PhD (Biology), Senior Researcher of the Laboratory of Auxology E.A. Bondareva¹
Dr.Biol., Professor, Head of the Department of Anantomy and Biological Anthropology E.Z. Godina^2
1Institute and Museum of Anthropology, Lomonosov Moscow State University, Moscow
²Russian State University of Physical Education, Sport, Youth and Tourism, Moscow

Keywords: AMPD1, sambo wrestling, sports selection, sport genetics.

Introduction. The physical qualities of athletes depend to a considerable degree on individual genetic characteristics [13; 17]. One of the markers associated with physical efficiency is C34T polymorphism of adenosine monophosphate deaminase 1 (myoadenylate deaminase) or AMPD 1 gene [6; 8; 11]. It is activated after brief and intense muscular efforts and shifts the balance of reaction 2 ADP ↔ ATP + AMP towards the formation of ATP [14], activating reaction of AMP deamination to IMP in skeletal muscles [15]. This means that it affects indirectly the resynthesis of ATP at the time of submaximum and maximum intensity of physical efforts. In addition, the reaction of deamination is also the first reaction during the purine nucleotides cycle, which leads to utilization of adenine nucleotides and formation of the ammonium ion. The accumulation of IMP and ammonium stimulates the glycolytic activity of the skeletal muscles, which also leads to increased ATM concentration and helps skeletal muscles maintain high intensity of contractions [14; 15; 18]. The gene of AMP deaminase 1 is localized in the short arm of chromosome 1 (1р13). AMPD1 is expressed mainly in type II muscle fibers which ensure brief and intense muscle contractions. It is known that the substitution of cytosin for thymine in the second exon of AMPD1 (С34Т, rs17602729) leads to a substitution of glutamine codon for stop-codon, which causes premature termination of translation. Carriers of two mutant alleles AMPD1 (AMPD1*ТТ) practically lack this enzyme [18]. T allele is associated with muscle convulsions, muscle pains and early onset of fatigue during physical exercise [3; 18] and can be considered as a factor decreasing human physical capacities. However, T allele carriers suffering from cardiovascular diseases show the best survival rate and delayed development of disease as compared to CC genotype carriers [1].

Various groups of athletes were observed to assess the influence of this marker on success and selection in sports. Several studies have shown a decrease in the frequency of T allele in athletes from different kinds of sports [8; 16; 18]. In a group of football players, a statistically significant increase of the number of heterozygous carriers was revealed as compared to long-distance runners [9]. Also, heterozygous carriers show higher values of VO₂ max after creatine intake [10]. In a group of Bulgarian mountain climbers, who climbed Everest, the frequency of T allele is 30 %, exceeding a similar indicator in the general population and in other groups of athletes. Combined with increase in the frequencies of genotypes of gene II ACE and gene XX ACTN3 found in well trained mountain climbers, it is safe to assume that T allele AMPD1 is associated with an increase of endurance in athletes [4]. Summarizing the evidence presented in the literature, it can be concluded that C allele of gene AMPD1 allows the athletes to reach top performance in those sports where development of strength and strength and speed qualities is required. But the presence of mutant T allele AMPD1 is a factor, which determines the best adaptive resources of the cardiovascular system, at the same time decreasing the strength and speed capacities of the skeletal muscles.

Materials and Methods. 318 ethnic Russian males between 18 and 30 years of age participated in the research. 92 of them represented the reference group which was not involved in sport and 226 were sambo wrestlers. Samples of buccal epithelium were taken from athletes at the Russian Sambo championship of 2008 (Moscow). The participants in the reference group were examined at the facilities of Russian State University of Physical Education, Sport, Youth and Tourism (SCOLIPE). A sample of buccal epithelium was used as a biological material for genomic DNA isolation. The sampling of biological material was made using sterile urogenital probes (Type A "Universal"), China. After the collection of samples of biological material, the probes were dried and placed in a container, located in the hollow shaft of the probe, for transportation to the laboratory. The genotype of every participant in the study was determined on the basis of the polymorphous system of gene AMPD1 (С34Т, rs17602729). The genotyping was performed at the facilities of Litech LLC, Moscow. All tested subjects were informed about the purposes of the research and gave their informed consent. The statistical analysis of the collected data was performed using the software package Statistica 8.0. Non-parametric statistical test χ2 was used to evaluate the significance of the differences in the distribution of genotypes.

The study group included subjects with different ranks in sports at the moment of research: athletes of mass ranks, N=26 - CMS (Candidates for Masters of Sport), highly skilled athletes, N=141 - MS (Masters of Sport) and athletes of international class (N=59) - MSIC (Masters of Sport, International Class) and MMS (Merited Masters of Sport) (Table 1)

Results and Discussion. The genetic characteristics of an athlete are one of the factors which determines the success in a chosen sport [6; 13]. The existence of certain molecular genetic markers in the genome may not only positively influence the success in sports, but also limit it [5]. The С34Т substitution of the gene of AMP deaminase 1 is one of such markers. A lot of researches show a decrease in the frequency of mutant T allele (and TT genotype) in the athletes group compared to that of non-athletes [2; 8; 16; 19]. Table 1 shows the distribution of genotypes frequencies of the AMPD1 gene and in sambo wrestlers in total and in the reference group. The distribution of the frequencies in the reference group corresponds to the Hardy–Weinberg equilibrium (χ2 HW=1.75 p=0.18).

Table 1. Frequency distribution of genotypes of the AMPD1 gene in sambo wrestlers and in the reference group.

Some decrease (1.3 % versus 2.2 %) in the frequency of the homozygous TT genotype was observed in a studied sample of sambo wrestlers, but the analysis of the frequencies of genotypes between sambo wrestlers and the reference group did not detect statistically significant differences (χ2=1.36 p=0.50). Taking into consideration a certain decrease in the frequency of the TT genotype in the examined sample of athletes, a conclusion can be made about the presence of a strength and speed direction of the sample as a whole in the examined group of sambo wrestlers. The collected data correspond to the results of a research with Polish athletes active in sports which require first of all development of strength and speed physical performance [8]. The authors report a statistically significant decrease of the frequency of the TT genotype and T allele in this group of athletes, which indicates an advantage of the carrier of primary C allele which determines the normal activity level of the myoadenilatdeaminase [2; 6].

The distribution of genotypes in the reference group (Table 1) corresponds to the results received from other European samples. The examined sample of athletes is heterogeneous: it contains wrestlers who have different titles in sports which reflect the current level of sport qualification (Table 2). In this respect, it is safe to assume that the revealed trend of selection of carriers of primary AMPD1 C allele will strengthen with the increase of the level of sport qualification – from athletes of mass ranks (Candidates for Masters of Sports) to highly skilled athletes (Masters of Sports of International Class and Merited Masters of Sports). The entire sample of sambo wrestlers was divided into three subgroups according to their level of sport qualification (Table 2).

Table 2. Frequency distribution of genotypes of AMPD1 gene in subgroups of the examined sample according to the level of sports qualification of the tested subjects.

Statistically significant differences in the frequencies of genotypes in three formed subgroups as compared to the reference group were not found (χ2 =5.07 p=0.53). The collected data allows to suggest that the С34Т substitution of examined gene did not have a significant effect on the increase of sport skills in sambo despite the fact that the frequency of the genotype AMPD1*TT was lower in the group of highly skilled athletes than in the reference group (1.4% and 1.7% versus 2.2 %, accordingly). Similar results were obtained in the study of the distribution of AMPD1 genotypes in groups of highly skilled rowers and highly skilled athletes engaged in cyclic sports as compared to the non-athletes: no statistically significant differences were observed [12]. The research on the associations of С34Т substitution in the group of boat racers having different level of sport skills also did not reveal any statistically significant associations of different AMPD1 genotypes with the skill level in the tested subjects [7; 17].

Conclusion. As a whole, the examined group of sambo wrestlers showed some decrease in the occurrence of T allele and TT genotype, thus indicating some advantage of carriers of primary C allele associated with normal level of activity of the AMPD1 enzyme. At the same time, no statistically significant differences in the frequencies of AMPD1 genotypes and alleles with the increase of sport qualification (KMS, MS, MSMC, and MMS) were discovered. The obtained results permit to assume that the С34Т substitution of AMPD1 did not have any significant impact on the success in sambo wrestling.

Acknowledgements. The study was supported by the Russian Foundation for Basic Research (grant no. 16-06-00480).

References

1. Anderson J.L., Habashi J., Carlquist J.F., Muhlestein J.B., Horne B.D., Bair T.L., Pearson R.R., Hart N. A common variant of the AMPD1 gene predicts improved cardiovascular survival in patients with coronary artery disease // J. Am. Coll. Cardiol. – 2000. – Vol. 36, no. 4. –P. 1248 – 1252.
2. Cieszczyk P., Ostanek M., Leoсska-Duniec A., Sawczuk M., Maciejewska A., Eider J., Ficek K., Sygit K., Kotarska K. Distribution of the AMPD1 C34T polymorphism in Polish power-oriented athletes // J. Sports Sci. – 2012. – Vol. 30, no. 1. – P. 31 – 35.
3. Colombini A., Lombardi G., Banfi G., Arpesella M., Pelissero G. Athleticogenomics and elite athletes: a review of the state of the art and a possible relationship with inflammatory response // IJPH. – 2011. – Vol. 8, no. 3. – P. 275 – 285.
4. Djarova T., Bardarev D., Boyanov D., Kaneva R., Atanasov P. Performance enhancing genetic variants, oxygen uptake, heart rate, blood pressure and body mass index of elite high altitude mountaineers // Acta Physiol. Hung. – 2013. – Vol. 100, no. 3. – P. 289 – 301.
5. Eynon N., Hanson E.D., Lucia A., Houweling P.J., Garton F., North K.N., Bishop D.J. Genes for elite power and sprint performance: ACTN3 leads the way // Sports Med. – 2013. – Vol. 43, no. 9. – P. 803 – 817.
6. Fedotovskaya O.N., Danilova A.A., Akhmetov I.I. Effect of AMPD1 gene polymorphism on muscle activity in humans // Bull. Exp. Biol. Med. – 2013. – Vol. 154, no. 4. – P. 489 – 491.
7. Fischer H., Esbjцrnsson M., Sabina R.L., Strцmberg A., Peyrard-Janvid M., Norman B. AMP deaminase deficiency is associated with lower sprint cycling performance in healthy subjects // J. Appl. Physiol. – 2007. Vol. 103. P. 315 – 322.
8. Ginevičienė V., Jakaitienė A., Pranculis A., Milašius K., Tubelis L. and Utkus A. AMPD1 rs17602729 is associated with physical performance of sprint and power in elite Lithuanian athletes // BMC Genetics. 2014. – Vol. 15. – P. 58 – 62.
9. Juffer P., Furrer R., Gonzбlez-Freire M., Santiago C., Verde Z., Serratosa L., Morate F.J., Rubio J.C., Martin M.A., Ruiz J.R., Arenas J., Gуmez-Gallego F., Lucia A. Genotype distributions in top-level soccer players: a role for ACE? // Int. J. Sports Med. – 2009. – Vol. 30, no. 5. – P. 387 – 392.
10. Lifanov D., Khadyeva M.N., Rahmatullina L.Sh., Demenev S.V., Ibragimov R.R. Effect of creatine supplementation on physical performance are related to the AMPD1 and PPARG genes polymorphisms in football players // Ross. Fiziol. Zh. Im I.M. Sechenova. – 2014. – Vol. 100, no. 6. – P. 767 – 776.
11. Lippi G., Longo U.G., Maffulli N. Genetics and sports // Br. Med. Bull. – 2009. – Vol. 7, no. 1. P. 2 – 21.
12. Muniesa C.A., Gonzбlez-Freire M., Santiago C., Lao J.I., Buxens A., Rubio J.C., Martнn M.A., Arenas J., Gomez-Gallego F., Lucia A. World-class performance in lightweight rowing: is it genetically influenced? A comparison with cyclists, runners and non-athletes // Br. J. Sports Med. – 2010. – Vol. 44, no. 12. – P. 898 – 901.
13. Myburgh K.H. What makes an endurance athlete world-class? Not simply a physiological conundrum // Comp. Biochem. Physiol. A Mol. Integr. Physiol. – 2003. – Vol. 136, no. 5. – p. 171–190.
14. Norman B., Mahnke-Zizelman D.K., Vallis A., Sabina R.L. Genetic and other determinants of AMP deaminase activity in healthy adult skeletal muscle // J. Appl. Physiol. – 1998. – Vol. 85. – P. 1273 – 1278.
15. Rico-Sanz J., Rankinen T., Joanisse D.R., Leon A.S., Skinner J.S., Wimore J.H., Rao D.C., Bouchard C. Associations between cardiorespiratory responses to exercise and the C34T AMPD1 gene polymorphism in the HERITAGE Family Study // Physiol. Genomics. – 2003. –no. 14. – P. 161 – 166.
16. Rubio J.C., Martin M.A., Rabadan M., Gomez-Gallego F., San Juan A.F., Alonso J.M., Chicharro J.L., Perez M., Arenas J., Lucia A. Frequency of the C34T mutation of the AMPD1 gene in world-class endurance athletes: does this mutation impair performance? // J. Appl. Physiol. – 2005. – no. 98. – P. 2108 – 2112.
17. Santiago C., Ruiz J.R., Muniesa C.A., Gonzбlez-Freire M., Gуmez-Gallego F., Lucia A. Does the polygenic profile determine the potential for becoming a world-class athlete? Insights from the sport of rowing // Scand. J. Med. Sci. Sports. – 2010. – Vol. 20, no. 1. – e188-194.
18. Tarnopolsky M.A., Parise G., Gibala M.J., Graham T., Rush J.W. Myoadenylatedeaminase deficiency does not affect muscle anaplerosis during exhaustive exercise in humans // J. Physiol. – 2001. – Vol. 533. – P. 881–889.
19. Tsianos G.I., Evangelou E., Boot A., M. Zillikens M.C., van Meurs J.B., Uitterlinden A.G., and Ioannidis J.P. Associations of polymorphisms of eight muscle- or metabolism-related genes with performance in Mount Olympus marathon runners // J. Appl. Physiol. – 2010. – Vol. 108. – P. 567–574.

Corresponding author: Bondareva.E@gmail.com

Abstract. The study is devoted to C34T polymorphism of adenosine monophosphate deaminase 1 (myoadenylate deaminase) or AMPD 1 gene associated with physical efficiency in humans. Various groups of athletes were observed to assess the influence of this marker on success and selection in sports. 318 ethnic Russian males between 18 and 30 years of age participated in the research. 92 of them represented the reference group which was not involved in sport and 226 sambo wrestlers. Samples of buccal epithelium were taken from athletes at the Russian Sambo championship of 2008 (Moscow). As a whole, the examined group of sambo wrestlers showed some decrease in the occurrence of T allele and TT genotype, thus indicating some advantage of carriers of primary C allele associated with the normal level of activity of the AMPD1 enzyme. At the same time, no statistically significant differences in the frequencies of AMPD1 genotypes and alleles with the increase of sports qualification (KMS, MS, MSMC, and MMS) were discovered.