Dr.Med. Dr. Med. A.V. Kalinin^{1, 3}
O.A. Klever-Chekunova¹
M.K. Chekunov¹
Dr. Biol. E.M. Chekunova^2
1Lesgaft National State University of Physical Education, Sport and Health, St. Petersburg
²Saint Petersburg State University, Saint Petersburg
³Saint Petersburg State Pediatric Medical University, Saint Petersburg

Corresponding author: alatir.company@gmail.com

Abstract

Objective of the study was to conduct a comparative analysis of the frequency of polymorphic alleles (SNP) of the AMPD1 (rs17602729), COL1A1 (rs1800012), and COL5A1 (rs12722) genes in successful figure skaters and children, who had not previously trained or taken part in sports competitions and were still selected for sports sections.

Methods and structure of the study. The experiment was run using the materials collected during the implementation of the project of ANO "Stan Chempionom" at Lesgaft National State University of Physical Education, Sport and Health, St. Petersburg. 13 figure skaters of different levels of sports qualification were genotyped in terms of the above-mentioned three polymorphisms: 6 Masters of Sport, 2 Candidate Masters of Sport, 4 I Class athletes, and 1 III Class athlete. The Control Group (CG) was made of 130 children still being selected for sports sections, who had been subjected to the DNA tests under the project of ANO "Stan Chempionom".

It should be immediately noted that due to the small number of representatives of the elite sport, the authors did not differentiate between the figure skating disciplines, nor were the groups of participants differentiated by gender. The subjects’ age (group of athletes and CG subjects jointly) ranged from 6 to 24 years.

The biological material for DNA testing was collected by scraping the epithelial cells of the oral cavity using special sterile disposable probes. Each subject was asked to rub the buccal cavity with the tip of the probe for 10-15 seconds. DNA isolation from buccal epithelial cells was made using the modified STAB method.

Results and conclusions. The findings show that in the sport like figure skating, the following ancestral alleles predominate in the genotypes in athletes who regularly train and successfully increase their sports skill and qualification level: C of the AMPD1 gene (rs17602729), G of the COL1A1 gene (rs1800012), C of the COL5A1 gene (rs12722). This fact suggests that the results obtained in the group of top-class athletes are not random and are supported by the physiological expression of the nature of the above-mentioned genes.

Consequently, the findings indicate association of the AMPD1 (rs17602729), COL1A1 (rs1800012), and COL5A1 (rs12722) gene polymorphisms with successful trainings of athletes in figure skating sections. The assumption that the sample results are conditioned by the similarity in athletes’ physiology is interesting, and the authors plan to continue this study by increasing the sample of top-class athletes (HMS and WCMS).

Keywords: figure skating, sports selection, genes, polymorphisms (SNP), allele, PCR, DNA, sports, predisposition, AMPD1, COL1A1, COL5A1.

Background. Determining the performance of a future athlete at a young age is a difficult task, and the peak of loading falls on the top sports mastery stage, which coincides with the period of puberty (all hereditary physiological features of a young athlete come into sharp focus in adolescence) [1, 3, 5]. Among the many important physiological indicators of an athlete’s success, the authors chose such inherited traits as the ability to recover between trainings and the presence or absence of a predisposition to increased traumatism. In top-class sports, the following gene polymorphisms were studied to determine these traits: AMPD1 gene (rs17602729), COL1A1 gene (rs1800012), and COL5A1 gene (rs12722):

• The AMPD1 gene encodes adenosine monophosphate deaminase 1, a skeletal muscle protein, which is important for energy metabolism [4, 8, 9]. The rs17602729 polymorphism (C>T) is a replacement for cytosine (C) by thymine (T) at position 34 (C34T) of the AMPD1 gene. As a result of this substitution, the codon representing the amino acid glutamine is replaced by the stop codon, which results in the synthesis of a shortened protein without catalytic activity [9].
• The COL1A1 gene encodes the pro-alpha 1 chain of type I collagen, which is an important structural component of connective tissue [4]. The studied polymorphism rs1800012 is the replacement of guanine (G) by thymine (T) in the regulatory region of the COL1A1 gene at position 1245 (G1245T). As a result of this nucleotide replacement, the gene transcription character is altered and the normal relation of collagen chains in the structure of collagen fibers is disturbed. The presence of allele T in the genotype (rs1800012 of the COL1A1 gene) is associated with a decrease in the mineral density of the bones, higher chance of development of osteoporosis and bone fractures, as well as with ligament sprain due to active trainings [8].
• The COL5A1 gene encoding the alpha 1 chain of type V collagen is also an important structural component of connective tissue [4]. The rs12722 polymorphism is associated with a predisposition to the development of chronic Achilles tendinosis, as well as with the increased frequency of cruciate ligament ruptures [10]. This polymorphism is the result of substitution in the sequence of nucleotides: cytosine (C) by thymine (T), at position 267 of the non-coding region 3'-UTR of the COL5A1 gene (C267T), which results in changes in the mRNA stability of COL5A1 within normal physiological oscillations and does not disrupt the formation of collagen fibrils [11].

Objective of the study was to conduct a comparative analysis of the frequency of polymorphic alleles (SNP) of the AMPD1 (rs17602729), COL1A1 (rs1800012), and COL5A1 (rs12722) genes in successful figure skaters and children, who had not previously trained or taken part in sports competitions and were still selected for sports sections.

Methods and structure of the study. The experiment was run using the materials collected during the implementation of the project of ANO "Stan Chempionom" at Lesgaft National State University of Physical Education, Sport and Health, St. Petersburg. 13 figure skaters of different levels of sports qualification were genotyped in terms of the above-mentioned three polymorphisms: 6 Masters of Sport, 2 Candidate Masters of Sport, 4 I Class athletes, and 1 III Class athlete. The Control Group (CG) was made of 130 children still being selected for sports sections, who had been subjected to the DNA tests under the project of ANO "Stan Chempionom".

It should be immediately noted that due to the small number of representatives of the high-profile sport, the authors did not differentiate between the figure skating disciplines, nor were the groups of participants differentiated by gender. The subjects’ age (group of athletes and CG subjects jointly) ranged from 6 to 24 years.

The biological material for DNA testing was collected by scraping the epithelial cells of the oral cavity using special sterile disposable probes. Each subject was asked to rub the buccal cavity with the tip of the probe for 10-15 seconds. DNA isolation from buccal epithelial cells was made using the modified STAB method [6]. The AMPD1 (rs17602729), COL1A1 (rs1800012), and COL5A1 (rs12722) genes polymorphisms (SNP - Single Nucleotide Polymorphisms) were detected by the method of real-time allele-specific polymerase chain reactions (PCR-RT). All reactions were detected using the BioRad CFX-96 device; the primers were designed during work. Nucleotide sequences within amplified DNA fragments were tested by the direct Sanger sequencing on the automatic capillary sequencer ABI PRISM 3130XL. When analyzing the results obtained in the laboratory studies, the frequency of the alleles and genotypes under study was evaluated. The statistical significance of differences between the study groups was determined using Fisher’s Exact Test (p<0.05).

Results and discussion. The genotype test results demonstrated by the two groups showed no genotypes that were homozygous for mutant alleles in the group of figure skaters: TT of the AMPD1 gene (rs17602729), TT of the COL1A1 gene (rs1800012), TT of the COL5A1 gene (rs12722); heterozygous genotypes of these genes were under-represented, and for the COL1A1 gene (rs1800012) were absent at all. In CG, the proportion of such genotypes ranged from 20.7% to 28.5% (Table 1).

Table 1. Genotype test results of figure skaters and children selected for sports section

Table 2. Compositional data on distribution of genotypes and alleles in two study groups

• The values p<0.05, indicating statistically significant differences in the allele and genotype frequencies between the figure skaters and CG children, are highlighted in bold.

Conclusions. The findings show that in the sport like figure skating, the following ancestral alleles predominate in the genotypes in athletes who regularly train and successfully increase their sports skill and qualification level: C of the AMPD1 gene (rs17602729), G of the COL1A1 gene (rs1800012), C of the COL5A1 gene (rs12722). This fact suggests that the results obtained in the group of top-class athletes are not random and are supported by the physiological expression of the nature of the above-mentioned genes.

Consequently, the findings indicate association of the AMPD1 (rs17602729), COL1A1 (rs1800012), and COL5A1 (rs12722) genes polymorphisms with successful trainings of athletes in figure skating sections. The assumption that the sample results are conditioned by the similarity in athletes’ physiology is interesting, and the authors plan to continue this study by increasing the sample of top-class athletes (HMS and WCMS).

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