes. However, the positive effect of weight on bone occurred with lower body weight in leptin-producing mice. The paradoxical differences in bone size among WT, ob/+, and ob/ob mice may be explained if leptin, in addition to stimulating bone growth and cancellous bone turnover, acts to lower the set-point at which increased body weight leads to a commensurate increase in bone size.

Keywords: histomorphometry; microcomputed tomography; ob/+ mice; ob/ob mice; obesity.

Figure 1

Effects of genotype on body weight (A), abdominal white adipose tissue weight (B), food consumption (C), uterine weight (D), serum leptin (E), and serum CTx (F) in 7-week-old female WT, ob/+, and ob/ob mice. Data are mean ± SE. ^aDifferent from WT, P < 0.05; ^bDifferent from ob/+, P < 0.05.

Figure 2

Effects of genotype on femur bone mineral content (A), bone area (B), bone mineral density (C), length (D), total bone volume (E), total bone volume to length ratio (F), mid-diaphysis cross-sectional area (G), cortical area (H), marrow area (I), cortical thickness (J), and polar moment of inertia (K) in 7-week-old female WT, ob/+, and ob/ob mice. Data are mean ± SE. ^aDifferent from WT, P < 0.05; ^a*Different from WT, P < 0.1; ^bDifferent from ob/+, P < 0.05.

Figure 3

Correlation between body weight and total femur bone volume (A) and body weight and total vertebra bone volume (B) in 7-week-old WT, ob/+, and ob/ob mice.

Figure 4

Effects of genotype on distal femur longitudinal growth rate (A), growth plate thickness (B), hypertrophic zone thickness (C), and proliferative zone thickness (D) in 7-week-old female WT, ob/+, and ob/ob mice. Data are mean ± SE. ^aDifferent from WT, P < 0.05; ^bDifferent from ob/+, P < 0.05.

Figure 5

Effects of genotype on distal femur metaphysis mineralizing perimeter (A), mineral apposition rate (B), bone formation rate (C), osteoblast perimeter (D), osteoclast perimeter (E), bone marrow adiposity (F), and bone marrow adipocyte density (G) in 7-week-old female WT, ob/+, and ob/ob mice. Representative histological sections illustrating differences in bone marrow adiposity and fluorochrome labeling among the 3 genotypes are shown in H-J. Data are mean ± SE. ^aDifferent from WT, P < 0.05; ^bDifferent from ob/+, P < 0.05. Scale bar, 200 μm.

Figure 6

Effects of genotype on vertebral mineralizing perimeter (A), mineral apposition rate (B), bone formation rate (C), osteoblast perimeter (D) and osteoclast perimeter (E) in 7-week-old WT, ob/+, and ob/ob mice. Data are mean ± SE. ^aDifferent from WT, P < 0.05; ^bDifferent from ob/+, P < 0.05; ^b*Different from ob/+, P < 0.1.