Evolution of hindlimb bone dimensions and muscle masses in house mice selectively bred for high voluntary wheel‐running behavior

We have used selective breeding with house mice to study coadaptation of morphology and physiology with the evolution of high daily levels of voluntary exercise. Here, we compared hindlimb bones and muscle masses from the 11th generation of four replicate High Runner (HR) lines of house mice bred for high levels of voluntary wheel running with four non‐selected control (C) lines. Mass, length, diameter, and depth of the femur, tibia‐fibula, and metatarsal bones, as well as masses of gastrocnemius and quadriceps muscles, were compared by analysis of covariance with body mass or body length as the covariate. Mice from HR lines had relatively wider distal femora and deeper proximal tibiae, suggesting larger knee surface areas, and larger femoral heads. Sex differences in bone dimensions were also evident, with males having thicker and shorter hindlimb bones when compared with females. Several interactions between sex, linetype, and/or body mass were observed, and analyses split by sex revealed several cases of sex‐specific responses to selection. A subset of the HR mice in two of the four HR lines expressed the mini‐muscle phenotype, characterized mainly by an ～50% reduction in hindlimb muscle mass, caused by a Mendelian recessive mutation, and known to have been under positive selection in the HR lines. Mini‐muscle individuals had elongated distal elements, lighter and thinner hindlimb bones, altered 3rd trochanter muscle insertion positions, and thicker tibia‐fibula distal widths. Finally, several differences in levels of directional or fluctuating asymmetry in bone dimensions were observed between HR and C, mini‐ and normal‐muscled mice, and the sexes. This study demonstrates that skeletal dimensions and muscle masses can evolve rapidly in response to directional selection on locomotor behavior.     

突变体——侏儒小鼠(dw~t)与津白3(TA3)不同日龄骨骼、体重及体长的比较

侏儒小鼠与津白3体重的比值为1:2.5。它们的骨骼、休重、休长及尾长的测量均具非常显著或显著意义。经均衡性检验,不同日龄、不同性别间TA3/dw~t小鼠的不同骨骼、体重、体长及尾长的比值变化是均匀一致的。证明侏儒小鼠骨骼、体重、体长及尾长均系均匀缩小,是身体各部分匀称的侏儒小鼠。     

Functional assessment of the muscle-bone unit in the lower leg

Based on the mechanostat theory and the muscle-bone hypothesis, a methodological assessment of the musculoskeletal status in health and disease should relate maximum muscle force in relation to bone mass and geometry. While useful (i.e. three-dimensional) measures of tibial bone parameters can be obtained by peripheral quantitative computed tomography (pQCT), intrinsic plantarflexor muscle force cannot be directly measured under in vivo condition in humans. Instead, tissue size, torque and ground reaction force have been used as proxy markers of intrinsic muscle force. However, most of these proxy markers are not or insufficiently representative of maximum force. Based on our recent research, we describe a novel approach for the assessment of the lower leg muscle-bone unit in health and disease. It incorporates multiple one-legged hopping (m1LH) to assess maximum voluntary ground reaction force acting on the forefoot (F(m1LH)) and bone mineral content at the 14%-site of tibia length (vBMC(14%)) as assessed by pQCT. Using the quantitative relationship between these two variables in conjunction with F(m1LH) per body weight, we present a two-step quantitative diagnostic algorithm to discriminate between primary and secondary bone disorders in children and adults.     

A mammary adenocarcinoma murine model suitable for the study of cancer immunoediting

Background

Cancer immunoediting is a dynamic process composed of three phases: elimination (EL), equilibrium (EQ) and escape (ES) that encompasses the potential host-protective and tumor-sculpting functions of the immune system throughout tumor development. Animal models are useful tools for studying diseases such as cancer. The present study was designed to characterize the interaction between mammary adenocarcinoma M-406 and CBi, CBi⁻ and CBi/L inbred mice lines.

Results

The mammary adenocarcinoma M-406 developed spontaneously in a CBi mouse. CBi/L and CBi⁻ mice were artificially selected for body conformation from CBi. When CBi mice are s.c. challenged with M-406, tumor growths exponentially in 100% of animals, while in CBi⁻ the tumor growths briefly and then begins a rejection process in 100% of the animals. In CBi/L the growth of the tumor shows the three phases: 51.6% in ES, 18.5% in EQ and 29.8% in EL.

Conclusions

The results obtained support the conclusion that the system M-406 plus the inbred mouse lines CBi, CBi⁻ and CBi/L, is a good murine model to study the process of tumor immunoediting.     

Disproportional skeletal growth and markedly decreased bone mineral content in growth hormone receptor -/- mice

Growth hormone (GH) is important for skeletal growth as well as for a normal bone metabolism in adults. The skeletal growth and adult bone metabolism was studied in mice with an inactivated growth hormone receptor (GHR) gene. The lengths of femur, tibia, and crown-rump were, as expected, decreased in GHR-/- mice. Unexpectedly, GHR-/- mice displayed disproportional skeletal growth reflected by decreased femur/crown-rump and femur/tibia ratios. GHR-/- mice demonstrated decreased width of the growth plates in the long bones and disturbed ossification of the proximal tibial epiphysis. Furthermore, the area bone mineral density (BMD) as well as the bone mineral content (BMC)/body weight were markedly decreased in GHR-/- mice. The decrease in BMC in GHR-/- mice was not due to decreased trabecular volumetric BMD but to a decreased cross-sectional cortical bone area In conclusion, GHR-/- mice demonstrate disproportional skeletal growth and markedly decreased bone mineral content.     

Influence of early qualitative feed restriction and environmental temperature on long bone development of broiler chickens

This investigation was carried out to study the influence of early qualitative feed restriction and environmental rearing temperature on long bone development in broiler. Energy and protein restriction reduced femur width and humerus weight, but did not affect tibia parameters. Broilers kept at cold environmental temperature showed reduced femur, tibia and humerus length and tibia weight, but the calculated density was not affected by rearing temperature. These findings suggest that qualitative feed restriction and environmental temperature influenced the normal long bone growth; however, bone weight/bone length index (calculated density) was not affected by rearing temperature.     

Analysis of the independent power of age-related,anthropometric and mechanical factors as determinants of the structure of radius and tibia in normal adults. A pQCT study

To compare the independent influence of mechanical and non-mechanical factors on bone features, multiple regression analyses were performed between pQCT indicators of radius and tibia bone mass, mineralization, design and strength as determined variables, and age or time since menopause (TMP), body mass, bone length and regional muscles’ areas as selected determinant factors, in Caucasian, physically active, untrained healthy men and pre- and post-menopausal women. In men and pre-menopausal women, the strongest influences were exerted by muscle area on radial features and by both muscle area and bone length on the tibia. Only for women, was body mass a significant factor for tibia traits. In men and pre-menopausal women, mass/design/strength indicators depended more strongly on the selected determinants than the cortical vBMD did (p<0.01-0.001 vs n.s.), regardless of age. However, TMP was an additional factor for both bones (p<0.01-0.001). The selected mechanical factors (muscle size, bone lengths) were more relevant than age/TMP or body weight to the development of allometrically-related bone properties (mass/design/strength), yet not to bone tissue “quality” (cortical vBMD), suggesting a determinant, rather than determined role for cortical stiffness. While the mechanical impacts of muscles and bone levers on bone structure were comparable in men and pre-menopausal women, TMP exerted a stronger impact than allometric or mechanical factors on bone properties, including cortical vBMD.     

Divergent selection on 63-day body weight in the rabbit: response on growth,carcass and muscle traits

The effects of selection for growth rate on weights and qualitative carcass and muscle traits were assessed by comparing two lines selected for live body weight at 63 days of age and a cryopreserved control population raised contemporaneously with generation 5 selected rabbits. The animals were divergently selected for five generations for either a high (H line) or a low (L line) body weight, based on their BLUP breeding value. Heritability (h²) was 0.22 for 63-d body weight (N = 4754). Growth performance and quantitative carcass traits in the C group were intermediate between the H and L lines (N = 390). Perirenal fat proportion (h² = 0.64) and dressing out percentage (h² = 0.55) ranked in the order L < H = C (from high to low). The weight and cross-sectional area of the Semitendinosus muscle, and the mean diameter of the constitutive myofibres were reduced in the L line only (N = 140). In the Longissimus muscle (N = 180), the ultimate pH (h² = 0.16) and the maximum shear force reached in the Warner-Braztler test (h² = 0.57) were slightly modified by selection.     

Fine mapping dissects pleiotropic growth quantitative trait locus into linked loci

A recurring issue in studies of quantitative trait loci (QTLs) is whether QTLs that appear to have pleiotropic effects are indeed caused by pleiotropy at single loci or by linked QTLs. Previous work identified a QTL that affected tail length in mice and the lengths of various bones, including the humerus, ulna, femur, tibia, and mandible. The effect of this QTL on tail length has since been found to be due to multiple linked QTLs and so its apparently pleiotropic effects may have been due to linked QTLs with distinct effects. In the present study we examined a line of mice segregating only for a 0.94-Mb chromosomal region known to contain a subset of the QTLs influencing tail length. We measured a number of skeletal dimensions, including the lengths of the skull, mandible, humerus, ulna, femur, tibia, calcaneus, metatarsus, and a tail bone. The QTL region was found to have effects on the size of the mandible and length of the tail bone, with little or no effect on the other traits. Using a randomization approach, we rejected the null hypothesis that the QTL affected all traits equally, thereby demonstrating that the pleiotropic effects reported earlier were due to linked loci with distinct effects. This result underlines the possibility that seemingly pleiotropic effects of QTLs may frequently be due to linked loci and that high-resolution mapping will often be required to distinguish between pleiotropy and linkage.     

Effect of initial size on subsequent growth and carcass characteristics of divergently selected channel catfish

Summary Two experiments were conducted simultaneously to determine (1) if fast-growing fingerlings of channel catfish, Ictalurus punctatus, could be identified by simple visual selection of body size and (2) if initial size advantages influenced subsequent growth and carcass traits of divergently selected channel catfish. Exp. 1 included large (L), medium (M), and small (S) fingerling sizes from each of the control (C), selected upward (+) and selected downward (–) lines for body weight. Exp. 2 included all fmgerlings of the same size (25±5 g) from the 3 lines. Catfish from the L size-class, within each full-sib family in each line, were consistently heavier and longer than M and S size-classes throughout the 53-week experimental period. Fingerlings from the M size-class were also superior in growth to those from the S size-class. Catfish from the + line exceeded those from the C and –lines in body weight and total length at the conclusion of Exp. 1 but not in Exp. 2. This was attributed to the selection of equal size fmgerlings in Exp. 2 which may have excluded fingerlings with the best growth potential from the + body weight line. Results of the two experiments combined indicated that one generation of divergent selection has created genetic differences among lines of channel catfish.Supported by State and Hatch funds allocated to the Georgia Agricultural Experiment Station     

Type I diabetic bone phenotype is location but not gender dependent

Bone is highly dynamic and responsive. Bone location, bone type and gender can influence bone responses (positive, negative or none) and magnitude. Type I diabetes induces bone loss and increased marrow adiposity in the tibia. We tested if this response exhibits gender and location dependency by examining femur, vertebrae and calvaria of male and female, control and diabetic BALB/c mice. Non-diabetic male mice exhibited larger body, muscle, and fat mass, and increased femur BMD compared to female mice, while vertebrae and calvarial bone parameters did not exhibit gender differences. Streptozotocin-induced diabetes caused a reduction in BMD at all sites examined irrespective of gender. Increased marrow adiposity was evident in diabetic femurs and calvaria (endochondrial and intramembranous formed bones, respectively), but not in vertebrae. Leptin-deficient mice also exhibit location dependent bone responses and we found that serum leptin levels were significantly lower in diabetic compared to control mice. However, in contrast to leptin-deficient mice, the vertebrae of T1-diabetic mice exhibit bone loss, not gain. Taken together, our findings indicate that TI-diabetic bone loss in mice is not gender, bone location or bone type dependent, while increased marrow adiposity is location dependent.     

Body weight and tail length divergence in mice selected for rate of development

A series of mouse lines has been produced by 19 generations of restricted index selection for rate of development during early and late ontogeny. The selection program was based on an index with the following four replicated selection treatments: E(+) and E(-) were selected to alter birth to 10-day body weight gain while holding late gain for both selection lines constant; correspondingly, L(+) and L(-) were selected to alter 28- to 56-day body weight gain holding early gain for both lines constant. Herein, we characterize response to selection for growth rate by analyzing age-specific mouse body weight and tail lengths and for growth curves using a logistics model. Selection on developmental rate has resulted in divergence in both age-specific and growth curve traits. E(+) and L(+) lines reached identical weights during the late selection interval, then diverged to unique mature weights. E(-) and L(-) lines similarly achieved identical weights during late selection and diverged to unique mature weights. However, the shapes of early and late growth curves were significantly divergent, and at least two distinct growth patterns are shown to result from selection. Response in body weight gain was accompanied by similar, though less pronounced, change in tail length traits. Significant response during intervals of restricted growth was also found, especially in lines selected for late gain. The evolution of the growth trajectory under restricted index selection is discussed in terms of drift and available additive genetic variation and covariation.     

Sexually dimorphic male horns and their use in agonistic behaviors in the horn-headed cricket <Emphasis Type="Italic">Loxoblemmus doenitzi</Emphasis> (Orthoptera: Gryllidae)

Sexual dimorphism, the difference between the sexes in secondary sexual characters, is in general driven by processes of sexual selection. The horn-headed cricket, Loxoblemmus doenitzi, exhibits sexual dimorphism in head shape. Males have flat heads and triangular horns on both sides of their heads, whereas females have rounded heads and no horns. We hypothesized that male horns have evolved due to intra-sexual selection, in which males use these horns as weapons in aggressive interactions. We tested two predictions of this hypothesis by conducting agonistic trials with field-caught males of L. doenitzi: (1) the horns should be used in agonistic interactions between males, and (2) the asymmetry in horn size or horn use may determine contest outcome. Horn length was significantly correlated with thorax length and hind femur length. During agonistic interactions, males aggressively used their horns by beating the opponent’s horns with their own or by poking the opponent’s body. However, logistic regression analysis revealed that neither horn length nor horn use were significant factors for contest outcome. Instead, body size was significant for determining contest outcome. We discuss possible scenarios for evolution of male horns in L. doenitzi.     

Temperature preference differences between strains of Mus musculus,associated variables,and ecological implications

Summary Temperature preference differences were found between three inbred strains of mice, Balb/c having a lower preference than C3H and C57Bl. Hybrids with Balb/c had preferences between the inbred strains, while the C3HxC57Bl hybrid was close to the component inbred strains. Mice choosing low temperatures were heavier, had higher body temperatures and lower hair densities than those choosing high temperatures. Arguments are presented for regarding these associations as adaptive, and being morphological and physiological correlates of the behavioural measure temperature preference, a trait itself of ecological significance in habitat selection. It is considered that these traits are subjected to stabilizing selection for the optimum for a given population, to provide the most efficient habitat selection for that population.Tail length is not so intimately associated with temperature preference and shows overdominance in the direction of long tails unlike the other traits measured. In any case, published data show that while there is a general tendency for shorter tails to occur in environments with low temperatures and vice versa, this is not always true, so that tail length is not necessarily a good guide to the habitat selected.Mice were measured at two ages (15 to 18 days, and 55 to 58 days) for temperature preference, body weight and tail length. Developmental changes were found in that the preferred temperature was lower in the older mice. Furthermore, relative changes between age groups for all traits differed between some genotypes, indicating varying developmental changes according to genotype.     

Characterization of a QTL affecting skeletal size in mice

Abstract Previous work identified a tail length QTL on Chromosome (Chr) 1 in an F₂ population of C57BL/6J × DBA/2J mice. The goals of the present study were to (1) refine the position of this QTL by additional genotyping of samples from the original study; (2) confirm the effect of this QTL by producing a partially congenic strain carrying the C57BL/6J allele against the DBA/2J background; and (3) examine the effect of the QTL on skeletal dimensions. The presence of the QTL was confirmed in a new F₂ population (N = 431) derived from the partially congenic strain, and estimates of its additive effects were similar to those from the original F₂ population (N = 901) in both sexes, i.e., the C57BL/6J chromosomal segment increased tail length, the additive effect (half the difference between homozygotes) being 0.5–0.8 standard deviations. The QTL region was more than halved, relative to that in the previous study, to an 8-cM region between D1Mit30 and D1Mit57. Among a subsample of individuals (N = 30) from the new F₂ population that were not recombinant within the QTL region, there was a significant additive effect of the QTL on the length of the humerus, femur, tibia, mandible, scapula, pelvic girdle, and a tail bone; the direction of the effect was the same as for tail length. No significant effect was found on the number of bones in the tail or on the dimensions of the ulna, skull, or first vertebra.     

Brief communication: Prenatal and early postnatal stress exposure influences long bone length in adult rat offspring

Stress during the prenatal and early postnatal periods (perinatal stress, PS) is known to impact offspring cognitive, behavioral, and physical development, but effects on skeletal growth are not clear. Our objective was to analyze effects of variable, mild, daily PS exposure on adult offspring long bone length. Twelve pregnant rat dams were randomly assigned to receive variable stress from gestational days 14–21 (Prenatal group), postpartum days 2–9 (Postnatal), both periods (Pre–Post), or no stress (Control). Differences in adult offspring tibia and femur length were analyzed among treatment groups. Mean tibia length differed among groups for males (P = 0.016) and females (P = 0.009), and differences for femur length approached significance for males (P = 0.051). Long bone length was shorter among PS‐exposed offspring, especially those exposed to postnatal stress (Postnatal and Pre–Post groups). Results persisted when controlling for nose–tail length. These differences might reflect early stunting that is maintained in adulthood, or delayed growth among PS‐exposed offspring. This study suggests that PS results in shorter long bones in adulthood, independently of effects on overall body size. Stunting and growth retardation are major global health burdens. Our study adds to a growing body of evidence suggesting that PS is a risk factor for poor linear growth. Am J Phys Anthropol 149:307–311, 2012. © 2012 Wiley Periodicals, Inc.     

Bone length and muscle weight in mice subjected to genetic selection for the relative length of the tibia and radius

Mice which had undergone 5 generations of selection for high and low values of the tibia length/radius length ratio were compared with unselected controls at 10 weeks of age. The ratio responded to selection in both directions. The length of the radius was increased in the low line while the response in the high line was due to an increase in the length of the tibia and a small but statistically insignificant decrease in radius length. High and low line mice were heavier than the controls. The responses of the tibia and femur and of the radius and humerus were generally similar, suggesting the existence of an association between the genetic control of the lengths of the bones of a limb. There were also correlated responses in the weight of the tibialis anterior, biceps brachii and sternomastoid, and there is some evidence to suggest that the weight of a muscle may be influenced by the length of a bone to which it is attached.     

Effects of spirulina, a blue-green alga, on bone metabolism in ovariectomized rats and hindlimb-unloaded mice

The safety and effectiveness were examined of the spirulina alga on bone metabolism in ovariectomized estrogen-deficient rats and hindlimb-unloaded mice. The dosage range was from an amount equal to that recommended in so-called health foods for humans (0.08 g/kg BW/day) to a 100-fold higher dose. The bone mineral density (BMD) of the whole femur and tibia of ovariectomized rats in the any spirulina-treated groups was not significantly different from that of the ovariectomized group, although BMD of the distal femur and proximal tibia was significantly lower in the spirulina-treated groups than in the ovariectomized group after a 6 week-experimental period. BMD of the femur and tibia was not affected by treatment with any dose of spirulina in hindlimb-unloaded mice. These results suggest that the intake of spirulina decreased BMD in the trabecular bone of rodents under estrogen-deficient conditions.     

Differential effects of jump versus running exercise on trabecular architecture during remobilization after suspension-induced osteopenia in growing rats

High-impact exercise is considered to be very beneficial for bones. We investigated the ability of jump exercise to restore bone mass and structure after the deterioration induced by tail suspension in growing rats and made comparisons with treadmill running exercise. Five-week-old male Wistar rats (n = 28) were randomly assigned to four body weight-matched groups: a spontaneous recovery group after tail suspension (n = 7), a jump exercise group after tail suspension (n = 7), a treadmill running group after tail suspension (n = 7), and age-matched controls without tail suspension or exercise (n = 7). Treadmill running was performed at 25 m/min, 1 h/day, 5 days/wk. The jump exercise protocol consisted of 10 jumps/day, 5 days/wk, with a jump height of 40 cm. Bone mineral density (BMD) of the total right femur was measured by dual-energy X-ray absorptiometry. Three-dimensional trabecular bone architecture at the distal femoral metaphysis was evaluated using microcomputed tomography. After 5 wk of free remobilization, right femoral BMD, right hindlimb muscle weight, and body weight returned to age-matched control levels, but trabeculae remained thinner and less connected. Although both jump and running exercises during the remobilization period increased trabecular bone mass, jump exercise increased trabecular thickness, whereas running exercise increased trabecular number. These results indicate that restoration of trabecular bone architecture induced by jump exercise during remobilization is predominantly attributable to increased trabecular thickness, whereas running adds trabecular bone mass through increasing trabecular number, and suggest that jumping and running exercises have different mechanisms of action on structural characteristics of trabecular bone.     

Shape and size of the body vs. musculoskeletal stress markers

The objective of this paper is to assess the relationship between the degree of development of muscle attachment sites (musculoskeletal stress markers - MSM1) and the length and circumference measurements of long bones and the body build expressed with the reconstructed values of body height (BH) and body mass (BM). The bone material (102 male and 99 female skeletons) used in the study was collected in the medieval burial ground in Cedynia, Poland. The authors analyzed 10 musculoskeletal stress markers located on the scapula (2), humerus (2), radius (2), femur (2) and tibia (2). The frequency and the degree of expression of muscle attachment size was carried out using the scale prepared by Myszka (2007). The scale encompassed three degrees of expression of muscle attachment size. Only changes of robusticity type (nonpathological changes) were taken into account. The assessment of body build of individuals was carried out according to the method proposed by Vancata & Charvátová (2001). Body height was reconstructed from the length of the humerus and femur using eight equations. Body mass was reconstructed from the measurements of the breadth of the proximal and distal sections of the femur and tibia (mechanical method) using twenty one equations. The equations were developed for different reference populations. The same equations were used for men and women. The correlation between the MSM and the length and circumference measurements of the bones was analyzed using the principal components analysis and the Gamma correlation coefficient. The strength of the correlation between the reconstructed body build traits (BH, BM) and the moderate degree of musculoskeletal stress markers expression was studied based on the principal components method and the Pearson correlation coefficient. A linear correlation was found between musculoskeletal stress markers and the circumference measurements and the reconstructed body mass, but no relationship with body height and the length measurements of long bones was revealed. From previous research it is evident that the relationship between the MSM and metric skeletal traits does not occur in every population. Divergent findings necessitate further corroboration of results on diverse skeletal material.