Cortical and trabecular morphology is altered in the limb bones of mice artificially selected for faster skeletal growth.

Bone strength is influenced by mineral density and macro- and microstructure. Research into factors that contribute to bone morphology and strength has focused on genetic, environmental and morphological factors (e.g., body mass index), but little is known regarding the impact of rates of skeletal elongation on adult skeletal morphology and strength. Using micro-CT, we examined the impact of rates of skeletal elongation on bone cortical and trabecular morphology, and on rates of estrogen-dependent bone loss in the tibia in CD-1 mice, and in mice with accelerated skeletal growth (Longshanks). Groups of adult mice (n = 7/group) were subjected to ovariectomy or sham surgeries, scanned for 6 weeks, and indices of bone morphology were collected. Results show that Longshanks mice had significantly less trabecular bone at skeletal maturity, characterized by fewer, thinner trabeculae, and furthermore lost trabecular bone more slowly in response to ovariectomy. Artificial selection for rapid skeletal growth relative to somatic growth thus had a significant impact on trabecular bone morphology in Longshanks. Our data do not unequivocally demonstrate a causal relationship between rapid bone growth and reduced trabecular bone quality, but suggest that rapid linear bone growth may influence the risk of cancellous bone fragility.

Gait changes in a line of mice artificially selected for longer limbs.

In legged terrestrial locomotion, the duration of stance phase, i.e., when limbs are in contact with the substrate, is positively correlated with limb length, and negatively correlated with the metabolic cost of transport. These relationships are well documented at the interspecific level, across a broad range of body sizes and travel speeds. However, such relationships are harder to evaluate within species (i.e., where natural selection operates), largely for practical reasons, including low population variance in limb length, and the presence of confounding factors such as body mass, or training. Here, we compared spatiotemporal kinematics of gait in Longshanks, a long-legged mouse line created through artificial selection, and in random-bred, mass-matched Control mice raised under identical conditions. We used a gait treadmill to test the hypothesis that Longshanks have longer stance phases and stride lengths, and decreased stride frequencies in both fore- and hind limbs, compared with Controls. Our results indicate that gait differs significantly between the two groups. Specifically, and as hypothesized, stance duration and stride length are 8-10% greater in Longshanks, while stride frequency is 8% lower than in Controls. However, there was no difference in the touch-down timing and sequence of the paws between the two lines. Taken together, these data suggest that, for a given speed, Longshanks mice take significantly fewer, longer steps to cover the same distance or running time compared to Controls, with important implications for other measures of variation among individuals in whole-organism performance, such as the metabolic cost of transport.

Changes in shape and cross-sectional geometry in the tibia of mice selectively bred for increases in relative bone length.

Limb bone size and shape in terrestrial mammals scales predictably with body mass. Weight-bearing limb bones in these species have geometries that enable them to withstand deformations due to loading, both within and between species. Departures from the expected scaling of bone size and shape to body mass occur in mammals that have become specialized for different types of locomotion. For example, mammals adapted for frequent running and jumping behaviors have hind limb bones that are long in relation to body mass, but with narrower cross-sections than predicted for their length. The Longshanks mouse was recently established, a selectively bred line of mice with ~12-13% longer tibiae relative to body mass. This increased limb length resembles superficially the derived limb proportions of rodents adapted for hopping and jumping. Here, 3D geometric morphometrics and analyses of bone cross-sectional geometry were combined to determine whether selection for increased relative tibia length in Longshanks mice has altered the scaling relationship of size and shape, and/or bone robusticity, relative to the tibiae of random-bred control mice from the same genetic background. The results suggest that the Longshanks tibia is not a geometrically scaled version of the control tibiae. Instead, the Longshanks tibia has become narrower in cross-section in relation to its increased length, leading to a decrease in overall bending strength when compared with control tibiae. These changes in bone shape and robusticity resemble the derived morphology of mammals adapted for running and jumping, with important implications for the material properties and strength of bone in these mammals.

Impacts of genetic correlation on the independent evolution of body mass and skeletal size in mammals.

BACKGROUND: Mammals show a predictable scaling relationship between limb bone size and body mass. This relationship has a genetic basis which likely evolved via natural selection, but it is unclear how much the genetic correlation between these traits in turn impacts their capacity to evolve independently. We selectively bred laboratory mice for increases in tibia length independent of body mass, to test the hypothesis that a genetic correlation with body mass constrains evolutionary change in tibia length. RESULTS: Over 14 generations, we produced mean tibia length increases of 9-13%, while mean body mass was unchanged, in selectively bred mice and random-bred controls. Using evolutionary scenarios with different selection and quantitative genetic parameters, we also found that this genetic correlation impedes the rate of evolutionary change in both traits, slowing increases in tibia length while preventing decreases in body mass, despite the latter's negative effect on fitness. CONCLUSIONS: Overall, results from this ongoing selection experiment suggest that parallel evolution of relatively longer hind limbs among rodents, for example in the context of strong competition for resources and niche partitioning in heterogeneous environments, may have occurred very rapidly on geological timescales, in spite of a moderately strong genetic correlation between tibia length and body mass.