J. Timothy Lightfoot

Quantitative trait loci for physical activity traits in mice

The genomic locations and identities of the genes that regulate voluntary physical activity are presently unknown. The purpose of this study was to search for quantitative trait loci (QTL) that are linked with daily mouse running wheel distance, duration, and speed of exercise. F(2) animals (n = 310) derived from high active C57L/J and low active C3H/HeJ inbred strains were phenotyped for 21 days. After phenotyping, genotyping with a fully informative single-nucleotide polymorphism panel with an average intermarker interval of 13.7 cM was used. On all three activity indexes, sex and strain were significant factors, with the F(2) animals similar to the high active C57L/J mice in both daily exercise distance and duration of exercise. In the F(2) cohort, female mice ran significantly farther, longer, and faster than male mice. QTL analysis revealed no sex-specific QTL but at the 5% experimentwise significance level did identify one QTL for duration, one QTL for distance, and two QTL for speed. The QTL for duration (DUR13.1) and distance (DIST13.1) colocalized with the QTL for speed (SPD13.1). Each of these QTL accounted for approximately 6% of the phenotypic variance, whereas SPD9.1 (chromosome 9, 7 cM) accounted for 11.3% of the phenotypic variation. DUR13.1, DIST13.1, SPD13.1, and SPD9.1 were subsequently replicated by haplotype association mapping. The results of this study suggest a genetic basis of voluntary activity in mice and provide a foundation for future candidate gene studies.

Interstrain variation in murine aerobic capacity.

PURPOSE The contribution of genetic factors to aerobic capacity is unknown. The purpose of this study was to measure maximal aerobic performance among inbred strains of mice to provide basic heritability estimates. METHODS Eight female mice, 8 to 10 wk old, in 10 inbred strains (A/J, AKR/J, Balb/cJ, C(3)H/HeJ, C57Bl/6J, C57L/J, C(3)Heb/FeJ, CBA/J, DBA/2J, and SWR/J) were run on a treadmill until exhaustion. The protocol started at 22 m.min(-1) and increased in speed approximately 6 m.min(-1) every 4 min. After 4 min at 42.4 m.min(-1), the grade was increased 2% every 4 min thereafter until the mouse could not run off of the shock grid (150 V; 1.5 mA). RESULTS There were significant differences between inbred strains in maximal duration of exercise accomplished (P < 0.0001). The order of strain-specific exercise duration was Balb/cJ > SWR/J > CBA/J > C57L/J > C3H/HeJ > C3Heb/FeJ > C57Bl/6J > AKR/J > DBA/2J > A/J. Two measures of heritability in the broad sense, intraclass correlation (0.73), and the coefficient of genetic determination (0.58) were both significant. CONCLUSION These data indicate that there is a strong genetic contribution to aerobic capacity in mice.

Altered dopaminergic profiles: implications for the regulation of voluntary physical activity

The biological regulating factors of physical activity in animals are not well understood. This study investigated differences in the central mRNA expression of seven dopamine genes (Drd1, Drd2, Drd3, Drd4, Drd5, TH, and DAT) between high active C57/LJ (n=17) male mice and low active C3H/HeJ (n=20) male mice, and between mice with access to a running wheel and without running wheel access within strain. Mice were housed with running wheels interfaced with a computer for 21 days with distance and duration recorded every 24 h. On day 21, the striatum and nucleus accumbens were removed during the active period (approximately 9 pm) for dopaminergic analysis. On average, the C57L/J mice with wheels ran significantly farther (10.25+/-1.37 km/day vs. 0.01+/-0.09 km/day, p<0.001), longer (329.73+/-30.52 min/day vs. 7.81+/-6.32 min/day, p<0.001), and faster (31.27+/-3.13 m/min vs. 11.81+/-1.08 m/min, p<0.001) than the C3H/HeJ mice with wheels over the 21 day period. No differences in gene expression were found between mice in either strain with wheels and those without wheels suggesting that access to running wheels did not alter dopaminergic expression. In contrast, relative expression for two dopamine genes was significantly lower in the C57L/J mice compared to the C3H/HeJ mice. These results indicate that decreased dopaminergic functioning is correlated with increased activity levels in C57L/J mice and suggests that D1-like receptors as well as tyrosine hydroxylase (an indicator of dopamine production), but not D2-like receptors may be associated with the regulation of physical activity in inbred mice.

Strain screen and haplotype association mapping of wheel running in inbred mouse strains.

Previous genetic association studies of physical activity, in both animal and human models, have been limited in number of subjects and genetically homozygous strains used as well as number of genomic markers available for analysis. Expansion of the available mouse physical activity strain screens and the recently published dense single-nucleotide polymorphism (SNP) map of the mouse genome (approximately 8.3 million SNPs) and associated statistical methods allowed us to construct a more generalizable map of the quantitative trait loci (QTL) associated with physical activity. Specifically, we measured wheel running activity in male and female mice (average age 9 wk) in 41 inbred strains and used activity data from 38 of these strains in a haplotype association mapping analysis to determine QTL associated with activity. As seen previously, there was a large range of activity patterns among the strains, with the highest and lowest strains differing significantly in daily distance run (27.4-fold), duration of activity (23.6-fold), and speed (2.9-fold). On a daily basis, female mice ran further (24%), longer (13%), and faster (11%). Twelve QTL were identified, with three (on Chr. 12, 18, and 19) in both male and female mice, five specific to males, and four specific to females. Eight of the 12 QTL, including the 3 general QTL found for both sexes, fell into intergenic areas. The results of this study further support the findings of a moderate to high heritability of physical activity and add general genomic areas applicable to a large number of mouse strains that can be further mined for candidate genes associated with regulation of physical activity. Additionally, results suggest that potential genetic mechanisms arising from traditional noncoding regions of the genome may be involved in regulation of physical activity.

An Epistatic Genetic Basis for Physical Activity Traits in Mice

We recently identified several (4-8) quantitative trait loci (QTL) for 3 physical activity traits (daily distance, duration, and speed voluntarily run) in an F(2) population of mice derived from an original intercross of 2 strains that exhibited large differences in activity. These QTL cumulatively explained from 11% to 34% of the variation in these traits, but this was considerably less than their total genetic variability estimated from differences among inbred strains. We therefore decided to test whether epistatic interactions might account for additional genetic variation in these traits in this same population of mice. We conducted a full genome epistasis scan for all possible interactions of QTL between each pair of 20 chromosomes. The results of this scan revealed an abundance of epistasis, with QTL throughout the genome being involved in significant interactions. Overall, epistatic effects contributed an average of 26% of the total variation among the 3 activity traits. These results suggest that epistatic interactions of genes may play as important a role in the genetic architecture of physical activity traits as single-locus effects and need to be considered in future candidate gene identification studies.

Repeatability of exercise behaviors in mice

PURPOSE Measurements of exercise behaviors in rodents such as maximal treadmill endurance and physical activity are often used in the literature; however, minimal data are available regarding the repeatability of measurements used for these exercise behaviors. This study assessed the repeatability of a commonly used maximal exercise endurance treadmill test as well as voluntary physical activity measured by wheel running in mice. METHODS Repeatability of treadmill tests were analyzed for both inbred and outbred mice in addition to a 10 week repeatability analysis using Balb/cJ mice (n=20). Voluntary daily physical activity was assessed by distance, duration, and speed of wheel running (WR). Physical activity measurements on days 5 and 6 of WR in a large cohort (n=739) of both inbred and outbred mice were compared. RESULTS No significant differences (p>0.05) in exercise endurance were found between different cohorts of Balb/cJ and DBA/2J mice indicating strains overall generally test the same; however, significant differences between tests were seen within BaD2F(2) animals (p<0.001). Bland-Altman analysis revealed a lack of agreement between weekly endurance tests within mouse, and correlation analysis showed lack of consistent correlations between weekly endurance tests within mouse. No significant differences were found for WR measurements within mouse between days (p=0.99). High correlations between days within mouse for WR were found (r=0.74-0.85). CONCLUSIONS High intra-mouse variability between repeated endurance tests suggests that treadmill testing in an enclosed chamber with shock grid for motivation to run in mice is not repeatable. Conversely, high correlation and agreement between days of wheel-running measurements suggest that voluntary activity (WR) is repeatable and stable within individual mice.

The effects of isometric exercise training on resting blood pressure and orthostatic tolerance in humans

Isometric exercise training has been shown to reduce resting blood pressure, but the effect that this might have on orthostatic tolerance is poorly understood. Changes in orthostatic tolerance may also be dependent on whether the upper or lower limbs of the body are trained using isometric exercise. Twenty‐seven subjects were allocated to either a training or control group. A training group first undertook 5 weeks of isometric exercise training of the legs, and after an 8 week intervening period, a second training group containing six subjects from the initial training group, undertook 5 weeks of isometric arm‐training. The control group were asked to continue their normal daily activities throughout the 18 weeks of the study. In all subjects orthostatic tolerance, assessed using lower body negative pressure (LBNP), and resting blood pressure were measured before and after each of the 5 week training or control periods. Estimated lean leg volume was determined before and after leg‐training. During all LBNP tests, heart rate and blood pressure were recorded each minute, and the time taken to reach the highest heart rate was derived (time to peak HR). Resting systolic blood pressure (mean ± s.d.), when measured during the last week of training, was significantly reduced after both leg (‐10 ± 8.7 mmHg) and arm (‐12.4 ± 9.3 mmHg; P < 0.05) isometric exercise training, compared to controls. This reduction disappeared when blood pressure was measured immediately before the LBNP tests, which followed training. Orthostatic tolerance only increased after leg‐training (20.8 ± 16.4 LTI; P < 0.05) and was accompanied by an increased time to peak HR (119.8 ± 106.3 beats min−1; P< 0.05) in this group. Blood pressure responses to LBNP did not change after arm‐training, leg‐training or in controls (P > 0.05). There was a small but significant increase in estimated lean leg volume after leg‐training (0.1 ± 0.1 1; P < 0.05). These results suggest that lower resting blood pressure is probably not responsible for the increased orthostatic tolerance after isometric exercise training of the legs. Rather, it is possible that the training altered some other aspect of cardiovascular control during orthostatic stress that was apparent in the changes in heart rate. Legtraining was accompanied by increases in estimated lean leg volume. The effects of isometric training on orthostatic tolerance appear to be specific to limbs that are directly involved in LBNP testing.

Current Understanding of the Genetic Basis for Physical Activity

Although it is well known that physical activity prevents and ameliorates a large number of conditions and chronic diseases, it is also incontrovertible that physical inactivity is becoming more prevalent. This paradox has led some to suggest that genetic/biological factors influence activity levels as opposed to the classical notion that voluntary activity is solely regulated by environmental factors. There is a plethora of recent data showing that there is considerable genetic influence on activity levels in both humans and animals and emerging evidence suggesting potential genomic locations for those genetic factors. Several independent lines of evidence suggest that dopamine receptor 1 (Drd1) and nescient helix loop helix (Nhlh2) are excellent candidate genes for the regulation of physical activity, with several other potential candidate genes only partially supported. This foundation provides the basis for continuing work to identify additional candidate genes, to identify other genetic factors that are involved in the regulation of physical activity, and to investigate the mechanisms by which these genes and genetic factors regulate activity.

Automated blood pressure measurements during exercise

One of the critical parameters measured during exercise is blood pressure. However, the accurate measurement of systolic and diastolic blood pressure during exercise is difficult with auscultation and impractical with direct arterial techniques. The purpose of this study was to compare an automated system (Colin, Inc. STBP-680) with auscultation in humans during rest and exercise and to compare the automated system with direct arterial blood pressure measurement in a canine model during pharmacological challenges that resulted in a wide range of blood pressure values. Compared with direct arterial blood pressure taken in the canine model, the STBP-680 gave good estimates of diastolic blood pressure and adequately monitored relative changes in systolic blood pressure, diastolic blood pressure, and mean arterial pressure (mean arterial pressures in all instances were calculated as one-third systolic plus two-thirds diastolic blood pressures). Compared with auscultation methods in humans, the STBP-680 gave similar estimates of resting diastolic blood pressure and monitored relative changes in resting systolic blood pressures, diastolic blood pressures, and mean arterial pressures. During both treadmill and cycle ergometer exercise in humans, the STBP-680 monitored changes in systolic blood pressure, phase IV diastolic blood pressure, and mean arterial pressure. Further, the STBP-680 estimated exactly and noted relative changes in heart rate in every test. However, during exercise, quantitative estimations of systolic blood pressure by the STBP-680 were higher than those found using auscultation. Where exact, quantitative measures of blood pressure are needed, direct arterial measurement continues to be the most accurate method. However, where indirect methods can be used, the STBP-680 may provide a suitable alternative that reduces many of the technical concerns of auscultation in young, healthy individuals.

Genetic variation in the pleiotropic association between physical activity and body weight in mice

BackgroundA sedentary lifestyle is often assumed to lead to increases in body weight and potentially obesity and related diseases but in fact little is known about the genetic association between physical activity and body weight. We tested for such an association between body weight and the distance, duration, and speed voluntarily run by 310 mice from the F2 generation produced from an intercross of two inbred lines that differed dramatically in their physical activity levels.MethodsWe used a conventional interval mapping approach with SNP markers to search for QTLs that affected both body weight and activity traits. We also conducted a genome scan to search for relationship QTLs (rel QTLs), or chromosomal regions that affected an activity trait variably depending on the phenotypic value of body weight.ResultsWe uncovered seven quantitative trait loci (QTLs) affecting body weight, but only one co-localized with another QTL previously found for activity traits. We discovered 19 rel QTLs that provided evidence for a genetic (pleiotropic) association of physical activity and body weight. The three genotypes at each of these loci typically exhibited a combination of negative, zero, and positive regressions of the activity traits on body weight, the net effect of which was to produce overall independence of body weight from physical activity. We also demonstrated that the rel QTLs produced these varying associations through differential epistatic interactions with a number of other epistatic QTLs throughout the genome.ConclusionIt was concluded that individuals with specific combinations of genotypes at the rel QTLs and epi QTLs might account for some of the variation typically seen in plots of the association of physical activity with body weight.