Journal of Strength & Conditioning Research:
The ACTN3 R577X Polymorphism Is Associated With Muscle Power in Male Japanese Athletes
Kikuchi, Naoki1; Nakazato, Koichi2; Min, Seok-ki2; Ueda, Dai3; Igawa, Shoji4
1Sports Training Center, Nippon Sport Science University, Tokyo, Japan;
2Laboratory of Exercise Physiology, Nippon Sport Science University, Tokyo, Japan;
3Faculty of Health and Nutrition, Bunkyo University, Kanagawa, Japan; and
4Laboratory of Sports Nutrition, Nippon Sport Science University, Tokyo, Japan
Address correspondence to Naoki Kikuchi, email@example.com.
Abstract: Kikuchi, N, Nakazato, K, Min, S-k, Ueda, D, and Igawa, S. The ACTN3 R577X polymorphism is associated with muscle power in male Japanese athletes. J Strength Cond Res 28(7): 1783–1789, 2014—In this study, we investigated whether the ACTN3 R577X polymorphism is associated with muscular power in Japanese collegiate athletes by analyzing the mean and peak power results of a 30-second Wingate anaerobic test (WAnT) with respect to the ACTN3 R577X genotype in 253 Japanese athletes (144 men and 109 women). Each athlete performed a 30-second WAnT with a resistance equal to 7.5% of his or her body weight. Genotyping for the ACTN3 R577X (rs1815739) polymorphism was performed using the TaqMan approach. The ACTN3 R577X genotypes exhibited a Hardy-Weinberg equilibrium distribution in our population. The relative and absolute mean power results of the 30-second WAnT did not differ significantly among the genotypes. However, the relative peak power result of the WAnT was significantly higher in the R-allele-dominant model groups than in the XX group in male but not female athletes. These results suggest that the ACTN3 R allele is associated with the relative peak power during the WAnT in male Japanese collegiate athletes.
Elite athletic performance is a complex phenotype determined by several environmental factors, including dietary, physical training, and social influences. Genetic variation may also contribute to interindividual differences in athletic performance, and a recent review noted that more than variants of 200 genes are associated with fitness-related phenotypes (20). One of the most potent of these, the α-actinin-3 (ACTN3) R577X polymorphism, has been associated with muscle fiber composition (22), muscle strength (14), and elite performance (8,10,25).
The ACTN3 R allele and RR genotype have been found to be associated with top-level power-oriented athletic performance in a broad variety of ethnic groups, including Israelis (5), Finns (17), Greeks (18), Russians (4), and Japanese (12). In contrast, previous studies in whites found that the ACTN3 XX genotype confers better endurance performance in the general population (21,25). Although previous studies (1) reported that the ACTN3 RR genotype is associated with endurance performance, the role of the XX genotype in endurance performance remains unclear.
The relationship between the ACTN3 R577X genotype and muscular phenotype has been examined in the general population in several ethnic groups. Women with the XX genotype have lower baseline elbow flexor isometric strength (3), lower knee extensor peak torque (24), and a smaller thigh muscle (26) in comparison with women with RR genotype. Similar associations have been observed in competitive athletes. We recently confirmed a linear correlation between the frequency of the ACTN3 R allele (RR or RX) and actual competition results in Japanese wrestlers (12), as reported by others (4). Taken together, there appears to be a relationship between the ACTN3 genotype and muscle function.
Previous studies (7,15) did not specifically examine the relationship between ACTN3 genotype and anaerobic performance, especially in trained athletes. Anaerobic performance is generally estimated by tests such as the vertical jump and the Wingate test, and such information helps athletes and their coaches evaluate and adjust their conditioning programs. The Wingate anaerobic power test (WAnT) in particular is frequently used to measure anaerobic performance (13,27). The peak power (the highest-power performance during any 5-second period) and mean power (the total power performance during the entire 30-second period) are traditionally considered good general indices of anaerobic power and capacity. The WAnT has been confirmed to be an accurate indicator of athletic performance (13). Exploration of the association between the ACTN3 genotype and the actual anaerobic performance phenotype is important to help trained athletes, especially those in power-oriented sports, to design more suitable individualized training programs. Despite this importance, the WAnT was not employed in the previous studies (7,15) concerning the specific role of the ACTN3 genotype in trained athletes.
The purpose of this study was to examine the relationship between the ACTN3 R577X polymorphism genotype and the athletic phenotype in Japanese collegiate athletes, using the WAnT to estimate each participant's maximum anaerobic capacity.
Experimental Approach to the Problem
We used a cross-sectional design to examine the association between the ACTN3 R577X polymorphism and WAnT results (mean power and peak power) in 253 Japanese trained athletes. The experimental period was from July through August 2012. All subjects were in the preseason training phase for their respective sports. Each subject performed a 30-second WAnT using a resistance equal to 7.5% of his or her body weight. All of the subjects were involved in top-level intercollegiate sports and had extensive weight training experience. Each subject also completed a questionnaire concerning his or her training status and athletic experience.
We enrolled 253 Japanese collegiate athletes (144 men and 109 women; age range, 18–22 years) in this study. The subjects' general characteristics and specific sports events are shown in Table 1. The participants were informed of the purpose and method of the study to ensure that they understood completely, and each provided written informed consent to participate. The study was approved by the ethics committee of Nippon Sport Science University in Japan (010-G01) and was conducted in accordance with the Declaration of Helsinki for Human Research.
Genotyping of a DNA polymorphism in the ACTN3 gene was performed in 253 Japanese collegiate athletes. DNA samples were obtained from the subjects' buccal cells by rubbing the inner surface of each subject's mouth with a cotton swab. After collection, the cells were lysed in 50 µl of lysis solution (20 mmol·L−1 Tris-HCI (pH 8.0) containing 5 mmol·L−1 ethylenediaminetetraacetic acid, 400 mmol·L−1 NaCI, 0.3% sodium dodecyl sulfate, and 10 mg·ml−1 proteinase K) and incubated at 55° C for 30 minutes. The samples were then stored at 4° C until polymerase chain reaction (PCR). Genotyping for the ACTN3 R577X polymorphism was performed by real-time PCR using a TaqMan probe (rs1815739, Pre-Designed SNP Genotyping assays; Applied Biosystems, Foster City, CA, USA). Polymerase chain reaction cycling was performed using a heat block (CFD-3120J1, BioRad, Hercules, CA, USA) as follows: an initial melting step at 95° C for 10 minutes followed by 39 cycles consisting of 92° C for 15 seconds and 60° C for 1 minute and then a melting curve from 65° C to 95° C (3).
The WAnT was performed on a PowerMaxV II (Combi, Tokyo, Japan) using a resistance equal to 7.5% of the athlete's body weight for both the male and female athletes. Each subject first performed a 3–5 minute warm-up on a cycle ergometer in which he or she strived to achieve a warm-up heart rate of 130–140 b·min−1. We evaluated the mean power and peak power during the 30-second test. All subjects performed the WAnT and underwent DNA sampling at the same time of the experimental day. The experimental period was from July through August 2012.
The SPSS statistical package version 16.0 for Windows (SPSS, Inc., Chicago, IL, USA) was used to perform all statistical evaluations. Allele frequencies were determined by gene counting. Pearson's χ2 test and Fisher's exact test were used to confirm that the observed genotype frequencies exhibited a Hardy-Weinberg equilibrium distribution. Differences in the mean and peak power results of the WAnT among the ACTN3 R577X genotypes were tested using analysis of variance (with Tukey's multiple comparison test) and between the XX and R-dominant model (RR and RX genotypes) groups using unpaired t-tests. Linear regression analysis was performed to estimate the degree of variance in the mean and peak power results of the WAnT associated with the ACTN3 R577X genotype. The enough number of subjects was determined by a sample size estimation using the data from previous studies that examine the relationship between ACTN3 genotype and muscle phenotype (13,27). The estimation was based on the effect size of 0.3, alpha level of 0.05, and a power (1-β) of 0.80. Statistical calculation was performed by G*power (6). We confirmed that the sample size was enough for the design of this study. The level of significance was set at p ≤ 0.05.
The ACTN3 genotypes exhibited a Hardy-Weinberg equilibrium distribution among athletes (all athletes: χ2 = 0.26 and p = 0.702; male athletes: χ2 = 0.004 and p = 0.944; female athletes: χ2 = 1.44 and p = 0.312). The genotype frequencies and the allele frequency of the ACTN3 R577X polymorphism are shown in Table 2. None of the subjects' characteristics differed significantly among the ACTN3 R577X genotypes.
The anaerobic power (absolute/relative) results of the WAnT with respect to the ACTN3 R577X genotype and the R-dominant model in male athletes are shown in Table 3. The relative WAnT peak power differed significantly among the ACTN3 genotypes: athletes with the RR genotype had a significantly higher relative peak power than those with the RX (p = 0.039) or XX (p = 0.024) genotype. The R-dominant model (RR and RX genotypes) showed significantly higher relative peak power than the XX genotype (p = 0.045). Among the female athletes, however, the WAnT power did not differ significantly among the ACTN3 R577X genotypes (Table 4). The ACTN3 R577X genotype accounted for 4.6% of the variability in the relative peak power result of the WAnT among the male athletes (p = 0.006).
To the best of our knowledge, this study included the largest number of Japanese athletes among similar studies performed to date and therefore provides useful insight into the role of the ACTN3 R577X genotype in trained individuals. Our major finding is that the ACTN3 R allele, especially when homozygous, influences the relative WAnT peak power of Japanese male athletes. We also showed by regression analysis that the ACTN3 R577X genotype accounts for approximately 4.6% of the variance in the relative peak power result of the WAnT in Japanese male athletes.
Two major energy sources are required during the WAnT. The first is the adenosine triphosphate-phosphocreatine system, which lasts only for 3–15 seconds during maximum effort. The second system is anaerobic glycolysis, which can be sustained for the remainder of the all-out effort. The peak power recorded is the maximal power output achieved for 5 seconds, usually the first 5 seconds, of the WAnT (27). We found that the ACTN3 R allele was associated with the relative peak power result of the WAnT in the male Japanese athletes. Because the peak power was observed within the first 5 seconds of the test in all subjects, this result suggests that athletes with the R allele may possess a higher-capacity phosphagen system, and we cannot exclude the possibility that the glycolytic system is also affected by the ACTN3 genotype.
We also found that 4.6% of the variability in the relative peak power result of the WAnT among the male Japanese athletes was attributable to the ACTN3 R577X genotype. A previous study on the general population reported that the ACTN3 R577X polymorphism is responsible for 1–2% of the variation in muscle strength (3,24). However, no association between the ACTN3 R577X genotype and the muscle power results of the WAnT was reported in individuals who were active but not athletes (9,22). On the other hand, Massidda et al. (15) reported that the ACTN3 R577X genotype accounted for 8.0% of the variation in squat-jump performance in elite soccer players. These results suggest that the contribution of the ACTN3 R577X genotype becomes more apparent in competitive athletes, as we had hypothesized. We propose that the contribution of the ACTN3 R allele manifests during strenuous sporting activities, as shown in this study.
This study detected a significant relationship between the ACTN3 R577X genotype and muscle power only in male athletes. Because the relationship between the ACTN3 R577X genotype and the muscle phenotype with respect to sex has not previously been examined in the Japanese population, we believe that our results are the first documented evidence for a sex-dependent effect of the ACTN3 R/X polymorphism. Previous studies did show that the absence of α-actinin-3 protein negatively influences the peak isokinetic torque during knee extension in middle-aged women but not men (24). Moran et al. (16) reported that men with the R allele (RR or RX) had better sprinting ability than those with the XX genotype. These results support the possibility of sex-dependent effects of the ACTN3 R577X genotype on muscle phenotype and athletic performance.
The possible mechanisms underlying the association between the ACTN3 R577X polymorphism and power-oriented performance have been discussed in detail elsewhere. Recent findings have indicated that the percentage of surface coverage and the number of type IIx fibers is greater in individuals with the RR genotype group than in those with the XX genotype among both athletes (2) and nonathletes (20).
In this study, we examined the relationship between a single genetic factor and anaerobic performance in Japanese athletes. We also recently showed a significant relationship between a combination of 2 polymorphisms (ACTN3 R577X and ACE I/D) and elite performance (11). However, a comprehensive understanding requires consideration of potential confounding factors such as other polymorphisms and environmental factors.
In summary, we found a positive relationship between the ACTN3 R577X genotype and the relative peak power result of the WAnT in male Japanese athletes. Our data indicate that the ACTN3 R allele (RR and RX genotypes) is associated with power output capacity.
The ACTN3 R577X genotype can provide useful information (e.g., talent selection and genotype-based customization for training) for athletes, especially well-trained men and their strength and conditioning coaches and sports coaches. According to our previous study (12), athletes with the ACTN3 R allele (RR and RX) have an advantage over XX individuals in terms of power-oriented performance. Furthermore, Vincent et al. (23) described a protective role of the α-actinin-3 protein (completely absent in XX individuals) against muscle damage after eccentric training. In addition, professional soccer players with the XX genotype exhibited higher CK activities, α-actin concentrations, and levels of cortisol than did their RR counterparts (19).
Therefore, individuals with the XX genotype, being completely deficient in the α-actinin-3 protein, exhibit inferior skeletal muscle function in terms of force generation from contraction or a poor ability to recover from high-intensity intermittent exercise. These factors might predict some aspects of response to training and thus provide useful information for strength and conditioning coaches.
1. Ahmetov II, Druzhevskaya AM, Astratenkova IV, Popov DV, Vinogradova OL, Rogozkin VA. The ACTN3 R577X polymorphism in Russian endurance athletes. Br J Sports Med 44: 649–652, 2010.
2. Ahmetov II, Druzhevskaya AM, Lyubaeva EV, Popov DV, Vinogradova OL, Williams AG. The dependence of preferred competitive racing distance on muscle fibre type composition and ACTN3 genotype in speed skaters. Exp Physiol 96: 1302–1310, 2011.
3. Clarkson PM, Devaney JM, Gordish-Dressman H, Thompson PD, Hubal MJ, Urso M, Price TB, Angelopoulos TJ, Gordon PM, Moyna NM, Pescatello LS, Visich PS, Zoeller RF, Seip RL, Hoffman EP. ACTN3 genotype is associated with increases in muscle strength in response to resistance training in women. J Appl Physiol (1985) 99: 154–163, 2005.
4. Druzhevskaya AM, Ahmetov II, Astratenkova IV, Rogozkin VA. Association of the ACTN3 R577X polymorphism with power athlete status in Russians. Eur J Appl Physiol 103: 631–634, 2008.
5. Eynon N, Duarte JA, Oliveira J, Sagiv M, Yamin C, Meckel Y, Goldhammer E. ACTN3 R577X polymorphism and Israeli top-level athletes. Int J Sports Med 30: 695–698, 2009.
6. Faul F, Erdfelder E, Lang AG, Buchner A. G*Power 3: A flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods 39: 175–191, 2007.
7. Garatachea N, Verde Z, Santos-Lozano A, Yvert T, Rodriguez-Romo G, Sarasa FJ, Hernandez-Sanchez S, Santiago C, Lucia A. ACTN3 R577X polymorphism and explosive leg muscle power in elite basketball players. Int J Sports Physiol Perform 2013.
8. Gomez-Gallego F, Santiago C, Gonzalez-Freire M, Muniesa CA, Fernandez Del Valle M, Perez M, Foster C, Lucia A. Endurance performance: Genes or gene combinations? Int J Sports Med 30: 66–72, 2009.
9. Hanson ED, Ludlow AT, Sheaff AK, Park J, Roth SM. ACTN3 genotype does not influence muscle power. Int J Sports Med 31: 834–838, 2010.
10. Juffer P, Furrer R, Gonzalez-Freire M, Santiago C, Verde Z, Serratosa L, Morate FJ, Rubio JC, Martin MA, Ruiz JR, Arenas J, Gomez-Gallego F, Lucia A. Genotype distributions in top-level soccer players: A role for ACE? Int J Sports Med 30: 387–392, 2009.
11. Kikuchi N, Min SK, Ueda D, Igawa S, Nakazato K. Higher frequency of the ACTN3 R allele + ACE DD genotype in Japanese elite wrestlers. J Strength Cond Res 26: 3275–3280, 2012.
12. Kikuchi N, Ueda D, Min SK, Nakazato K, Igawa S. The ACTN3 XX genotype's underrepresentation in Japanese elite wrestlers. Int J Sports Physiol Perform 8: 57–61, 2013.
13. Legaz-Arrese A, Munguia-Izquierdo D, Carranza-Garcia LE, Torres-Davila CG. Validity of the Wingate anaerobic test for the evaluation of elite runners. J Strength Cond Res 25: 819–824, 2011.
14. MacArthur DG, Seto JT, Chan S, Quinlan KG, Raftery JM, Turner N, Nicholson MD, Kee AJ, Hardeman EC, Gunning PW, Cooney GJ, Head SI, Yang N, North KN. An Actn3 knockout mouse provides mechanistic insights into the association between alpha-actinin-3 deficiency and human athletic performance. Hum Mol Genet 17: 1076–1086, 2008.
15. Massidda M, Corrias L, Ibba G, Scorcu M, Vona G, Calo CM. Genetic markers and explosive leg-muscle strength in elite Italian soccer players. J Sports Med Phys Fitness 52: 328–334, 2012.
16. Moran CN, Yang N, Bailey ME, Tsiokanos A, Jamurtas A, MacArthur DG, North K, Pitsiladis YP, Wilson RH. Association analysis of the ACTN3 R577X polymorphism and complex quantitative body composition and performance phenotypes in adolescent Greeks. Eur J Hum Genet 15: 88–93, 2007.
17. Niemi AK, Majamaa K. Mitochondrial DNA and ACTN3 genotypes in Finnish elite endurance and sprint athletes. Eur J Hum Genet 13: 965–969, 2005.
18. Papadimitriou ID, Papadopoulos C, Kouvatsi A, Triantaphyllidis C. The ACTN3 gene in elite Greek track and field athletes. Int J Sports Med 29: 352–355, 2008.
19. Pimenta EM, Coelho DB, Cruz IR, Morandi RF, Veneroso CE, de Azambuja Pussieldi G, Carvalho MR, Silami-Garcia E, De Paz Fernandez JA. The ACTN3 genotype in soccer players in response to acute eccentric training. Eur J Appl Physiol 112: 1495–1503, 2012.
20. Rankinen T, Roth SM, Bray MS, Loos R, Perusse L, Wolfarth B, Hagberg JM, Bouchard C. Advances in exercise, fitness, and performance genomics. Med Sci Sports Exerc 42: 835–846, 2010.
21. Shang X, Huang C, Chang Q, Zhang L, Huang T. Association between the ACTN3 R577X polymorphism and female endurance athletes in China. Int J Sports Med 31: 913–916, 2010.
22. Vincent B, De Bock K, Ramaekers M, Van den Eede E, Van Leemputte M, Hespel P, Thomis MA. ACTN3 (R577X) genotype is associated with fiber type distribution. Physiol Genomics 32: 58–63, 2007.
23. Vincent B, Windelinckx A, Nielens H, Ramaekers M, Van Leemputte M, Hespel P, Thomis MA. Protective role of alpha-actinin-3 in the response to an acute eccentric exercise bout. J Appl Physiol (1985) 109: 564–573, 2010.
24. Walsh S, Liu D, Metter EJ, Ferrucci L, Roth SM. ACTN3 genotype is associated with muscle phenotypes in women across the adult age span. J Appl Physiol (1985) 105: 1486–1491, 2008.
25. Yang N, MacArthur DG, Gulbin JP, Hahn AG, Beggs AH, Easteal S, North K. ACTN3 genotype is associated with human elite athletic performance. Am J Hum Genet 73: 627–631, 2003.
26. Zempo H, Tanabe K, Murakami H, Iemitsu M, Maeda S, Kuno S. ACTN3 polymorphism affects thigh muscle area. Int J Sports Med 31: 138–142, 2010.
27. Zupan MF, Arata AW, Dawson LH, Wile AL, Payn TL, Hannon ME. Wingate Anaerobic Test peak power and anaerobic capacity classifications for men and women intercollegiate athletes. J Strength Cond Res 23: 2598–2604, 2009.
gene polymorphism; anaerobic power; Wingate test; collegiate athlete
Copyright © 2014 by the National Strength & Conditioning Association.
What does "Remember me" mean?
By checking this box, you'll stay logged in until you logout. You'll get easier access to your articles, collections,
media, and all your other content, even if you close your browser or shut down your
To protect your most sensitive data and activities (like changing your password),
we'll ask you to re-enter your password when you access these services.
What if I'm on a computer that I share with others?
If you're using a public computer or you share this computer with others, we recommend
that you uncheck the "Remember me" box.
Highlight selected keywords in the article text.
Data is temporarily unavailable. Please try again soon.
Readers Of this Article Also Read