Low Running Capacity Research Articles

Enhanced cardiac protein glycosylation (O-GlcNAc) of selected mitochondrial proteins in rats artificially selected for low running capacity

O-linked β-N-acetyl glucosamine (O-GlcNAc) is a posttranslational modification consisting of a single N-acetylglucosamine moiety attached by an O-β-glycosidic linkage to serine and threonine residues of both nuclear and cytosolic proteins. Analogous to phosphorylation, the modification is reversible and dynamic, changing in response to stress, nutrients, hormones, and exercise. Aims of this study were to examine differences in O-GlcNAc protein modification in the cardiac tissue of rats artificially selected for low (LCR) or high (HCR) running capacity. Hyperinsulinemic-euglycemic clamps in conscious animals assessed insulin sensitivity while 2-[(14)C] deoxyglucose tracked both whole body and tissue-specific glucose disposal. Immunoblots of cardiac muscle examined global O-GlcNAc modification, enzymes that control its regulation (OGT, OGA), and specific proteins involved in mitochondrial oxidative phosphorylation. LCR rats were insulin resistant disposing of 65% less glucose than HCR. Global tissue O-GlcNAc, OGT, OGA, and citrate synthase were similar between groups. Analysis of cardiac proteins revealed enhanced O-GlcNAcylation of mitochondrial Complex I, Complex IV, VDAC, and SERCA in LCR compared with HCR. These results are the first to establish an increase in specific protein O-GlcNAcylation in LCR animals that may contribute to progressive mitochondrial dysfunction and the pathogenesis of insulin resistance observed in the LCR phenotype.

Brain Activation Patterns at Exhaustion in Rats That Differ in Inherent Exercise Capacity

In order to further understand the genetic basis for variation in inherent (untrained) exercise capacity, we examined the brains of 32 male rats selectively bred for high or low running capacity (HCR and LCR, respectively). The aim was to characterize the activation patterns of brain regions potentially involved in differences in inherent running capacity between HCR and LCR. Using quantitative in situ hybridization techniques, we measured messenger ribonuclease (mRNA) levels of c-Fos, a marker of neuronal activation, in the brains of HCR and LCR rats after a single bout of acute treadmill running (7.5–15 minutes, 15° slope, 10 m/min) or after treadmill running to exhaustion (15–51 minutes, 15° slope, initial velocity 10 m/min). During verification of trait differences, HCR rats ran six times farther and three times longer prior to exhaustion than LCR rats. Running to exhaustion significantly increased c-Fos mRNA activation of several brain areas in HCR, but LCR failed to show significant elevations of c-Fos mRNA at exhaustion in the majority of areas examined compared to acutely run controls. Results from these studies suggest that there are differences in central c-Fos mRNA expression, and potential brain activation patterns, between HCR and LCR rats during treadmill running to exhaustion and these differences could be involved in the variation in inherent running capacity between lines.

Divergent skeletal muscle respiratory capacities in rats artificially selected for high and low running ability: a role for Nor1?

Inactivity-related diseases are becoming a huge burden on Western society. While there is a major environmental contribution to metabolic health, the intrinsic properties that predispose or protect against particular health traits are harder to define. We used rat models of inborn high running capacity (HCR) and low running capacity (LCR) to determine inherent differences in mitochondrial volume and function, hypothesizing that HCR rats would have greater skeletal muscle respiratory capacity due to an increase in mitochondrial number. Additionally, we sought to determine if there was a link between the expression of the orphan nuclear receptor neuron-derived orphan receptor (Nor)1, a regulator of oxidative metabolism, and inherent skeletal muscle respiratory capacity. LCR rats were 28% heavier (P < 0.0001), and fasting serum insulin concentrations were 62% greater than in HCR rats (P = 0.02). In contrast, HCR rats had better glucose tolerance and reduced adiposity. In the primarily oxidative soleus muscle, maximal respiratory capacity was 21% greater in HCR rats (P = 0.001), for which the relative contribution of fat oxidation was 20% higher than in LCR rats (P = 0.02). This was associated with increased citrate synthase (CS; 33%, P = 0.009) and β-hydroxyacyl-CoA (β-HAD; 33%, P = 0.0003) activities. In the primarily glycolytic extensor digitum longus muscle, CS activity was 29% greater (P = 0.01) and β-HAD activity was 41% (P = 0.0004) greater in HCR rats compared with LCR rats. Mitochondrial DNA copy numbers were also elevated in the extensor digitum longus muscles of HCR rats (35%, P = 0.049) and in soleus muscles (44%, P = 0.16). Additionally, HCR rats had increased protein expression of individual mitochondrial respiratory complexes, CS, and uncoupling protein 3 in both muscle types (all P < 0.05). In both muscles, Nor1 protein was greater in HCR rats compared with LCR rats (P < 0.05). We propose that the differential expression of Nor1 may contribute to the differences in metabolic regulation between LCR and HCR phenotypes.

Susceptibility to intracerebral hemorrhage-induced brain injury segregates with low aerobic capacity in rats

Although low exercise capacity is a risk factor for stroke, the exact mechanisms that underlie this connection are not known. As a model system for exploring the association between aerobic capacity and disease risks we applied two-way artificial selection over numerous generations in rats to produce low capacity runners (LCR) and high capacity runners (HCR). Here we compared intracerebral hemorrhage (ICH)-induced brain injury in both genders of these rat lines. HCR and LCR rats had 100μl blood injected into the right caudate and were killed at days 1, 3, 7 and 28 for brain water content determination, immunohistochemistry, histology, Western blot, and behavioral tests. Compared to male HCRs, male LCRs had more severe ICH-induced brain injury including worse brain edema, necroptosis, brain atrophy, and neurological deficits, but not increased numbers of Fluoro-Jade C positive cells or elevated cleaved caspase-3 levels. This was associated with greater microglial activation, and heme oxygenase-1 and protease activated receptor (PAR)-1 upregulation. In females, edema was also greater in LCRs than in HCRs, although it was less severe in females than in males for both LCRs and HCRs. Thus, ICH-induced brain injury was more severe in LCRs, a model of low exercise capacity, than in HCRs. Increased activation of microglia and PAR-1 may participate mechanistically in increased ICH-susceptibility. Females were protected against ICH-induced brain edema formation in both HCRs and LCRs.

Artificial Selection for High Aerobic Capacity is Protective against Weight Gain via Thermogenesis?

Higher spontaneous activity and non‐exercise activity thermogenesis (NEAT) contribute significantly to daily energy balance, body weight, and therefore in health. It was previously shown that, relative to body mass, rats selectively bred for high intrinsic aerobic capacity (HCR = high‐capacity runner) consume more energy and are more spontaneously active than low‐capacity runner (LCR) counterparts. We tested whether HCRs have higher rectal temperatures after 2 h fasting and during glucose tolerance test compared to LCRs. We studied 40 HCR/LCR female rats for the fasting condition, and 20 HCR/LCR female rats for the glucose tolerance test. Rats were from 27th generation of selection and aged 9 months. During the glucose tolerance test we controlled the spontaneous activity levels of the rats with ground reaction force recording. Our study revealed that after 2 h fasting HCRs had significantly higher rectal temperature compared to LCRs (p &lt; 0.05). During glucose tolerance test the rectal temperature of HCRs was significantly higher compared to LCRs throughout the test protocol (p &lt; 0.05). In the present study the spontaneous activity of HCRs was higher at the beginning (p &lt; 0.05), but not at the end of the glucose tolerance test, suggesting that higher activity of HCR rats does not exclusively explain higher NEAT. This study was supported by the Ministry of Education (Finland) and the National Institute of Health.

Targeted protein glycosylation (O‐GlcNAc) of mitochondrial proteins in rats selected for low running capacity

Excess flux through the hexosamine biosynthesis pathway results in enhanced protein modification by O‐linked‐β‐N‐acetylglucosamine (O‐GlcNAc), glucotoxicity and insulin resistance. Contributing factors are complex and may include environment as well as genetics. Aims were to examine differences in O‐GlcNAc in rats artificially selected for either low (LCR) or high (HCR) running capacity. Reduced fitness is known to impair mitochondrial function, perpetuate O‐GlcNAc modification and contribute to reduced insulin sensitivity. Insulin sensitivity was assessed by a hyperinsulinemic‐euglycemic clamp in conscious animals while 2‐[14C]deoxyglucose tracked glucose disposal. Immunoblots of heart tissue examined O‐GlcNAc levels and enzymes that control its regulation (OGT, OGA). Artificial selection of rats to generation 19 resulted in 6‐fold greater running capacity and 63% reduction in fat mass in HCR compared to LCR (p&lt;0.05). LCR rats were insulin resistant disposing of 65% less glucose than HCR (p&lt;0.05). Tissue analysis revealed enhanced O‐GlcNAcylation of Complex I, Complex IV, VDAC and SERCA in LCR compared to HCR (p&lt;0.05). Levels of OGT and OGA were not different between groups. Thus, increased O‐GlcNAc in LCR animals may contribute to mitochondrial dysfunction and the pathogenesis of insulin resistance.

Overweight female rats selectively breed for low aerobic capacity exhibit increased myocardial fibrosis and diastolic dysfunction

The statistical association between endurance exercise capacity and cardiovascular disease suggests that impaired aerobic metabolism underlies the cardiovascular disease risk in men and women. To explore this connection, we applied divergent artificial selection in rats to develop low-capacity runner (LCR) and high-capacity runner (HCR) rats and found that disease risks segregated strongly with low running capacity. Here, we tested if inborn low aerobic capacity promotes differential sex-related cardiovascular effects. Compared with HCR males (HCR-M), LCR males (LCR-M) were overweight by 34% and had heavier retroperitoneal, epididymal, and omental fat pads; LCR females (LCR-F) were 20% heavier than HCR females (HCR-F), and their retroperitoneal, but not perireproductive or omental, fat pads were heavier as well. Unlike HCR-M, blood pressure was elevated in LCR-M, and this was accompanied by left ventricular (LV) hypertrophy. Like HCR-F, LCR-F exhibited normal blood pressure and LV weight as well as increased spontaneous cage activity compared with males. Despite normal blood pressures, LCR-F exhibited increased myocardial interstitial fibrosis and diastolic dysfunction, as indicated by increased LV stiffness, a decrease in the initial filling rate, and an increase in diastolic relaxation time. Although females exhibited increased arterial stiffness, ejection fraction was normal. Increased interstitial fibrosis and diastolic dysfunction in LCR-F was accompanied by the lowest protein levels of phosphorylated AMP-actived protein kinase [phospho-AMPK (Thr(172))] and silent information regulator 1. Thus, the combination of risk factors, including female sex, intrinsic low aerobic capacity, and overweightness, promote myocardial stiffness/fibrosis sufficient to induce diastolic dysfunction in the absence of hypertension and LV hypertrophy.

Selective breeding for endurance running capacity affects cognitive but not motor learning in rats

The ability to utilize oxygen has been shown to affect a wide variety of physiological factors often considered beneficial for survival. As the ability to learn can be seen as one of the core factors of survival in mammals, we studied whether selective breeding for endurance running, an indication of aerobic capacity, also has an effect on learning. Rats selectively bred over 23 generations for their ability to perform forced treadmill running were trained in an appetitively motivated discrimination-reversal classical conditioning task, an alternating T-maze task followed by a rule change (from a shift-win to stay-win rule) and motor learning task. In the discrimination-reversal and T-maze tasks, the high-capacity runner (HCR) rats outperformed the low-capacity runner (LCR) rats, most notably in the phases requiring flexible cognition. In the Rotarod (motor-learning) task, the HCR animals were overall more agile but learned at a similar rate with the LCR group as a function of training. We conclude that the intrinsic ability to utilize oxygen is associated especially with tasks requiring plasticity of the brain structures implicated in flexible cognition.

Determination of Minimum Alveolar Concentration for Isoflurane and Sevoflurane in a Rodent Model of Human Metabolic Syndrome

Morbid obesity affects the pharmacokinetics and pharmacodynamics of anesthetics, which may result in inappropriate dosing. We hypothesized that obesity significantly alters the minimum alveolar concentration (MAC) for isoflurane and sevoflurane. To test this hypothesis, we used a rodent model of human metabolic syndrome developed through artificial selection for inherent low aerobic capacity runners (LCR) and high aerobic capacity runners (HCR). The LCR rats are obese, display phenotypes homologous to those characteristic of human metabolic syndrome, and exhibit low running endurance. In contrast, HCR rats have high running endurance and are characterized by improved cardiovascular performance and overall health. Male and female LCR (n = 10) and HCR (n = 10) rats were endotracheally intubated and maintained on mechanical ventilation with either isoflurane or sevoflurane. A bracketing design was used to determine MAC; sensory stimulation was induced by tail clamping. An equilibration period of 30 minutes was provided before and between the consecutive tail clamps. Two-tailed parametric (unpaired t test) and nonparametric (Mann-Whitney test) statistics were used for the comparison of MAC between LCR and HCR rats. The data are reported as mean ± sd along with the 95% confidence interval. A P value of <0.05 was considered statistically significant. The MAC for isoflurane in LCR rats (1.52% ± 0.13%) was similar to previously reported isoflurane-MAC for normal rats (1.51% ± 0.12%). The HCR rats showed a significantly higher isoflurane-MAC (1.90% ± 0.19%) than did the LCR rats (1.52% ± 0.13%) (P = 0.0001). The MAC for sevoflurane was not significantly different between LCR and HCR rats and was similar to the previously published sevoflurane-MAC for normal rats (2.4% ± 0.30%). There was no influence of sex on the MAC of either isoflurane or sevoflurane. Obesity and associated comorbidities do not affect anesthetic requirements as measured by MAC in a rodent model of metabolic syndrome. By contrast, high aerobic capacity is associated with a higher MAC for isoflurane and may be a risk factor for subtherapeutic dosing.

In This Issue

HomeCirculation ResearchVol. 109, No. 10In This Issue Free AccessIn BriefPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessIn BriefPDF/EPUBIn This Issue Originally published28 Oct 2011https://doi.org/10.1161/RES.0b013e31823affe5Circulation Research. 2011;109:1097MicroRNA-29 in Aortic Aneurysm Formation (p 1115)Boon et al find that blocking a particular microRNA prevents aortic aneurysm.As we age, our blood vessels become weaker and more prone to aneurysms. Indeed, abdominal aortic aneurysm is a problem that affects approximately 9% of elderly men. Growth and development of normal blood vessel is regulated by, among other things, microRNAs (miRs), so Boon et al wondered whether miRs might also play a role in vessel aging and pathology. The team examined the aortas of old and young mice and found that 18 miRs were differentially expressed. However, only one of these, miR-29, controlled mRNA expression. miR-29 was upregulated not only in aging aortas, but also in mouse models of aortic aneurysm and in human aneurysm biopsies. miR-29 had previously been shown to repress the expression of extracellular matrix proteins in the heart after infarction, thereby reducing fibrosis and scarring. These same ECM mRNAs were the targets for miR-29 in aging aortas, but in this case the team showed that the result was vessel weakening. Inhibiting miR-29 prevented the downregulation of ECM proteins and, more importantly, the formation of aneurysms in aged mice. Localized inhibition of miR-29 at the site of an aortic aneurysm might therefore be a way to aid repair, strengthen the vessel wall, and prevent future aneurysms.Download figureDownload PowerPointHypoxia and Macrophage Lipid Content (p 1141)Mice are good models for studying hypoxia in atherosclerosis, say Parathath et al.The cores of human atherosclerotic plaques are often far enough from the passing blood flow that they are deprived of an oxygen supply. This hypoxic environment is associated with the activation of a transcription factor called hypoxia inducible factor (HIF-1), which regulates genes involved in apoptosis, metabolism, inflammation, and other processes relevant to atherosclerosis. Whether hypoxia and HIF-1 are actually involved in atherogenesis, however, is not known. Mouse models of atherosclerosis were considered irrelevant for studying the role of hypoxia in atherogenesis because it was thought that due to the small size of mouse plaques oxygen could still reach the cores. Parathath et al now show that, in fact, mouse plaques do express HIF-1 as well as its transcriptional targets. The team also showed that mouse macrophages had altered lipid metabolism under hypoxic conditions, with increased sterol content and decreased cholesterol efflux – changes that would likely worsen atherosclerosis. Blocking the activity of HIF-1 prevented these hypoxia effects, indicating the transcription factor was directly responsible. The findings of Parathath et al suggest that hypoxia has a negative impact on atherogenesis and that mice would be useful for studying the damaging effect of hypoxia and for finding ways to combat it.Download figureDownload PowerPointLinking Exercise Capacity With Mortality (p 1162)A genetic proclivity for exercise is linked to longevity, report Koch et al.That exercise decreases the risk for cardiovascular disease is well-known, but Koch et al now show that you can be born with this benefit. Well, at least rats can. The researchers selectively bred rats over 14 or so generations until they had populations of low capacity runners (LCRs) and high capacity runners (HCRs), as determined by treadmill testing. Then, they compared oxygen uptake, myocardial function, endurance performance, and body mass in adult and aged rats from the 2 groups. As the rats progressed from adulthood to old age, the HCRs had sustained activity levels, energy expenditure, and lean body mass. They also had lower blood pressure, improved cardiomyocyte function – assessed by contractility, morphology and intracellular calcium handling – and longer lifespans. The finding that longevity is linked with a genetic propensity for exercise does not mean that those of us who are naturally less active will not reap the same benefits from exercise. Rather, the research suggests that the LCR and HCR model rats will be excellent tools for studying the biological pathways that link aerobic activity with healthy aging and longevity – work likely to benefit everybody.Download figureDownload PowerPointWritten by Ruth Williams Previous Back to top Next FiguresReferencesRelatedDetails October 28, 2011Vol 109, Issue 10 Advertisement Article InformationMetrics © 2011 American Heart Association, Inc.https://doi.org/10.1161/RES.0b013e31823affe5 Originally publishedOctober 28, 2011 PDF download Advertisement

Changes in mRNA levels for brain-derived neurotrophic factor after wheel running in rats selectively bred for high- and low-aerobic capacity

We evaluated levels of exercise-induced brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) within the hippocampal formation in rats selectively bred for 1) high intrinsic (i.e., untrained) aerobic capacity (High Capacity Runners, HCR), 2) low intrinsic aerobic capacity (Low Capacity Runners, LCR), and 3) unselected Sprague–Dawley (SD) rats with or without free access to running wheels for 3weeks. The specific aim of the study was to determine whether a dose–response relationship exists between cumulative running distance and levels of BDNF mRNA. No additional treatments or behavioral manipulations were used. HCR, LCR, and SD rats were grouped by strain and randomly assigned to sedentary or activity (voluntary access to activity wheel) conditions. Animals were killed after 21days of exposure to the assigned conditions. Daily running distances (mean±standard deviation meters/day) during week three were: HCR (4726±3220), SD (2293±3461), LCR (672±323). Regardless of strain, levels of BDNF mRNA in CA1 were elevated in wheel runners compared to sedentary rats and this difference persisted after adjustment for age (p=0.040). BDNF mRNA was not affected by intrinsic aerobic capacity and was not related to total running distance. The results support that BDNF mRNA expression is increased by unlimited access to activity wheel running for 3weeks but is not dependent upon accumulated running distance.

Proteomic analysis reveals perturbed energy metabolism and elevated oxidative stress in hearts of rats with inborn low aerobic capacity

Selection on running capacity has created rat phenotypes of high-capacity runners (HCRs) that have enhanced cardiac function and low-capacity runners (LCRs) that exhibit risk factors of metabolic syndrome. We analysed hearts of HCRs and LCRs from generation 22 of selection using DIGE and identified proteins from MS database searches. The running capacity of HCRs was six-fold greater than LCRs. DIGE resolved 957 spots and proteins were unambiguously identified in 369 spots. Protein expression profiling detected 67 statistically significant (p<0.05; false discovery rate <10%, calculated using q-values) differences between HCRs and LCRs. Hearts of HCR rats exhibited robust increases in the abundance of each enzyme of the β-oxidation pathway. In contrast, LCR hearts were characterised by the modulation of enzymes associated with ketone body or amino acid metabolism. LCRs also exhibited enhanced expression of antioxidant enzymes such as catalase and greater phosphorylation of α B-crystallin at serine 59, which is a common point of convergence in cardiac stress signalling. Thus, proteomic analysis revealed selection on low running capacity is associated with perturbations in cardiac energy metabolism and provided the first evidence that the LCR cardiac proteome is exposed to greater oxidative stress.

Low intrinsic running capacity is associated with reduced skeletal muscle substrate oxidation and lower mitochondrial content in white skeletal muscle

Chronic metabolic diseases develop from the complex interaction of environmental and genetic factors, although the extent to which each contributes to these disorders is unknown. Here, we test the hypothesis that artificial selection for low intrinsic aerobic running capacity is associated with reduced skeletal muscle metabolism and impaired metabolic health. Rat models for low- (LCR) and high- (HCR) intrinsic running capacity were derived from genetically heterogeneous N:NIH stock for 20 generations. Artificial selection produced a 530% difference in running capacity between LCR/HCR, which was associated with significant functional differences in glucose and lipid handling by skeletal muscle, as assessed by hindlimb perfusion. LCR had reduced rates of skeletal muscle glucose uptake (∼30%; P = 0.04), glucose oxidation (∼50%; P = 0.04), and lipid oxidation (∼40%; P = 0.02). Artificial selection for low aerobic capacity was also linked with reduced molecular signaling, decreased muscle glycogen, and triglyceride storage, and a lower mitochondrial content in skeletal muscle, with the most profound changes to these parameters evident in white rather than red muscle. We show that a low intrinsic aerobic running capacity confers reduced insulin sensitivity in skeletal muscle and is associated with impaired markers of metabolic health compared with high intrinsic running capacity. Furthermore, selection for high running capacity, in the absence of exercise training, endows increased skeletal muscle insulin sensitivity and oxidative capacity in specifically white muscle rather than red muscle. These data provide evidence that differences in white muscle may have a role in the divergent aerobic capacity observed in this generation of LCR/HCR.

Osteoblast Response to Ovariectomy Is Enhanced in Intrinsically High Aerobic-Capacity Rats

The role of exercise in promoting bone health is typically attributed to increased mechanical loading, which induces functional adaptation. Recent evidence suggests that habitual aerobic exercise has influence at the cellular level as well. The effect of aerobic capacity on osteoblast-lineage cell differentiation and function as well as skeletal phenotype is unknown. Using a rat model of high-capacity and low-capacity runners (HCRs and LCRs, respectively), in which an intrinsic functional genomic difference in aerobic capacity exists between nontrained animals, this study evaluated the effects of aerobic capacity on measures of bone mass and strength as well as osteoblast activity following ovariectomy. The ovariectomized rat emulates the clinical features of the estrogen-depleted human skeleton and represents a valuable model for studying short-term upregulation of osteoblast activity. We hypothesized that intrinsically high aerobic capacity would augment osteoblast response, which would mitigate the deleterious effects of hormone withdrawal. Femora and tibiae were assessed by micro-computed tomography, mechanical testing, and dynamic histomorphometry. HCRs had enhanced femoral tissue mineral density and estimated elastic modulus relative to LCRs. At 4 weeks postovariectomy, HCRs demonstrated a more robust osteoblast response. Markers of bone formation were upregulated to a greater extent in HCRs than LCRs, suggesting a role for aerobic capacity in governing osteoblast activity. Results from this and future studies will help to identify the influence of cellular aerobic metabolism on bone health, which may lead to new strategies for targeting diseases of the skeleton.

Lower oxidative DNA damage despite greater ROS production in muscles from rats selectively bred for high running capacity

Artificial selection in rat has yielded high-capacity runners (HCR) and low-capacity runners (LCR) that differ in intrinsic (untrained) aerobic exercise ability and metabolic disease risk. To gain insight into how oxygen metabolism may have been affected by selection, we compared mitochondrial function, oxidative DNA damage (8-dihydroxy-guanosine; 8dOHG), and antioxidant enzyme activities in soleus muscle (Sol) and gastrocnemius muscle (Gas) of adult and aged LCR vs. HCR rats. In Sol of adult HCR rats, maximal ADP-stimulated respiration was 37% greater, whereas in Gas of adult HCR rats, there was a 23% greater complex IV-driven respiratory capacity and 54% greater leak as a fraction of electron transport capacity (suggesting looser mitochondrial coupling) vs. LCR rats. H(2)O(2) emission per gram of muscle was 24-26% greater for both muscles in adult HCR rats vs. LCR, although H(2)O(2) emission in Gas was 17% lower in HCR, after normalizing for citrate synthase activity (marker of mitochondrial content). Despite greater H(2)O(2) emission, 8dOHG levels were 62-78% lower in HCR rats due to 62-96% higher superoxide dismutase activity in both muscles and 47% higher catalase activity in Sol muscle in adult HCR rats, with no evidence for higher 8 oxoguanine glycosylase (OGG1; DNA repair enzyme) protein expression. We conclude that genetic segregation for high running capacity has generated a molecular network of cellular adaptations, facilitating a superior response to oxidative stress.

Exercise training reverses impaired skeletal muscle metabolism induced by artificial selection for low aerobic capacity

We have used a novel model of genetically imparted endurance exercise capacity and metabolic health to study the genetic and environmental contributions to skeletal muscle glucose and lipid metabolism. We hypothesized that metabolic abnormalities associated with low intrinsic running capacity would be ameliorated by exercise training. Selective breeding for 22 generations resulted in rat models with a fivefold difference in intrinsic aerobic capacity. Low (LCR)- and high (HCR)-capacity runners remained sedentary (SED) or underwent 6 wk of exercise training (EXT). Insulin-stimulated glucose transport, insulin signal transduction, and rates of palmitate oxidation were lower in LCR SED vs. HCR SED (P < 0.05). Decreases in glucose and lipid metabolism were associated with decreased β₂-adrenergic receptor (β₂-AR), and reduced expression of Nur77 target proteins that are critical regulators of muscle glucose and lipid metabolism [uncoupling protein-3 (UCP3), fatty acid transporter (FAT)/CD36; P < 0.01 and P < 0.05, respectively]. EXT reversed the impairments to glucose and lipid metabolism observed in the skeletal muscle of LCR, while increasing the expression of β₂-AR, Nur77, GLUT4, UCP3, and FAT/CD36 (P < 0.05) in this tissue. However, no metabolic improvements were observed following exercise training in HCR. Our results demonstrate that metabolic impairments resulting from genetic factors (low intrinsic aerobic capacity) can be overcome by an environmental intervention (exercise training). Furthermore, we identify Nur77 as a potential mechanism for improved skeletal muscle metabolism in response to EXT.

Risk-assessment and Coping Strategies Segregate with Divergent Intrinsic Aerobic Capacity in Rats

Metabolic function is integrally related to an individual's susceptibility to, and progression of, disease. Selective breeding for intrinsic treadmill running in rats has produced distinct lines of high- or low-capacity runners (HCR and LCR, respectively) that exhibit numerous physiological differences. To date, the role of intrinsic aerobic capacity on behavior and stress response in these rats has not been addressed and was the focus of these studies. HCR and LCR rats did not differ in their locomotor response to novelty or behavior in the light/dark box. In contrast, immobility in the forced swim test was higher in LCR rats compared with HCR rats, regardless of desipramine treatment. Although both HCR and LCR rats responded to cat odor with decreased exploration and increased risk assessment, HCR rats showed greater contextual conditioning to cat odor. HCR rats exhibited higher expression of corticotropin-releasing hormone in the central nucleus of the amygdala, as well as heavier adrenal and thymus weight. Corticosterone was comparable among HCR and LCR rats at light/dark transitions, and in response to unavoidable cat odor. HCR rats, however, exhibited a greater corticosterone response following the light/dark box. These experiments show that the LCR phenotype associates with decreased risk assessment in response to salient danger signals and passive coping. In contrast, HCR rats show a more naturalistic strategy in that they employ active coping and a more vigilant and cautious response to environmental novelty and salient danger signals. Within this context, we propose that intrinsic aerobic capacity is a central feature mechanistically linking complex metabolic disease and behavior.

Rats Bred For Low Aerobic Capacity Become Rapidly Fatigued And Have Slow Metabolic Recovery After Stimulated Muscle Contractions

Good aerobic capacity has a hypothesized role in increased health and longevity. Since impaired oxidative metabolism underlies several complex diseases, such as type 2 diabetes and cardiovascular disease, it is generally thought that the ability to increase oxygen transport and utilization capacity by exercise training stimulus helps prevent one from developing the aforementioned diseases. However, the magnitude of exercise capacity differs substantially between individuals. Primary assumption is that this heterogeneity is the consequence of variation distributed across a large number of genes for both the intrinsic (untrained) aerobic capacity and that accrued from exercise training. To address the intrinsic component, Koch and Britton (Physiol Genomics, 2001) developed two rat strains by artificial selection for low and high endurance treadmill running capacity in the untrained condition. These contrasting rat models, termed low capacity runner (LCR) and high capacity runner (HCR), are excellent biological models for evaluating the interplay of genetic and environmental factors as determinants of high health and aerobic capacity. PURPOSE: Our long-term aim is to evaluate the influence of life-long aerobic exercise training on aging and longevity using the LCR/HCR model system. Such studies require the application of non-invasive functional measures across time. Here we report on our first measurements of skeletal muscle energy metabolism in non-trained LCR and HCR rats. METHODS: Muscle function was studied in electrically stimulated contracting gastrocnemius muscle using 1H-magnetic resonance imaging (MRI) and 31P-MR spectroscopy. The simultaneous force output was measured with an ergometer connected to a foot pedal. RESULTS: Our results from mechanical measurement (force output) show a significant difference between LCR and HCR groups for the decrease in the isometric force during transcutaneous stimulation, the decrease being greater in LCR rats (P<0,001). 31P-MR spectroscopy results demonstrated that phosphocreatine resynthesis after stimulation was significantly slower in LCR group (P<0,02). CONCLUSIONS: Consistent with diminished treadmill running capacity, this study shows that skeletal muscle in LCR rats becomes fast fatigued during maximal muscle stimulation. 31P-MR spectroscopy seems to be a valid non-invasive method to investigate consecutive differentials in skeletal muscle energy metabolism between the LCR and HCR rats, making longitudinal studies feasible. Supported by the Ministry of Education and Culture (Finland) and the National Center for Research Resources (a component of the NIH).

Caloric Restriction Reverses Hepatic Insulin Resistance and Steatosis in Rats with Low Aerobic Capacity

Rats selectively bred for low aerobic running capacity exhibit the metabolic syndrome, including hyperinsulinemia, insulin resistance, visceral obesity, and dyslipidemia. They also exhibit features of nonalcoholic steatohepatitis, including chicken-wire fibrosis, inflammation, and oxidative stress. Hyperinsulinemia in these rats is associated with impaired hepatic insulin clearance. The current studies aimed to determine whether these metabolic abnormalities could be reversed by caloric restriction (CR). CR by 30% over a period of 2-3 months improved insulin clearance in parallel to inducing the protein content and activation of the carcinoembryonic antigen-related cell adhesion molecule 1, a main player in hepatic insulin extraction. It also reduced glucose and insulin intolerance and serum and tissue (liver and muscle) triglyceride levels. Additionally, CR reversed inflammation, oxidative stress, and fibrosis in liver. The data support a significant role of CR in the normalization of insulin and lipid metabolism in liver.

Locus coeruleus galanin expression is enhanced after exercise in rats selectively bred for high capacity for aerobic activity

The neuropeptide galanin extensively coexists with norepinephrine in locus coeruleus (LC) neurons. Previous research in this laboratory has demonstrated that unlimited access to activity wheels in the home cage increases mRNA for galanin (GAL) in the LC, and that GAL mediates some of the beneficial effects of exercise on brain function. To assess whether capacity for aerobic exercise modulates this upregulation in galanin mRNA, three heterogeneous rat models were tested: rats selectively bred for (1) high intrinsic (untrained) aerobic capacity (High Capacity Runners, HCR) and (2) low intrinsic aerobic capacity (Low Capacity Runners, LCR) and (3) unselected Sprague–Dawley (SD) rats with and without free access to running wheels for 3 weeks. Following this exercise protocol, mRNA for tyrosine hydroxylase (TH) and GAL was measured in the LC. The wheel running distances between the three models were significantly different, and age contributed as a significant covariate. Both selection and wheel access condition significantly affected GAL mRNA expression, but not TH mRNA expression. GAL was elevated in exercising HCR and SD rats compared to sedentary rats while LCR rats did not differ between conditions. Overall running distance significantly correlated with GAL mRNA expression, but not with TH mRNA expression. No strain differences in GAL or TH gene expression were observed in sedentary rats. Thus, intrinsic aerobic running capacity influences GAL gene expression in the LC only insofar as actual running behavior is concerned; aerobic capacity does not influence GAL expression in addition to changes associated with running.