CN103160607A - Biological method for predicating sensitivity potentials of excellent ice-snow sportsmen - Google Patents
Biological method for predicating sensitivity potentials of excellent ice-snow sportsmen Download PDFInfo
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Abstract
The invention discloses a molecular biology method for predicating the muscle sensitivity potentials of excellent ice-snow sportsmen, which can be used for inquiring the relationship between the R577X polymorphisms of ACTN3 (alpha actin in, alpha-actin-3) genes and the muscle sensitivity potentials of the excellent ice-snow sportsmen. With the continuous development of molecular biology technology, applications for analysis for genes related to movements in sports science become a research direction with a great prospect; polymorphic analysis for ACTN3 is mainly researched at present; the existence of ACTN3 is beneficial to the development of the muscle sensitivity skills of speed-type and strength-type movements, and the lack of ACTN3 may lower the level of the movement skills; therefore, the occurrence frequency of the XX genotypes of ACTN3 (the lack of ACTN3 proteins) in the excellent speed-type and strength-type sportsmen is low. The molecular biology method disclosed by the invention belongs to the fields of the interdisciplines of sports science, human resource methodology, cytobiology and molecular biology.
Description
Technical field
The present invention relates to a kind of molecular biology method of predicting the sensitive potential of outstanding winter sports person's muscle, be exactly specifically to pass through ACTN3 (a1pha act in, α-actin-3) cognation of gene R577X polymorphism and outstanding winter sports person's motor capacity is studied, thereby outstanding winter sports person's muscle susceptibility potential is predicted.This invention belongs to the interdisciplinary field of Basic Life science, mankind's inheritance resources, molecular biology and sports science.
Background technology
Sensitivity refers to complete correctly, rapidly, rationally and neatly the ability of not familiar, new action, namely grasps rapidly the ability of action under change condition.In the antagonism motion of fierceness, along with the variation of mode of motion, respective change all can occur in the character of action, intensity, and pallium must promptly be made situation about changing and be analyzed judgement, and decides on the spot.The contraction speed of muscle, the handiness of Neuromuscular Conduction are the basic substances of sensitive fitness.So movable by detecting neuromuscular, measurable people gets muscular movement handiness and susceptibility.
The research data of synthesise various affects flexibly or the factor of susceptibility comprises following content: (1) speed (speed of response, responsiveness); (2) strength (explosive strength, dorsal muscles strength, leg muscular strength amount); (3) Harmony; (4) pliable and tough (handiness of waist marrow, knee, ankle handiness); (5) endurance (stamina of speed, general endurance); (6) state of the art (what and skill level technical movements grasp); (7) types of nerve; (8) psychological condition; (9) orientation property accurately on room and time; (10) action nerve process handiness; (11) judgement fast and accurately; (12) range and the stability noted; (13) balance ability; (14) body shape; (15) age and sex; (16) fatigue.
The Molecular Biology Mechanism of motor capacity receives much concern, and the Study on gene polymorphism that wherein motor capacity is relevant has become one of focus.In the existing research of motor capacity candidate gene, α-actin-3 (ACTN3) gene R577X polymorphism is to be only second to ACE (Angiotensin-converting enzyme) gene I/D (kininase II gene insertion/deletion type) polymorphism, and research quantity comes the polymorphic site of the 2nd.The ACTN3 gene is positioned at 11q13-q14, exist C → T polymorphic on its 16th exon, make the codon CGA (coding arginine R) of coding the 577th amino acids become TGA and (be termination signal, proteins encoded not), thereby cause the disappearance of ACTN3, be called ACTN3 gene R577X polymorphism (rs1815739, or be called the C1747T polymorphism).In the report about the ACTN3 gene, Yang is to the athletic research discovery of Australian elite male and female Velocity-force type, 577R allelotrope is significantly higher than control group in the frequency of Velocity-force group, and the 577R allelotrope of prompting ACTN3 gene is advantage allelotrope for strength quality and speed quality.
The mankind have the skeletal muscle fibre of two types.A kind of is fastfiber, and it does not need oxygen, directly with carbohydrate as the energy, main needing to participate in limit strength and quick response motion, such as dash.Another kind is slowfiber, mainly needing to participate in the motion of endurance in the mode of aerobic, such as Phosphothion.Fastfiber can be made α actinin (ACTN3), and it has embodied the ability of the quick strength of body generation.If the ACTN3 gene pleiomorphism to outstanding winter sports person is analyzed, grasp athletic genetic dominance; In athletic training and competition, the sportsmen of different genotype is placed on different positions, give full play to its motion advantage, rationally utilize the motion resource, will be conducive to improve the competitiveness of group project.
ACTN3 is the structural protein of fast muscle fiber Z line in skeletal muscle, and is crosslinked mutually with the Actin muscle in thin filament, keeps myofibrillar ordered arrangement and regulates myofibrillar contraction.Although do not get rid of other factors to the regulating effect of muscle fiber types.Be beneficial to the development of Velocity-force type technical performance due to having of ACTN3, the shortage of ACTN3 will reduce the level of technical performance, and it is less that the frequency of ACTN3XX genotype (ACTN3 hypoproteinosis) appears in inference in outstanding Velocity-force type sportsmen thus.
The most basic, the most important mode that neurone is completed its function is to carry out intercellular information transmission by cynapse.Information transmission from the motor neuron to myocyte is mainly completed by neuromuscular junction.Neuromuscular junction is the Typical Representative of directed chemical synapse, its important constitutional features is the postsynaptic membrane of eggcase, it is myocyte's telolemma, it can preferentially identify neurogenic signal, it is obviously different from extrasynaptic plasma membrane at ultrastructure and molecular level: at first, postsynaptic membrane abnormal clusters collection (cluster) a large amount of acetylcholine receptor (acetycholine receptor, AChR) structure, and density is up to 1000 acceptors/μ m
2, extrasynaptic plasma membrane AChRs density is less than 10 acceptors/μ m
2, this distribution of AChRs is essential to Neuromuscular communication effectively; Secondly, postsynaptic membrane has minute submounts of a uniqueness, and the specialization molecule of postsynaptic membrane is anchored on basilar membrane and cytoskeleton, and postsynaptic differentiation is had important effect.Closely studies show that, after the presynaptic, cell all participates in the formation of AChR bunch of collection and keeps.AChRs bunch of collection and 3 kinds of molecules are closely related, be agrin (Agrin), the peculiar receptor tyrosine kinase of flesh (muscle-specific receptor tyrosine kinase, MuSK) and postsynaptic membrane acceptor Rapsyn (acetylcholine receptor-associated protein at synapse, Rapsyn); Agrin induces AChR at bunch collection of telolemma, and MuSK is one of important component of Agrin signal transduction acceptor complex body, and Rapsyn participates in its effect mechanism.
Acetylcholine receptor can be identified and in conjunction with neurotransmitter acetylcholine.In case both combinations, the ionic channel of acetylcholine receptor coupling is opened, and increases the chance that positively charged ion enters adipose membrane.Like this, ionic channel is opened, and the aqueous solution on cell phospholipid bilayer two sides has again a new ion concentration gradient.The outer solution of born of the same parents has high potassium concentration (outflow), and in born of the same parents, solution has high Na ion concentration (inflow).The change of concentration balance causes the film depolarize.The depolarize of film causes again the specific physiological response of neurocyte.Along with the cytolemma degree of depolarization increases, can cause action potential, action potential propagates into myofiber by the muscle neuron membrane.The propagation of action potential impels the release of the intermembranous calcium ion of muscle cell simultaneously, finally causes the contraction of flesh ball fiber.All acetylcholine receptor characteristics relevant with biological activity are all causing when neurotransmitter acetylcholine is combined, and acetylcholine receptor is after vagusstoff is combined, and receptor conformation will change.
Summary of the invention
The present invention can predict outstanding winter sports person's muscle sensitivity potential.
The purpose of this invention is to provide a kind of molecular biology method of predicting outstanding winter sports person's muscle sensitivity potential.
Forecasting Methodology provided by the present invention is the cognation research of ACTN3 gene R577X polymorphism and motor capacity.The PCR product is carried out the endonuclease reaction of restriction enzyme Dde I (TOYOBO), cut the quantity of after product band according to enzyme and come the identified gene type: wild-type (RR type) will produce 2 fragments (392bp, 168bp).Mutant (XX type) produces 3 fragments (295,168,97bp).If heterozygote (RX type), four fragments (392,295,168,97bp) just appear.Compare with common control group, the RR genotype (47%, 31%) of winter sports person's group and the frequency of occurrences of R allelotrope (66%, 56%) are higher.
Embodiment
The below carries out ACTN3 gene R577X polymorphism analysis as example take outstanding winter sports person and common artificial experimental subjects, describes concrete enforcement of the present invention in detail.
Embodiment: ACTN3 gene R577X polymorphism analysis
Research object
Research object is mainly divided 2 groups, outstanding winter sports person's group and the general population's group, and the general population organizes main selection people from this area suitable with this elite's age, carries out in strict accordance with the principle of stochastic sampling, all without professional training history.
(1) DNA extraction
Use the buccal swab genome DNA extracting reagent kit (centrifugal column type) that sky, Beijing root biochemical technology company limited provides to extract oral cavity DNA without wound, operation to specifications.
(2) gene amplification
Upstream primer: 5 '-AGGGGATGGATAGGATGACAGG-3 '
Downstream primer: 5 '-ATCCCACGTGGAGTCTGTGG-3 '
The PCR reaction conditions is to unwind: 94 ℃, and 5 minutes; Annealing: 94 ℃, 30 seconds; 72 ℃, 45 seconds; 58 ℃, 30 seconds; Extend: 72 ℃, 5 minutes; Cycle index: 35 times.
Pcr amplification product 5 μ l, 1.5% sepharose constant voltage 80V electrophoresis 30-40 minute, EB dyeing 10 minutes is put and is detected pcr amplification product and specificity thereof on the gel imaging instrument, and product length is 560bp.
(3) endonuclease reaction
The PCR product carries out the endonuclease reaction of restriction enzyme Dde I (TOYOBO), detect on the gel imaging instrument after electrophoresis, cut the quantity of after product band according to enzyme and come the identified gene type: wild-type (RR type) will produce 2 fragments (392bp, 168bp).Mutant (XX type) produces 3 fragments (295,168,97bp).If heterozygote (RX type), four fragments (392,295,168,97bp) just appear.
(4) statistical procedures of data
Selected general population's data must meet mendelian inheritance, namely reaches Hardy-Weinberg balance (P＞0.05).Then enumeration data is compared analysis (X
2Check), P＜0.05 expression difference has statistical significance.
(5) qualification result
General population's ACTN3 genotype frequency is: RR type 0.3444, RX type 0.5000, XX type 0.1556 meet Hardy-Weinberg balance (X
2=0.1232, P=0.7256), show that the data obtained is representative to overall crowd.
Outstanding winter sports person's group has significant difference (P=0.031, P＜0.05) with the genotype distribution of the ACTN3 gene R577X polymorphic site of common control group, and allele distributions also has significant difference (P=0.022, P＜0.05).Compare with common control group, the RR genotype (47%, 31%) of winter sports person's group and the frequency of occurrences of R allelotrope (66%, 56%) are obviously higher.
ACTN3 has a development that is beneficial to Velocity-force type technical performance, the shortage of ACTN3 will reduce the level of technical performance, the frequency that occurs thus ACTN3XX genotype (ACTN3 hypoproteinosis) in outstanding winter sports person is less, and ACTN3RR type frequency is higher.
Claims (3)
1. method of probing into ACTN3 (alpha act in, α-actin-3) gene R577X polymorphism and outstanding winter sports person's muscle sensitivity potential relation.
2. method according to claim 1, the innovative point of this method is the dependency by quantitative evaluation ACTN3 gene R577X polymorphism and Velocity-force quality, thereby can be with ACTN3 gene R577X polymorphism as one of candidate locus of Velocity-force type project sportsmen muscle susceptibility gene selection.
3. according to claim 1 with 2 is described: having of ACTN3 is beneficial to the sensitive skill development of Velocity-force type motion muscle, the shortage of ACTN3 will reduce the level of technical performance, and the frequency that therefore occurs ACTN3XX genotype (ACTN3 hypoproteinosis) in outstanding Velocity-force type sportsmen is less.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115521986A (en) * | 2022-09-19 | 2022-12-27 | 大连理工大学 | Human physical ability evaluation method based on biological multigroup knowledge |
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| KR102582848B1 (en) * | 2022-11-09 | 2023-09-27 | 삼육대학교산학협력단 | Composition with genetic properties beneficial to power sports |
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Non-Patent Citations (3)
| Title |
|---|
| DRUZHEVSKAYA AM ET AL: "Association of ACTN3 R577X polymorphism with power athlete status in Russians", 《EUROPEAN JOURNAL OF APPLIED PHYSIOLOGY》 * |
| 杨贤罡 等: "ACTN3基因R577X多态性与运动能力的关联性研究：Meta分析", 《体育科学》 * |
| 贺兰湘 等: "ACTN3基因多态性与运动技能的发展", 《成都体育学院学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115521986A (en) * | 2022-09-19 | 2022-12-27 | 大连理工大学 | Human physical ability evaluation method based on biological multigroup knowledge |
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