PHOSPHO1 is a skeletal regulator of insulin resistance and obesity.

BACKGROUND: The classical functions of the skeleton encompass locomotion, protection and mineral homeostasis. However, cell-specific gene deletions in the mouse and human genetic studies have identified the skeleton as a key endocrine regulator of metabolism. The bone-specific phosphatase, Phosphatase, Orphan 1 (PHOSPHO1), which is indispensable for bone mineralisation, has been recently implicated in the regulation of energy metabolism in humans, but its role in systemic metabolism remains unclear. Here, we probe the mechanism underlying metabolic regulation by analysing Phospho1 mutant mice. RESULTS: Phospho1-/- mice exhibited improved basal glucose homeostasis and resisted high-fat-diet-induced weight gain and diabetes. The metabolic protection in Phospho1-/- mice was manifested in the absence of altered levels of osteocalcin. Osteoblasts isolated from Phospho1-/- mice were enriched for genes associated with energy metabolism and diabetes; Phospho1 both directly and indirectly interacted with genes associated with glucose transport and insulin receptor signalling. Canonical thermogenesis via brown adipose tissue did not underlie the metabolic protection observed in adult Phospho1-/- mice. However, the decreased serum choline levels in Phospho1-/- mice were normalised by feeding a 2% choline rich diet resulting in a normalisation in insulin sensitivity and fat mass. CONCLUSION: We show that mice lacking the bone mineralisation enzyme PHOSPHO1 exhibit improved basal glucose homeostasis and resist high-fat-diet-induced weight gain and diabetes. This study identifies PHOSPHO1 as a potential bone-derived therapeutic target for the treatment of obesity and diabetes.

Genome-wide association study of footrot in Texel sheep.

BACKGROUND: This is the first study based on a genome-wide association approach that investigates the links between ovine footrot scores and molecular polymorphisms in Texel sheep using the ovine 50 K SNP array (42 883 SNPs (single nucleotide polymorphisms) after quality control). Our aim was to identify molecular predictors of footrot resistance. METHODS: This study used data from animals selected from a footrot-phenotyped Texel sheep population of 2229 sheep with an average of 1.60 scoring records per animal. From these, a subset of 336 animals with extreme trait values for footrot was selected for genotyping based on their phenotypic records. De-regressed estimated breeding values (EBV) for footrot were used as pseudo-phenotypes in the genome-wide association analysis. RESULTS: Seven SNPs were significant on a chromosome-wise level but the association analysis did not reveal any genome-wise significant SNPs associated with footrot. Based on the current state of knowledge of the ovine genome, it is difficult to clearly link the function of the genes that contain these significant SNPs with a potential role in resistance/susceptibility to footrot. Linkage disequilibrium (LD) was analysed as one of the factors that influence the power of detecting QTL (quantitative trait loci). A mean LD of 0.20 (r(2) at a distance of 50 kb between two SNPs) in the population analysed was estimated. LD declined from 0.15 to 0.07 and to 0.04 at distances between two SNPs of 100, 1000 and 2000 kb, respectively. CONCLUSIONS: Based on a relatively small number of genotyped animals, this study is a first step to search for genomic regions that are involved in resistance to footrot using the ovine 50 K SNP array. Seven SNPs were found to be significant on a chromosome-wise level. No major genome-wise significant QTL were identified.

Limits to sustained energy intake. XXII. Reproductive performance of two selected mouse lines with different thermal conductance.

Maximal sustained energy intake (SusEI) appears limited, but the factors imposing the limit are disputed. We studied reproductive performance in two lines of mice selected for high and low food intake (MH and ML, respectively), and known to have large differences in thermal conductance (29% higher in the MH line at 21°C). When these mice raised their natural litters, their metabolisable energy intake significantly increased over the first 13 days of lactation and then reached a plateau. At peak lactation, MH mice assimilated on average 45.3% more energy than ML mice (222.9±7.1 and 153.4±12.5 kJ day(-1), N=49 and 24, respectively). Moreover, MH mice exported on average 62.3 kJ day(-1) more energy as milk than ML mice (118.9±5.3 and 56.6±5.4 kJ day(-1), N=subset of 32 and 21, respectively). The elevated milk production of MH mice enabled them to wean litters (65.2±2.1 g) that were on average 50.2% heavier than litters produced by ML mothers (43.4±3.0 g), and pups that were on average 27.2% heavier (9.9±0.2 and 7.8±0.2 g, respectively). Lactating mice in both lines had significantly longer and heavier guts compared with non-reproductive mice. However, inconsistent with the 'central limit hypothesis', the ML mice had significantly longer and heavier intestines than MH mice. An experiment where the mice raised litters of the opposing line demonstrated that lactation performance was not limited by the growth capacity of offspring. Our findings are consistent with the idea that the SusEI at peak lactation is constrained by the capacity of the mothers to dissipate body heat.

Genetic and genomic analyses of musculoskeletal differences between BEH and BEL strains.

Berlin high (BEH) and Berlin low (BEL) strains selected for divergent growth differ threefold in body weight. We aimed at examining muscle mass, which is a major contributor to body weight, by exploring morphological characteristics of the soleus muscle (fiber number and cross sectional area; CSA), by analyzing the transcriptome of the gastrocnemius and by initiating quantitative trait locus (QTL) mapping. BEH muscles were four to eight times larger than those of BEL. In substrain BEH+/+, mutant myostatin was replaced with a wild-type allele; however, BEH+/+muscles still were two to four times larger compared with BEL. BEH soleus muscle fibers were two times more numerous (P < 0.0001) and CSA was two times larger (P < 0.0001) compared with BEL. In addition, soleus femoral attachment anomaly (SFAA) was observed in all BEL mice. One significant (Chr 1) and four suggestive (Chr 3, 4, 6, and 9) muscle weight QTLs were mapped in a 21-day-old F2 intercross (n = 296) between BEH and BEL strains. The frequency of SFAA incidence in the F2 and in the backcross to BEL strain (BCL) suggested the presence of more than one causative gene. Two suggestive SFAA QTLs were mapped in BCL; however, their peak markers were not associated with the phenotype in F2. RNA-Seq analysis revealed 2,148 differentially expressed (P < 0.1) genes and 45,673 single nucleotide polymorphisms and >2,000 indels between BEH+/+ and BEL males. In conclusion, contrasting muscle traits and genomic and gene expression differences between BEH and BEL strains provide a promising model for the search for genes involved in muscle growth and musculoskeletal morphogenesis.

Analyses of muscle spindles in the soleus of six inbred mouse strains.

Adult muscle size and fibre-type composition are heritable traits that vary substantially between individuals. We used inbred mouse strains in which soleus muscle mass varied by an order of magnitude to explore whether properties of muscle spindles can also be influenced by genetic factors. Skip-serial cross-sections of soleus muscles dissected from 15 male mice of BEH, BEL, C57BL/6J, DUH, LG/J and SM/J strains were analysed for number of muscle spindles and characteristics of intrafusal and extrafusal fibres following ATPase staining. The BEL and DUH strains determined the range of: soleus mean size, a 10-fold difference from 2.1 to 22.3 mg, respectively; the mean number of extrafusal fibres, a 2.5-fold difference from 497 to 1249; and mean fibre-cross-sectional area, three-fold difference, e.g. for type 1 fibres, from 678 to 1948 µm². The range of mean proportion of type 1 fibres was determined by C57BL/6J (31%) and DUH (64%) strains. The mean number of spindles per muscle ranged between nine (LG/J) and 13 (BEL) (strain effect P < 0.02). Genetic correlations between spindle count and muscle weight or properties of extrafusal fibres were weak and not statistically significant. However, there was a strong correlation between the proportion of spindles with more than one bag2 fibre and the proportion of extrafusal fibres that were of type 1, and strain-dependent variation in the numbers of such spindles was statistically significant. The numbers of intrafusal fibres per spindle ranged from 2 to 8, with the most common complement of four found in 75.6% of spindles. There were no significant differences between the strains in the mean numbers of intrafusal fibres; however, the variance of the number was significantly less for the C57BL/6J strain than for any of the others. We conclude that abundance of muscle spindles and their intrafusal-fibre composition are substantially determined by genetic factors, which are different from those affecting muscle size and properties of the extrafusal fibres.

Stanniocalcin-2 inhibits skeletal muscle growth and is upregulated in functional overload-induced hypertrophy.

AIMS: Stanniocalcin-2 (STC2) has recently been implicated in human muscle mass variability by genetic analysis. Biochemically, STC2 inhibits the proteolytic activity of the metalloproteinase PAPP-A, which promotes muscle growth by upregulating the insulin-like growth factor (IGF) axis. The aim was to examine if STC2 affects skeletal muscle mass and to assess how the IGF axis mediates muscle hypertrophy induced by functional overload. METHODS: We compared muscle mass and muscle fiber morphology between Stc2-/- (n = 21) and wild-type (n = 15) mice. We then quantified IGF1, IGF2, IGF binding proteins -4 and -5 (IGFBP-4, IGFBP-5), PAPP-A and STC2 in plantaris muscles of wild-type mice subjected to 4-week unilateral overload (n = 14). RESULTS: Stc2-/- mice showed up to 10% larger muscle mass compared with wild-type mice. This increase was mediated by greater cross-sectional area of muscle fibers. Overload increased plantaris mass and components of the IGF axis, including quantities of IGF1 (by 2.41-fold, p = 0.0117), IGF2 (1.70-fold, p = 0.0461), IGFBP-4 (1.48-fold, p = 0.0268), PAPP-A (1.30-fold, p = 0.0154) and STC2 (1.28-fold, p = 0.019). CONCLUSION: Here we provide evidence that STC2 is an inhibitor of muscle growth upregulated, along with other components of the IGF axis, during overload-induced muscle hypertrophy.

The effect of gestational undernutrition on maternal weight change and fetal weight in lines of mice selected for different growth characteristics.

The present study investigated whether the genetic growth characteristics (fast or slow growing, lean or fat) of a mother influences her ability to partition nutrients to developing offspring. A total of sixty-one pregnant mice of three selected lines were used: fast-growing, relatively fat (FF, n 19); fast-growing, relatively lean (FL, n 23); and normal growth, relatively lean (NL, n 19). On day 1 of pregnancy, mice were given either ad libitum access to food (control (C): n 32) or pair-fed at 80 % of C intake (restricted (R): n 29). Feed intake and dam weight were measured daily. The weight of the mouse, organs, mammary tissue and the weight of fetuses and placentas were determined at day 18 of gestation. Overall, R dams gained less than half the weight of C dams during gestation. NL dams gained the most weight, and FF dams gained the least weight (P < 0·001). R dams in the fast-growing lines mobilised significantly more body fat during gestation than the NL line (P < 0·001) and had a greater reduction in mammary tissue growth. The relative weight of the litter increased in R dams of the FF line but was reduced in both the lean lines. Undernutrition reduced fetal and placental weight, and reduced placental efficiency in all the lines. The reduction was least in the FF line and greatest in the FL line. The data suggest that selection of animals for different growth characteristics alters their response to undernutrition during pregnancy, the relatively fat line was better able to buffer its offspring from the effects of undernutrition than the lean lines, regardless of their underlying rate of growth.

Interactive effects of protein nutrition, genetic growth potential and Heligmosomoides bakeri infection pressure on resilience and resistance in mice.

The ability of animals to cope with an increasing parasite load, in terms of resilience and resistance, may be affected by both nutrient supply and demand. Here, we hypothesized that host nutrition and growth potential interact and influence the ability of mice to cope with different parasite doses. Mice selected for high (ROH) or low (ROL) body weight were fed a low (40 g/kg; LP) or high (230 g/kg; HP) protein diet and infected with 0, 50, 100, 150, 200 or 250 L3 infective Heligmosomoides bakeri larvae. ROH-LP mice grew less at doses of 150 L3 and above, whilst growth of ROH-HP and of ROL mice was not affected by infection pressure. Total worm burdens reached a plateau at doses of 150L3, whilst ROH mice excreted fewer worm eggs than ROL mice. Serum antibodies increased with infection dose and ROH mice were found to have higher parasite-specific IgG1 titres than ROL mice. In contrast, ROL had higher total IgE titres than ROH mice, only on HP diets. The interaction between host nutrition and growth potential appears to differentially affect resilience and resistance in mice. However, the results support the view that parasitism penalises performance in animals selected for higher growth.

Genetic parameters associated with meat quality of Nellore cattle at different anatomical points of longissimus: Brazilian standards.

The present study estimated genetic parameters and evaluated the genetic and phenotypic correlations between meat quality characteristics of Nellore cattle evaluated at different anatomical points of the longissimus. Data from 1329 Nellore young bulls were used to evaluate, in the 5th and 12th ribs, marbling score (MAR), shear force (SF), cooking weight losses (CWL) and intramuscular fat (IMF). In addition, the subcutaneous fat thickness was measured at the 12th rib (SFT12) and between the last lumbar and the first sacral vertebrae (SFTLR), in the separation of loin and round. Results yielded moderate heritability coefficients for evaluated characteristics, except CWL. High genetic correlations (0.61) were found between measurements of SFT12 and SFTLR. MAR, IMF and SF were evaluated at the 5th and 12th rib. Meat quality and subcutaneous fat thickness measured at different anatomical points of the longissimus are genetically correlated and can be used in genetic selection programs to improve meat quality characteristics in Nellore cattle.

Predicting the shear value and intramuscular fat in meat from Nellore cattle using Vis-NIR spectroscopy.

Visible and near-infrared spectroscopy (Vis-NIRS) was tested for its effectiveness in predicting intramuscular fat (IMF) and WBSF in Nellore steers. Beef samples from longissimus thoracis, aged for either 2 or 7 days, had their spectra collected for wavelengths ranging from 400 to 1395 nm. Partial least squares regression models were developed for each trait. Determination coefficients of calibration models for WBSF ranged from 0.17 to 0.53. Considering WBSF in samples aged for 2 days, Vis-NIR correctly classified 100% of tough samples (>45 N), but wrongly classified all tender samples (≤45 N) as tough. Determination coefficients of calibration models for IMF ranged from 0.12 to 0.14. Vis-NIRS is a useful tool for identifying tough beef, but it is less effective in predicting tender samples and IMF. Additional studies are necessary to generate more robust models for the prediction of intramuscular fat in intact meat samples of Nellore cattle.

Effects of feed allowance and indispensable amino acid reduction on feed intake, growth performance and carcass characteristics of growing pigs.

The hypothesis that pigs placed on diets with reduced indispensable amino acid (AA) content attempts to offset the reduction in the nutrient density with increased feed intake was tested. In the experiment, feeds with a high or a low AA content were administrated to pigs fed ad-libitum or restrictively according to a 2 × 2 factorial design. Ninety-six barrows were housed in 8 pens (12 pigs/pen) equipped with automatic feeders. Within pen, and from 47 body weight (BW) onwards, 6 pigs were fed ad libitum. The others pigs were allowed to consume, as a maximum, the feed amounts indicated by the breeding company feeding plane to optimize the feed efficiency. In early (86-118 kg BW) and late (118-145 kg BW) finishing, the pigs of 4 pens received feeds with high indispensable AA contents (8.1 and 7.5 g lysine/kg in the two periods, respectively). The other pigs received feeds with reduced indispensable AA contents (lysine, methionine, threonine and tryptophan) by 9 and 18% in early and late finishing, respectively. Body lipid and protein (Pr) retentions were estimated from BW and back-fat depth measures recorded at the beginning and the end of each period. Nitrogen excretion was estimated as actual intake minus estimated N-retention (Pr/6.25). Pigs were slaughtered at 144 kg BW. Restricted feeding decreased feed intake (-7%), daily gain (-5%), carcass weight (-2.6%) and back-fat depth (-8.0%) but increased gain:feed ratio (+2%). The AA restriction increased feed intake (+5.9%), carcass weight (+4.9%) and intramuscular fat (+17.6%), and reduced carcass weight variation (-36%), with no effects on the feed efficiency and the estimated Pr (142 g/d). N excreted was reduced by feed (-9%) and dietary AA (-15%) restrictions. Irrespectively of the feeding level, the pigs responded to a reduction of the dietary essential AA content by increasing their feed intake.

Fine mapping of mouse QTLs for fatness using SNP data.

Quantitative trait loci (QTLs), as determined in crossbred studies, are a valuable resource to identify genes responsible for the corresponding phenotypic variances. Due to their broad chromosomal extension of some dozens of megabases, further steps are necessary to bring the number of candidate genes that underlie the detected effects to a reasonable order of magnitude. We use a set of 13,370 SNPs to identify informative haplotype blocks in 22 mouse QTLs for fatness. About half of the genes in a typical QTL overlap with haplotype blocks, which are different for the two base mouse lines, and which, thus, qualify for further analysis. For these genes we collect four more pieces of evidence for association with fat accumulation, namely (1) homology to genes identified in a Caenorhabditis elegans knock-out experiment as fat decreasing or fat increasing, (2) the overexpression of the genes in mouse fat, liver, muscle, or hypothalamus tissues, (3) the occurrence of a gene in several independently found QTLs, and (4) the information provided by gene ontology, to achieve a ranked list of 131 candidate genes. Ten genes fulfill three or four of the above sketched criteria and are discussed briefly, 121 further genes fulfilling two criteria are provided as on-line material. Viewing the genomic region of fatness-related QTLs under several different aspects is appropriate to assess the many thousands of genes that reside in such QTLs and to produce lists of more robust candidate genes.

A polygenic model of the metabolic syndrome with reduced circulating and intra-adipose glucocorticoid action.

Despite major advances in understanding monogenic causes of morbid obesity, the complex genetic and environmental etiology of idiopathic metabolic syndrome remains poorly understood. One hypothesis suggests that similarities between the metabolic disease of plasma glucocorticoid excess (Cushing's syndrome) and idiopathic metabolic syndrome results from increased glucocorticoid reamplification within adipose tissue by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD-1). Indeed, 11beta-HSD-1 is now a major therapeutic target. Because much supporting evidence for a role of adipose 11beta-HSD-1 comes from transgenic or obese rodents with single-gene mutations, we investigated whether the predicted traits of metabolic syndrome and glucocorticoid metabolism were coassociated in a unique polygenic model of obesity developed by long-term selection for divergent fat mass (Fat and Lean mice with 23 vs. 4% fat as body weight, respectively). Fat mice exhibited an insulin-resistant metabolic syndrome including fatty liver and hypertension. Unexpectedly, Fat mice had a marked intra-adipose (11beta-HSD-1) and plasma glucocorticoid deficiency but higher liver glucocorticoid action. Furthermore, metabolic disease was exacerbated only in Fat mice when challenged with exogenous glucocorticoids or a high-fat diet. Our data suggest that idiopathic metabolic syndrome might associate with such a novel pattern of glucocorticoid action and sensitivity in humans, with implications for tissue-specific therapeutic targeting of 11beta-HSD-1.

Baseline Muscle Mass Is a Poor Predictor of Functional Overload-Induced Gain in the Mouse Model.

Genetic background contributes substantially to individual variability in muscle mass. Muscle hypertrophy in response to resistance training can also vary extensively. However, it is less clear if muscle mass at baseline is predictive of the hypertrophic response. The aim of this study was to examine the effect of genetic background on variability in muscle mass at baseline and in the adaptive response of the mouse fast- and slow-twitch muscles to overload. Males of eight laboratory mouse strains: C57BL/6J (B6, n = 17), BALB/cByJ (n = 7), DBA/2J (D2, n = 12), B6.A-(rs3676616-D10Utsw1)/Kjn (B6.A, n = 9), C57BL/6J-Chr10A/J/NaJ (B6.A10, n = 8), BEH+/+ (n = 11), BEH (n = 12), and DUHi (n = 12), were studied. Compensatory growth of soleus and plantaris muscles was triggered by a 4-week overload induced by synergist unilateral ablation. Muscle weight in the control leg (baseline) varied from 5.2 ± 07 mg soleus and 11.4 ± 1.3 mg plantaris in D2 mice to 18.0 ± 1.7 mg soleus in DUHi and 43.7 ± 2.6 mg plantaris in BEH (p < 0.001 for both muscles). In addition, soleus in the B6.A10 strain was ~40% larger (p < 0.001) compared to the B6. Functional overload increased muscle weight, however, the extent of gain was strain-dependent for both soleus (p < 0.01) and plantaris (p < 0.02) even after accounting for the baseline differences. For the soleus muscle, the BEH strain emerged as the least responsive, with a 1.3-fold increase, compared to a 1.7-fold gain in the most responsive D2 strain, and there was no difference in the gain between the B6.A10 and B6 strains. The BEH strain appeared the least responsive in the gain of plantaris as well, 1.3-fold, compared to ~1.5-fold gain in the remaining strains. We conclude that variation in muscle mass at baseline is not a reliable predictor of that in the overload-induced gain. This suggests that a different set of genes influence variability in muscle mass acquired in the process of normal development, growth, and maintenance, and in the process of adaptive growth of the muscle challenged by overload.

Oxidative costs of reproduction in mouse strains selected for different levels of food intake and which differ in reproductive performance.

Oxidative damage caused by reactive oxygen species has been hypothesised to underpin the trade-off between reproduction and somatic maintenance, i.e., the life-history-oxidative stress theory. Previous tests of this hypothesis have proved equivocal, and it has been suggested that the variation in responses may be related to the tissues measured. Here, we measured oxidative damage (protein carbonyls, 8-OHdG) and antioxidant protection (enzymatic antioxidant activity and serum antioxidant capacity) in multiple tissues of reproductive (R) and non-reproductive (N) mice from two mouse strains selectively bred for high (H) or low (L) food intake, which differ in their reproductive performance, i.e., H mice have increased milk energy output (MEO) and wean larger pups. Levels of oxidative damage were unchanged (liver) or reduced (brain and serum) in R versus N mice, and no differences in multiple measures of oxidative protection were found between H and L mice in liver (except for Glutathione Peroxidase), brain or mammary glands. Also, there were no associations between an individual's energetic investment (e.g., MEO) and most of the oxidative stress measures detected in various tissues. These data are inconsistent with the oxidative stress theory, but were more supportive of, but not completely consistent, with the 'oxidative shielding' hypothesis.

Microarray gene expression analysis of the Fob3b obesity QTL identifies positional candidate gene Sqle and perturbed cholesterol and glycolysis pathways.

Obesity-related diseases are poised to become the primary cause of death in developed nations. While a number of monogenic causes of obesity have recently been identified, these are responsible for only a small proportion of human cases of obesity. Quantitative trait locus (QTL) studies using animal models have revealed hundreds of potential loci that affect obesity; however, few have been further analyzed beyond the original QTL scan. We previously mapped four QTL in an F(2) between divergently selected Fat (F) and Lean (L) lines. A QTL of large effect on chromosome 15 (Fob3) was subsequently mapped to a higher resolution into two smaller-effect QTL (Fob3a and Fob3b) using crosses between the F-line and a congenic line containing L-line alleles at the Fob3 QTL region. Here we report the gene expression characterization of Fob3b. Microarray expression analysis using the NIA-NIH 15K cDNA array set containing 14,938 mouse ESTs was employed to identify candidate genes and pathways that are differentially expressed between the F-line and a congenic line containing only the Fob3b QTL (Fob3b-line). Our study suggests squalene epoxidase (Sqle), a cholesterol biosynthesis enzyme, as a strong positional candidate gene for Fob3b. Several other cholesterol biosynthesis pathway genes unlinked to Fob3b were found to be differentially expressed, suggesting that a perturbation of this pathway could be in part responsible for the phenotypic difference between the F-line and Fob3b-line mice.

Long-term divergent selection on fatness in mice indicates a regulation system independent of leptin production and reception.

Divergent selection in mice on fatness over 60 generations produced a fat (F) and a lean (L) line, having about 22% and 4% body fat, respectively. To elucidate the importance of the leptin regulatory feedback loop in the genetic changes produced by this selection, Lep(ob) and Lepr(db) mutations causing leptin production and leptin receptor deficiency, respectively, were introgressed individually into both lines by repeated backcrossing. The fat amount increased significantly in homozygotes for Lep(ob) or Lepr(db) in both lines, for example, in F and L males from 8.5 to 18.8 and 17.2 g (P<0.001) and from 1.25 to 18.0 and 12.7 g (P<0.001), respectively. Line differences were, however, mostly maintained after introgression. Concentrations of circulating leptin were relatively independent of the original lines but heavily dependent on the introgressed genotype. Introgression of leptin production and receptor deficiencies had separate effects from long-term selection, indicating that the genes responsible for the line divergence must act independently of the leptin regulatory system. Energy budget analysis indicated that the major line differences were in the level of energy expended on physical activity, and these differences were preserved following introgression, suggesting that multiple pathways regulate fatness, which may be independently responsive to intervention.

Myostatin dysfunction impairs force generation in extensor digitorum longus muscle and increases exercise-induced protein efflux from extensor digitorum longus and soleus muscles.

Myostatin dysfunction promotes muscle hypertrophy, which can complicate assessment of muscle properties. We examined force generating capacity and creatine kinase (CK) efflux from skeletal muscles of young mice before they reach adult body and muscle size. Isolated soleus (SOL) and extensor digitorum longus (EDL) muscles of Berlin high (BEH) mice with dysfunctional myostatin, i.e., homozygous for inactivating myostatin mutation, and with a wild-type myostatin (BEH+/+) were studied. The muscles of BEH mice showed faster (P < 0.01) twitch and tetanus contraction times compared with BEH+/+ mice, but only EDL displayed lower (P < 0.05) specific force. SOL and EDL of age-matched but not younger BEH mice showed greater exercise-induced CK efflux compared with BEH+/+ mice. In summary, myostatin dysfunction leads to impairment in muscle force generating capacity in EDL and increases susceptibility of SOL and EDL to protein loss after exercise.

Factors affecting dystocia and offspring vigour in different sheep genotypes.

Birth difficulty and poor lamb vigour are significant causes of perinatal lamb mortality. In this study we investigated whether sheep breeds differing in appearance, muscularity and selection history also had differences in dystocia and lamb vigour, and considered some of the factors that may contribute to the variation in these traits. Data were collected at birth from a total of 3252 lambs of two terminal sire breeds selected for lean growth (Suffolk [S], n=500 and Texel [T], n=1207), from a Hill breed (Scottish Blackface [B], n=610), which has been mainly selected for hardiness, and a crossbred (Mule×T [M], n=935) representing a maternal line. For each lamb the degree of assistance at delivery, lamb presentation, amount of assistance to achieve successful sucking, sex, litter size and birth weight were recorded. T lambs required the most, and B and M lambs the least assistance at birth, S lambs were intermediate (% lambs assisted: T=55.7, S=30.7, B=22.7, M=24.9, P<0.001). T and S lambs were equally likely to be malpresented at birth (29% of births) and more likely to be malpresented than B or M lambs (20%; P<0.001). In T and S breeds lambs requiring veterinary assistance at delivery were mainly heavy and singleton lambs, whereas in B and M breeds these were exclusively low birth weight lambs in multiple litters. Although heavier lambs needed greater birth assistance, T lambs were lighter than S and M lambs, but heavier than B lambs (birth weight (kg): S=4.66, M=4.56, T=4.32, B=3.67, P<0.001). S lambs were more likely to require assistance with sucking than other breeds, and T lambs also required more assistance than B or M lambs (% lambs assisted to suck: S=56.0, T=31.6, M=19.8, B=18.4, P<0.001). Heavier lambs were more likely to suck unaided than lighter lambs (P<0.001). The data suggest that the two terminal sire breeds, selected narrowly for greater productivity (muscle growth and conformation), are more likely to experience birth difficulty and poorer lamb vigour than the breed selected for hardiness, or the cross breed. Whether these effects arise as a consequence of genetic selection (e.g. for specific lamb conformation), or as a result of management practices to achieve selection goals (e.g. increased intervention at lambing) is unknown. Specific actions to improve birth difficulty and lamb vigour, such as including these traits in the selection index, would be beneficial in improving the welfare of ewes and lambs of the terminal sire breeds.

Parallel selection mapping using artificially selected mice reveals body weight control loci.

Understanding how polygenic traits evolve under selection is an unsolved problem, because challenges exist for identifying genes underlying a complex trait and understanding how multilocus selection operates in the genome. Here we study polygenic response to selection using artificial selection experiments. Inbred strains from seven independent long-term selection experiments for extreme mouse body weight ("high" lines weigh 42-77 g versus 16-40 g in "control" lines) were genotyped at 527,572 SNPs to identify loci controlling body weight. We identified 67 parallel selected regions (PSRs) where high lines share variants rarely found among the controls. By comparing allele frequencies in one selection experiment against its unselected control, we found classical selective sweeps centered on the PSRs. We present evidence supporting two G protein-coupled receptors GPR133 and Prlhr as positional candidates controlling body weight. Artificial selection may mimic natural selection in the wild: compared to control loci, we detected reduced heterozygosity in PSRs in unusually large wild mice on islands. Many PSRs overlap loci associated with human height variation, possibly through evolutionary conserved functional pathways. Our data suggest that parallel selection on complex traits may evoke parallel responses at many genes involved in diverse but relevant pathways.