The association of ACTN3 R577X polymorphism with sports specificity in Japanese elite athletes.

The α-actinin-3 proteins regulate muscle function and are located in the Z-line of the fast skeletal muscle. A common null polymorphism of R577X in α-actinin-3 gene (ACTN3) results in its complete absence in fast-twitch muscles. The ACTN3 R577X polymorphism is associated with sprint/power performance in athletes. However, little is known about how this polymorphism impacts sports other than sprint/power-oriented sports in Japanese elite athletes. The aim of our study was to examine the association between ACTN3 R577X polymorphism and elite athlete status in various sports categorized as power/sprint, endurance, artistic, martial arts, and ball game sports. The subjects included 906 Japanese elite athletes and 649 Japanese controls. We analysed the genotype frequency of the ACTN3 R577X polymorphism in sprint/power (n = 120), endurance (n = 150), artistic (n = 45), martial arts (n = 94), and ball game (n = 497) sports athletes. A higher number of sprint/power athletes were R allele carriers compared to the controls, and the endurance and artistic athletes (OR = 1.69, 1.83, and 2.36, 95% CI: 1.02-2.79, 1.02-3.31, and 1.08-5.13, respectively). The frequency of RR genotype was higher in sprint/power, martial arts, and ball game sports athletes (OR = 1.61, 1.84, and 1.39, 95% CI: 1.04-2.50, 1.11-2.95, and 1.05-1.83, respectively) compared to control. Furthermore, there is a significant linear trend with increasing R allele according to athletic status (P for trend < 0.05). The ACTN3 R allele is positively associated with sports performance requiring explosive power such as sprint/power, martial arts, and ball game sports categories.

Comprehensive analysis of common and rare mitochondrial DNA variants in elite Japanese athletes: a case-control study.

The purpose of the present study was to identify mitochondrial DNA (mtDNA) polymorphisms and rare variants that associate with elite Japanese athletic status. Subjects comprised 185 elite Japanese athletes who had represented Japan at international competitions (that is, 100 endurance/middle-power athletes: EMA; 85 sprint/power athletes: SPA) and 672 Japanese controls (CON). The entire mtDNA sequences (16 569 bp) were analyzed by direct sequencing. Nucleotide variants were detected at 1488 sites in the 857 entire mtDNA sequences. A total of 311 variants were polymorphisms (minor allele frequency 1% in CON), and the frequencies of these polymorphisms were compared among the three groups. The EMA displayed excess of seven polymorphisms, including subhaplogroup D4e2- and D4g-specific polymorphisms, compared with CON (P<0.05), whereas SPA displayed excess of three polymorphisms and dearth of nine polymorphisms, including haplogroup G- and subhaplogroup G2a-specific polymorphisms, compared with CON (P<0.05). The frequencies of 10 polymorphisms, including haplogroup G- and subhaplogroup G2a-specific polymorphisms, were different between EMA and SPA (P<0.05): although none of these polymorphisms differed significantly between groups after correcting for multiple comparison (false discovery rate q-value 0.05). The number of rare variants in the 12S ribosomal RNA and NADH dehydrogenase subunit I genes were also higher in SPA than in CON (P<0.05). Analysis of the entire mtDNA of elite Japanese athletes revealed several haplogroup- and subhaplogroup-specific polymorphisms to be potentially associated with elite Japanese athletic status.

Mitochondrial haplogroups associated with elite Japanese athlete status.

PURPOSE: It has been hypothesised that certain mitochondrial haplogroups, which are defined by the presence of a characteristic cluster of tightly linked mitochondrial DNA polymorphisms, would be associated with elite Japanese athlete status. To examine this hypothesis, the frequencies of mitochondrial haplogroups found in elite Japanese athletes were compared with those in the general Japanese population. METHODS: Subjects comprised 139 Olympic athletes (79 endurance/middle-power athletes (EMA), 60 sprint/power athletes (SPA)) and 672 controls (CON). Two mitochondrial DNA fragments containing the hypervariable sequence I (m16024-m16383) of the major non-coding region and the polymorphic site at m.5178C>A within the NADH dehydrogenase subunit 2 gene were sequenced, and subjects were classified into 12 major mitochondrial haplogroups (ie, F, B, A, N9a, N9b, M7a, M7b, M*, G2, G1, D5 or D4). The mitochondrial haplogroup frequency differences among EMA, SPA and CON were then examined. RESULTS: EMA showed an excess of haplogroup G1 (OR 2.52, 95% CI 1.05 to 6.02, p=0.032), with 8.9% compared with 3.7% in CON, whereas SPA displayed a greater proportion of haplogroup F (OR 2.79, 95% CI 1.28 to 6.07, p=0.007), with 15.0% compared with 6.0% in CON. CONCLUSIONS: The results suggest that mitochondrial haplogroups G1 and F are associated with elite EMA and SPA status in Japanese athletes, respectively.

Morning Heart Rate Variability as an Indication of Fatigue Status in Badminton Players during a Training Camp.

This study aimed to clarify whether changes in the fatigue status of elite athletes during a precompetition period could be evaluated using morning heart rate variability (HRV) indices. Eight Japanese National Badminton Team players (age, 23.0 ± 2.8 years) participated in this study. HRV and subjective fatigue were measured during the first (days 1-4: Phase 1) and the second half (days 5-8: Phase 2) of an 8-day national team training camp. The global and parasympathetic HRV indices were as follows: standard deviation of all R-R intervals (SDNN) (Phase 1, 87.5 ms; Phase 2, 104.3 ms; p < 0.05), root mean square of the successive R-R interval differences (RMSSD) (Phase 1, 66.6 ms; Phase 2, 103.6 ms; p < 0.05), and high-frequency component power (HF) (Phase 1, 1412.0 ms2; Phase 2, 3318.5 ms2; p < 0.05). All the aforementioned indices increased significantly from Phase 1 to Phase 2. Significant correlations were observed between the change in subjective fatigue and changes in SDNN, RMSSD, and HF (ρ = -0.80, p = 0.017; ρ = -0.77, p = 0.027; and ρ = -0.80, p = 0.017, respectively). Measuring morning HRV indices may be effective for objectively evaluating changes in the fatigue status of elite athletes during a precompetition period.

Extract from Acanthopanax senticosus harms (Siberian ginseng) activates NTS and SON/PVN in the rat brain.

The extract of the stem bark of Siberian ginseng, Acanthopanax senticosus Harms (ASH), is believed to play a body-coping role in stress through a brain noradrenergic mechanism. The present study was carried out to investigate the effect of ASH on the neuronal activation patterns of c-Fos expression in the rat brain. With ASH administration, c-Fos accumulated in both the supraoptic nuclei (SON) and paraventricular nuclei (PVN), which regulate stress response. Only the caudal regions in the nucleus of the solitary tract (NTS), a locus innervating both the SON and PVN, were activated. Such a neuro-anatomical pattern associated with ASH suggests the possible involvement of these stress-related brain loci.

Inhibitory effects of an orexin-2 receptor antagonist on orexin A- and stress-induced ACTH responses in conscious rats.

Orexins, recognized for their diverse functions in sleep/wakefulness/arousal and appetite regulation, may play provocative roles in stress response. Although the PVN of the hypothalamus expresses an abundance of orexin-2 receptor (OX-2R), the involvement of OX-2R in regulating ACTH response to stress remains unclear. To address this, we examined effects of a selective antagonist to OX-2R (N-{(1S)-1-[6,7-dimethoxy-3,4-dihydro-2(1H)-isoquinolinyl]carbonyl}-2,2-dimethylpropyl)-N-{4-pyridinylmethyl}amine upon plasma ACTH concentrations after administration of orexin A and swimming stress. Increases in ACTH levels with orexin A or swimming stress were attenuated with prior administration of an OX-2R antagonist. These results suggest that swimming stress facilitates ACTH release, at least in part via activation of OX-2R.

Threshold-like pattern of neuronal activation in the hypothalamus during treadmill running: establishment of a minimum running stress (MRS) rat model.

Despite the indication that the hypothalamo-pituitary-adrenal (HPA) axis is activated during treadmill running, there have not been any studies focusing on the relationship between exercise intensity and region-specific neural activities in hypothalamus. To address this, rats were subjected to 30 min of running, either at middle (supra-LT, 25 m min(-1)) or low speeds (sub-LT, 15 m min(-1)), and c-Fos-(+) cells were counted and compared with control rats. Significant increases in blood glucose and lactate levels, and plasma ACTH and osmolality levels were observed during supra-LT running. Only supra-LT running significantly increased c-Fos induction in various hypothalamic regions, namely, the medial preoptic area (MPO), periventricular nucleus (Pe), suprachiasmatic nucleus (SCN), supraoptic nucleus (SON), parvocellular division of the paraventricular nucleus (pPVN), anterior hypothalamic area (AH), arcuate nucleus (ARC) and posterior hypothalamic nucleus (PH). However, sub-LT caused no effect on c-Fos accumulation. This indicates that the hypothalamus responds uniquely to running in a threshold-like pattern distinct from the speed-dependent pattern previously reported for the medulla oblongata [Ohiwa et al., 2006a,b]. In addition, these results showed a physiologic basis for mild exercise useful for establishing our minimum running stress (MRS) rat model, or the running conditions that minimize the activation of the HPA axis.

Lack of association between genotype score and sprint/power performance in the Japanese population.

OBJECTIVES: This study aimed to examine the association between a total genotype score (TGS) based on previously published genetic polymorphism candidates and differences in sprint/power performance. DESIGN: Case-control association study. METHODS: We analysed 21 polymorphisms, which have previously been associated with sprint/power performance and related phenotypes, in 211 Japanese sprint/power track and field athletes (77 regional, 72 national, and 62 international athletes) and 649 Japanese controls using the TaqMan SNP genotyping assay. We calculated the TGS (maximum value of 100 for the theoretically optimal polygenic score) for the 21 polymorphisms. RESULTS: All groups exhibited similar TGSs (control: 55.9±7.2, regional: 55.1±7.1, national: 56.1±7.4, and international: 56.0±7.8, p=0.827 by one-way analysis of variance). Nine of the 21 polymorphisms had the same direction of effect (odds ratio >1.0) as in previous studies, while 12 had the opposite direction of effect (odds ratio <1.0). Three polymorphisms (rs699 in AGT, rs41274853 in CNTFR, and rs7832552 in TRHR), which had the same direction of effect as in previous studies, were associated with international sprint/power athlete status (p<0.05). However, after multiple testing corrections, the statistical significance of these polymorphisms was not retained. CONCLUSIONS: These results suggest that TGSs based on the 21 previously published sprint/power performance-associated polymorphisms did not influence the sprint/power athlete status of Japanese track and field athletes. However, our results maintain the possibility that three of these polymorphisms might be associated with sprint/power performance.

Differential responsiveness of c-Fos expression in the rat medulla oblongata to different treadmill running speeds.

Expression of the inducible transcription factor c-Fos was mapped in the rat medulla oblongata to identify the brain areas respond to different running speeds. Rats were subjected to 30 min of running, either at high speed, low speed or just sitting on a treadmill (control). Blood lactate levels were measured to confirm the physiological impact of different exercise intensities. The number of c-Fos-ir cells was counted and their spatial distributions were mapped through the rostral to the caudal level in the medulla. A statistically significant exercise intensity-dependent induction of c-Fos was observed in the nucleus of the solitary tract (NTS) and caudal ventrolateral medulla (CVL) in the medulla. Further, c-Fos induction was more predominant in the caudal part of each nucleus. The present data clearly show that different running speeds cause differential activation of each nucleus in the medulla, and in particular, the caudal parts of the NTS and the CVL are the most responsive to speed changes. The present study identifies brain areas newly found to be responsive to changes in running speed. These findings are likely to be particularly helpful in studies of specific neural circuits and their functions in response to different running speeds.

Activation of A1 and A2 noradrenergic neurons in response to running in the rat.

Since running accompanied with blood lactate accumulation stimulates the release of adrenocorticotropic hormone (ACTH), running above the lactate threshold (LT) acts as stress (running stress). To examine whether A1/A2 noradrenergic neurons that project to the hypothalamus activate under running stress, c-Fos immunohistochemistry was used to compare the effects of running with or without stress response on A1/A2 noradrenergic neurons. Blood lactate and plasma ACTH concentrations significantly increased in the running stress group, but not in the running without stress response and control groups, confirming different physiological impacts between different intensity of running with or without stress. Running stress markedly increased c-Fos accumulation in the A1/A2 noradrenergic neurons. Running without stress response also induced a significant increase in c-Fos expression in the A1/A2 noradrenergic neurons, and the percentage of the increase was smaller than that of running stress. The extent of c-Fos expression in the A1/A2 noradrenergic neurons correlates with exercise intensity, signifying that this neuronal activation is running speed-dependent. We thus suggest that A1/A2 noradrenergic neurons are activated in response to not only running stress, but also to other physiological running, enhanced by non-stressful running. These findings will be helpful in studies of specific neurocircuits and in identifying their functions in response to running at different intensities.

Increased c-fos gene expression in alpha motoneurons in rat loaded hindlimb muscles with inclined locomotion.

The potential usefulness of c-fos gene expression as an indicator of the activity level of spinal alpha motoneurons was examined in loaded locomotive rats. The motor pools of the plantaris (PL) and soleus muscles (SOL), mainly composed respectively of fast- and slow-twitch muscle fibers, were investigated in rats under locomotion at 25 m/min on a 20% incline. We first labeled motoneurons with a retrograde tracer, Nuclear Yellow (NY), and then quantified the c-fos mRNA expression level in the NY-labeled alpha motoneurons by means of in situ hybridization. Electromyographic (EMG) activities were also recorded. The c-fos expression level per alpha motoneuron showed a greater increase in the PL (75%) than in the SOL motor pool (38%). EMG activities also showed a greater increase in the PL (159%) than in the SOL (43%). Taken together, these results suggest that c-fos expression levels in alpha motoneurons are associated with the activity levels of their corresponding muscle. This cytochemical method for identifying the c-fos expression level has potential for use as a tool for estimating the activity level of large populations of alpha motoneurons in unrestricted animals.

Effects of systemic hypoxia on human muscular adaptations to resistance exercise training.

Hypoxia is an important modulator of endurance exercise-induced oxidative adaptations in skeletal muscle. However, whether hypoxia affects resistance exercise-induced muscle adaptations remains unknown. Here, we determined the effect of resistance exercise training under systemic hypoxia on muscular adaptations known to occur following both resistance and endurance exercise training, including muscle cross-sectional area (CSA), one-repetition maximum (1RM), muscular endurance, and makers of mitochondrial biogenesis and angiogenesis, such as peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), citrate synthase (CS) activity, nitric oxide synthase (NOS), vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1 (HIF-1), and capillary-to-fiber ratio. Sixteen healthy male subjects were randomly assigned to either a normoxic resistance training group (NRT, n = 7) or a hypoxic (14.4% oxygen) resistance training group (HRT, n = 9) and performed 8 weeks of resistance training. Blood and muscle biopsy samples were obtained before and after training. After training muscle CSA of the femoral region, 1RM for bench-press and leg-press, muscular endurance, and skeletal muscle VEGF protein levels significantly increased in both groups. The increase in muscular endurance was significantly higher in the HRT group. Plasma VEGF concentration and skeletal muscle capillary-to-fiber ratio were significantly higher in the HRT group than the NRT group following training. Our results suggest that, in addition to increases in muscle size and strength, HRT may also lead to increased muscular endurance and the promotion of angiogenesis in skeletal muscle.

Increased hemoglobin mass and VO2max with 10 h nightly simulated altitude at 3000 m.

PURPOSE: To quantify the changes of hemoglobin mass (Hbmass) and maximum oxygen consumption (VO2max) after 22 days training at 1300-1800 m combined with nightly exposure to 3000-m simulated altitude. We hypothesized that with simulated 3000-m altitude, an adequate beneficial dose could be as little as 10 h/24 h. METHODS: Fourteen male collegiate runners were equally divided into 2 groups: altitude (ALT) and control (CON). Both groups spent 22 days at 1300-1800 m. ALT spent 10 h/night for 21 nights in simulated altitude (3000 m), and CON stayed at 1300 m. VO2max and Hbmass were measured twice before and once after the intervention. Blood was collected for assessment of percent reticulocytes (%retics), serum erythropoietin (EPO), ferritin, and soluble transferrin receptor (sTfR) concentrations. RESULTS: Compared with CON there was an almost certain increase in absolute VO2max (8.6%, 90% confidence interval 4.8-12.6%) and a likely increase in absolute Hbmass (3.5%; 0.9-6.2%) at postintervention. The %retics were at least very likely higher in ALT than in CON throughout the 21 nights, and sTfR was also very likely higher in the ALT group until day 17. EPO of ALT was likely higher than that of CON on days 1 and 5 at altitude, whereas serum ferritin was likely lower in ALT than CON for most of the intervention. CONCLUSIONS: Together the combination of the natural and simulated altitude was a sufficient total dose of hypoxia to increase both Hbmass and VO2max.

Possible inhibitory role of prolactin-releasing peptide for ACTH release associated with running stress.

Exercise around the lactate threshold induces a stress response, defined as "running stress." We have previously demonstrated that running stress is associated with activation of certain regions of the brain, e.g., the paraventricular hypothalamic nucleus (PVN) and supraoptic nucleus, that are hypothesized to play an integral role in regulating stress-related responses, including ACTH release during running. Thus we investigated the role of prolactin-releasing peptide (PrRP), found in the ventrolateral medulla and the nucleus of the solitary tract, which is known to project to the PVN during running-induced ACTH release. Accumulation of c-Fos in PrRP neurons correlated with running speeds, reaching maximal levels under running stress. Intracerebroventricular injection of neutralizing anti-PrRP antibodies led to increased plasma ACTH level and blood lactate accumulation during running stress, but not during restraint stress. Exogenous intracerebroventricular administration of low doses of PrRP had the opposite effects. Therefore, our results suggest that, during running stress, PrRP-containing neurons are activated in an exercise intensity-dependent manner, and likewise the produced endogenous PrRP attenuates ACTH release and blood lactate accumulation during running stress. Here we provide a novel perspective on understanding of PrRP in the endocrine-metabolic response associated with running stress.