How Is The Makeup Of Fibers Determined
Biol Sport. 2021 Jun; 38(2): 277–283.
Prediction of muscle fiber composition using multiple repetition testing
Elliott C.R. Hall,1 Evgeny A. Lysenko,2 Ekaterina A. Semenova,3 Oleg Five. Borisov,3, 4 Oleg Northward. Andryushchenko,v Liliya B. Andryushchenko,six Tatiana F. Vepkhvadze,ii Egor One thousand. Lednev,2 Piotr Zmijewski,7 Daniil 5. Popov,2 Edward 5. Generozov,3 and Ildus I. Ahmetov 
half dozen, 8, 9, 10
Elliott C.R. Hall
1Department of Sport and Exercise Sciences, Manchester Metropolitan University, Manchester, U.k.
Evgeny A. Lysenko
2Laboratory of Exercise Physiology, Constitute of Biomedical Issues of the Russian University of Sciences, Moscow, Russian federation
Ekaterina A. Semenova
3Department of Molecular Biology and Genetics, Federal Research and Clinical Center of Concrete-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia
Oleg V. Borisov
3Department of Molecular Biology and Genetics, Federal Enquiry and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia
fourInstitute for Genomic Statistics and Bioinformatics, Academy Infirmary Bonn, Bonn, Germany
Oleg N. Andryushchenko
vDepartment of Physical Teaching, Financial Academy under the Authorities of the Russian federation, Moscow, Russian federation
Liliya B. Andryushchenko
6Department of Concrete Education, Plekhanov Russian University of Economics, Moscow, Russia
Tatiana F. Vepkhvadze
2Laboratory of Exercise Physiology, Found of Biomedical Issues of the Russian Academy of Sciences, Moscow, Russian federation
Egor M. Lednev
2Laboratory of Practice Physiology, Establish of Biomedical Problems of the Russian Academy of Sciences, Moscow, Russia
Piotr Zmijewski
7Institute of Sport – National Research Establish, Warsaw, Poland
Daniil V. Popov
2Laboratory of Practise Physiology, Found of Biomedical Problems of the Russian Academy of Sciences, Moscow, Russia
Edward 5. Generozov
threeSection of Molecular Biology and Genetics, Federal Inquiry and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia
Ildus I. Ahmetov
6Section of Physical Education, Plekhanov Russian University of Economic science, Moscow, Russia
8Laboratory of Molecular Genetics, Kazan State Medical Academy, Kazan, Russia
9Sports Genetics Laboratory, St Petersburg Research Institute of Physical Culture, St Petersburg, Russia
10Research Institute for Sport and Exercise Sciences, Liverpool John Moores Academy, Liverpool, Great britain
Received 2020 Sep iii; Revisions requested 2020 Oct 4; Revised 2020 Oct three; Accustomed 2020 Oct 4.
Abstract
Direct determination of muscle fiber composition is invasive and expensive, with indirect methods also requiring specialist resource and expertise. Performing resistance exercises at eighty% 1RM is suggested as a means of indirectly estimating muscle fiber composition, though this hypothesis has never been validated against a direct method. The aim of the written report was to investigate the relationship between the number of completed repetitions at fourscore% 1RM of back squat practice and muscle fiber limerick. Xxx recreationally active participants' (10 females, 20 males) 1RM back squat load was determined, earlier the number of sequent repetitions at 80% 1RM was recorded. The relationship between the number of repetitions and the percent of fast-twitch fibers from vastus lateralis was investigated. The number of completed repetitions ranged from 5 to 15 and was contained of sex, age, 1RM, training frequency, training type, preparation experience, BMI or muscle fiber cross-exclusive area. The percentage of fast-twitch musculus fibers was inversely correlated with the number of repetitions completed (r = –0.38, P = 0.039). Participants achieving five to viii repetitions (northward = x) had significantly more fast-twitch muscle fibers (57.5 ± 9.5 vs 44.4 ± 11.ix%, P = 0.013) than those achieving eleven–fifteen repetitions (n = 11). The remaining participants achieved ix or 10 repetitions (north = 9) and on average had equal proportion of fast- and slow-twitch musculus fibers. In decision, the number of completed repetitions at 80% of 1RM is moderately correlated with musculus cobweb composition.
Keywords: Cobweb type, Strength training, Vastus lateralis, Exercise prescription, Endurance
INTRODUCTION
The variable individual response to practise training [1] is likely to exist influenced by the heterogeneity of musculus cobweb limerick [2, 3]. Oxidative type I (slow-twitch) musculus fibers are predominantly suited to aerobic activities and repetitive submaximal contractions, whilst type II (fast-twitch, subdivided into oxidative type IIa and glycolytic type IIx) fibers accept greater cross-sectional surface area and faster shortening velocity than type I fibers [4], and are preferentially recruited during anaerobic activities necessitating strength and power [five]. Whilst appropriate programme design is important in attaining the desired response to sustained training, variable inter-private responses exist and are probable to be influenced by the muscle fiber composition of each individual. Appropriately, information concerning the individual variability in muscle cobweb composition could help coaches and practitioners when designing individualised training programmes tailored toward specific physiological adaptations [vi].
The vastus lateralis (VL) musculus is key contributor to athletic movements such as running and jumping [7]. However, the proportion of cobweb types in the VL varies considerably, ranging between fifteen–85% blazon I, 5–77% type IIa, and 0–44% type IIx [eight–10]. The golden-standard to directly determine muscle fiber composition is the musculus biopsy, with the VL most usually sampled due to its mixed fiber limerick and accessibility [11]. Subsequently, the fiber composition of biopsies is determined using methods such every bit immunohistochemical analysis, histochemical staining for myosin ATPase, gel electrophoresis and mass-spectrometry [12–14], with transcriptomic analysis of the MYH1, MYH2 and MYH7 genes recently used to determine type IIx, type IIa and type I muscle fibers, respectively [fifteen]. Still, collecting muscle biopsies is invasive, and subsequent analyses are expensive to perform, limiting accessibility to direct analytical methods.
Indirect methods to judge muscle fiber composition offer culling approaches that tin be validated against directly methods. Tensiomyography (TMG) evaluates the morphofunctional potential of a muscle according to the radial enlargement of the muscle abdomen in response to electrical stimulation [16], with time to muscle contraction well correlated with slow-twitch muscle fiber proportion [17]. With fast-twitch fibers containing twice the carnosine content of irksome-twitch fibers, magnetic resonance spectroscopy (MRS) of carnosine content is also indicative of fiber type [18]. This method was recently used to predict recovery from Wingate cycling, with considerable differences in knee extensor strength recovery between participants with predominantly irksome-twitch fibers and participants with predominantly fast-twitch fibers [19]. Musculus fiber composition can also be estimated according to the vibrational properties of contracting skeletal musculus by mechanomyography (MMG), which demonstrates fourscore% accuracy in predicting VL fiber limerick [20]. In addition to morphological measures, skeletal musculus phenotypes are associated with mutual genetic variants, suggesting that testing individuals for known genetic variants may assist the estimation of muscle fiber composition. Specific genes associated with muscle fiber blazon include ACE, ACTN3, AGTR2, CBLN2, CPNE5, HIF1A, FTO, PPARA, SPEG, TGFA, and VEGFR2 [21–33], with additional variants likely to exist. However, genomic inquiry requires specialist cognition and resources, farther demonstrating the potential value of simple and price-effective methods to indirectly approximate muscle fiber composition.
Inter-individual differences are axiomatic in the number of completed repetitions at different percentages of one repetition maximum (1RM), with heterogeneity in muscle fiber limerick suggested as an underlying mechanism [34]. Appropriately, the number of repetitions achieved at specific percentages of 1RM has been proposed as a practical arroyo to estimate muscle fibre composition [35]. However, despite the clan of exertion at specific percentages of 1RM with the variability in repetitions performed [36], at that place are no published studies reporting significant correlations betwixt direct estimates of muscle fiber composition and multiple repetition testing. In i study by Terzis et al. [37], leg press machine testing was used to determine the association of 70% and 85% 1RM repetition tests with musculus fibre composition, with participants possessing greater proportions of boring-twitch muscle fibres completing more repetitions. However, this result did non reach statistical significance, potentially due to a limited sample size (n = 12). With admission to a leg printing motorcar not e'er possible, back squat exercise using an Olympic barbell offers a viable alternative for practice testing. Furthermore, there is also a need to study participants of both genders and to exercise then with a sample size greater than previous investigations. Therefore, the aim of this study was to determine the interrelationship between musculus cobweb composition of the vastus lateralis of thirty participants (10 females and xx males) and the total number of consecutive repetitions achieved at 80% of 1RM using back squat exercise. It was hypothesised that the number of repetitions completed would be inversely correlated with the proportion of fast-twitch muscle fibers and would exist independent of gender.
MATERIALS AND METHODS
Subjects
Thirty recreationally active participants (10 females, age 26.0 ± 7.3; 20 males; historic period 32.2 ± 7.6) participated in the written report. Participant characteristics are shown in Table 1. Participants' training parameters were assessed using a standardised questionnaire, with participants classified according to preparation frequency every bit moderately active (iii–iv training sessions per week; north = 11), highly active (v–seven training sessions per week; n = eleven) and extremely active (ane–ii grooming sessions per day; n = viii). Participants' training backgrounds were classified equally either resistance type (three females, 10 males) or mixed type (aerobic + resistance; seven females, x males), with training experience expressed every bit the total number training years. The procedures adopted in this written report were conducted ethically according to the Declaration of Helsinki for research involving man subjects. Ethical approval was obtained from the Ethics Committee of the Physiological Department of the Russian National Committee for Biological Ideals in Russia and the Ethics Commission of the Federal Research and Clinical Centre of Concrete-chemical Medicine of the Federal Medical and Biological Agency of Russia. Written informed consent was obtained from all participants prior to participation.
Tabular array 1
Characteristics and practise testing results of study participants
| Characteristics | Female person (n = 10) | Male (due north = 20) | P |
|---|---|---|---|
| Mean ± SD | Mean ± SD | ||
| Age, years | 26.0 ± 7.3 | 32.2 ± 7.6 | 0.024 |
| Training experience, years | 10.4 ± 7.0 | eleven.viii ± 7.8 | 0.725 |
| Superlative, cm | 169.0 ± v.2 | 179.iv ± v.viii | 0.0003 |
| Weight, kg | 57.3 ± six.2 | 80.1 ± 13.8 | < 0.0001 |
| BMI, kg/1000two | 20.0 ± one.2 | 24.nine ± iv.3 | 0.0005 |
| Proportion of fast-twitch MF, % | 48.4 ± 10.9 | 52.2 ± 15.iii | 0.619 |
| Proportion of ho-hum-twitch MF, % | 56.three ± ix.viii | 51.1 ± 15.4 | 0.397 |
| CSA of fast-twitch MF, μm2 | 4173 ± 1488 | 6625 ± 2530 | 0.004 |
| CSA of dull-twitch MF, μm2 | 4662 ± 1013 | 5333 ± 1743 | 0.176 |
| 1RM squat (kg) | 70.five ± 31.3 | 155.2 ± 62.7 | 0.0005 |
| Completed repetitions at lxxx% 1RM | 9.8 ± 2.7 | 10.3 ± 2.viii | 0.659 |
Procedures
Back squat testing
All participants were familiar with dorsum squat exercise with a minimum of 2 years experience. Participants performed a 1RM protocol to decide the maximal load they could lift for the back squat with correct technique using a standard 20 kg Olympic barbell. Briefly, participants performed a warm-upwardly with a self-selected load that allowed them to complete a minimum of eight repetitions. For each 1RM attempt, participants squatted (descending and ascending for ii s and i southward, respectively) with the bar placed beyond the posterior deltoids and until the tops of their thighs were parallel to the ground [38]. One repetition maximum was determined according to guidelines published by the National Strength and Conditioning Association [38]. After successfully achieving their 1RM load, participants rested passively for 15 minutes. Subsequently resting, participants were instructed to perform every bit many consecutive repetitions at 80% of their individualised 1RM load in one continuous endeavour, adhering to the same criteria for descent, rising and bar positioning as during 1RM attempts. All attempts were vocally encouraged, directed and supervised by the exam administrator who was qualified in exercise prescription. A load of fourscore% 1RM was selected because this load is recommended for increasing muscle strength and achieving musculus hypertrophy [39], meaning this load is commonly prescribed by practitioners working with the athletes and general population, and considering lxxx% 1RM was originally proposed equally the most advisable load to indirectly estimate musculus fiber composition [35].
Evaluation of musculus fiber composition and cross-exclusive area
Percutaneous musculus biopsies were collected from the vastus lateralis of the left leg using the modified Bergström needle procedure with aspiration nether local anesthesia with 2% lidocaine solution. Samples were immedately frozen in liquid nitrogen and stored at -80°C. For analysis, serial cross-sections (7 μm) were obtained from frozen samples using an ultratom (Leica Microsystems, Federal republic of germany). Sections were thaw-mounted on polysine glass slides, maintained at room temperature (RT) for fifteen min and incubated in PBS (3 v min). The sections were and so incubated at RT in principal antibodies confronting slow or fast isoforms of the myosin heavy chains (M8421, i:5000; M4276; 1:600, respectively; Sigma-Aldrich, U.s.) for ane h and incubated in PBS (iii × 5 min). Adjacent, the sections were incubated at RT in secondary antibodies conjugated with FITC (F0257; 1:100; Sigma-Aldrich) for i h. The antibodies were removed and the sections washed in PBS (iii × 5 min), placed in mounting media and covered with a cover slip. Images were captured by fluorescent microscope (Eclipse Ti-U, Nikon, Japan). All analyzed images contained 318 23 fibers (Fig. 1). The ratio of the number of stained fibers to the total fiber number was calculated. Fibers stained in serial sections with antibodies against irksome and fast isoforms were considered hybrid fibers. The cantankerous-sectional expanse (CSA) of fast- and ho-hum-twitch musculus fibers was evaluated using ImageJ software (NIH, Us).
Microphotographs of the labelled muscle sections.
Statistical analyses
Statistical analyses were conducted using GraphPad InStat (GraphPad Software, Inc., United states of america) software. Differences in traits betwixt different groups were analysed using Mann-Whitney test. Relationships between participant characteristics, exercise testing performance and muscle fiber measurements was tested using Spearman'southward (non-parametric) correlation. Multiple regression analysis was used to detect independent associations between the number of repetitions achieved at fourscore% of 1RM and different characteristics. All information are presented equally mean (SD). P values < 0.05 were considered statistically significant.
RESULTS
Participants exercise operation during 1RM and eighty% 1RM testing, besides as muscle fiber characteristics are presented separately for females and males in Table 1. The number of repetitions accomplished at 80% of 1RM ranged from five to 15 and was contained of sexual activity, age, 1RM, training frequency, type and experience, BMI and CSA of muscle fibers (all P > 0.05). The per centum of fast-twitch muscle fibers was inversely correlated with the number of repetitions completed (r = –0.38, P = 0.039, Fig. ii). Participants who completed 5 to 8 repetitions (n = 10; iv females and half dozen males) had significantly more fast-twitch muscle fibers (57.5 ± ix.5 vs 44.4 ± eleven.9%, P = 0.013) than participants who completed eleven to 15 repetitions (n = 11; four females and vii males). The remaining participants completed nine or x repetitions (due north = ix; 2 females and seven males) and on boilerplate had equal proportion of fast (51.six ± 17.five%) and tedious-twitch (50.8 ± sixteen.9%) muscle fibers.
Relationship between muscle fiber limerick and the number of completed repetitions at 80% 1RM. Shaded areas represent 95% confidence intervals.
DISCUSSION
The aim of the present study was to determine whether musculus cobweb composition of the VL is associated with the number of consecutive back squat repetitions completed at lxxx% 1RM. The principal finding was a moderate inverse correlation between the per centum of fasttwitch musculus fibers and the number of repetitions completed, which was unrelated to sex activity, age, 1RM, training frequency, preparation type, training age, BMI or fast-twitch muscle cobweb CSA. This study is the first to demonstrate concordance between the use of an 80% 1RM load during back squat exercise and the muscle cobweb composition of VL biopsies, signifying the potential of this approach to indirectly guess VL musculus fiber limerick.
In the present study, participants achieving v to 8 repetitions had a higher percentage of fast-twitch fibers than those achieving xi to fifteen repetitions (57.5 vs 44.3%). Those who completed 9 to ten repetitions exhibited an equal proportion of each fiber blazon. Our hypothesis was centred on the theory that loads of 80% 1RM load could exist used to gauge the muscle fiber limerick of untrained individuals, which had not been previously tested using back squat exercise. It was suggested that participants completing more than 12 repetitions would possess more than 50% ho-hum-twitch fibers, individuals completing fewer than 7 repetitions would possess more than 50% fast-twitch fibers, and that those completing between 7 and 12 repetitions would exhibit an equal proportion of each cobweb type [35]. To our knowledge, the present report is the starting time to find a pregnant interrelationship between muscle cobweb composition of the VL and the variability in the number of back squat repetitions completed at lxxx% 1RM.
Our results are supported by previous literature where participants estimated to posses more than fast-twitch musculus fibers completed fewer repetitions at an individualised load than those estimated to possess more slow-twitch fibers [36]. Comparing with that report is limited by the recruitment of only female participants, the investigation of a 70% 1RM load, and the fact that testing was performed on an isokinetic dynamometer. Still, participants in the nowadays written report completed a like range of repetitions (5 to 15) to the previous study (7 to 15), with a slightly lower hateful number of repetitions, which may reverberate the current participants lifting a relatively greater load. Whilst the present study investigated this relationship using conventional resistance practice and muscle biopsies, the previous study associated isokinetic dynamometry measures with an indirect estimate of muscle cobweb composition derived from a regression equation [40]. The equation described incorporates relative torque after 55 contractions of a fatigue test with power output at an angular velocity of 280°s-1 normalised to fat-free mass of the thigh. Together, these variables explained 51.eight% of the variance in the percentage of VL fast-twitch fibers. Whilst this clan of skeletal muscle phenotypes with musculus fiber composition is of involvement, the practical use of this equation is limited past the requirement to access an isokinetic dynamometer and to accurately quantify fatty-gratis-mass. In contrast, the present study demonstrates the relationship between directly measured VL cobweb composition and a universally recognised resistance do, with individualised loads derived using a reliable assessment of 1RM [41]. Whilst further studies in larger cohorts are required to back up our findings, the nowadays report highlights the potential of this arroyo as an accessible and cost-effective alternative for estimating muscle fiber limerick.
No relationship was observed between participants' grooming groundwork and muscle cobweb composition in the present report. Previously, endurance athletes completed more than repetitions of leg printing exercise at 70% and 80% 1RM compared to strength athletes, with no difference at 90% [42], whilst non-athletes accustomed to endurance training accomplished more repetitions at 80% 1RM of half-squat exercise than those accepted to strength preparation [43]. These results are of importance because endurance trained individuals typically possess a greater percentage of ho-hum-twitch fibers [3] demonstrating increased lactate buffering chapters [44], fiber capillarisation [45] and mitochondrial content [46], i.e. the muscles of these individuals take greater oxidative capacity and fatigue resistance than their strength trained counterparts [11]. Ho-hum-twitch muscle fibers are suited to repetitive and submaximal contractile activity, which might explain why participants with a greater proportion of slow-twitch fibers completed a greater number of repetitions at eighty% 1RM in the present study. It is possible that discrepancies between this and previous studies reverberate the exercises studied, or the different athletic backgrounds of participants. Given that the number of repetitions completed at 80% of 1RM was related to cobweb composition but was independent of age, 1RM, training frequency, grooming type, training experience, BMI and fast-twitch musculus cobweb CSA, it is plausible to suggest that this trait is partly heritable, with further investigation required to examination this hypothesis.
Directly decision of muscle cobweb composition is invasive and expensive, and therefore unfavourable to field settings and the general population. Many indirect methods require specialist resources and have their own limitations. For example, TMG requires a highly trained operator to minimise measurement fault [47], the detection of carnosine content by MRS is heavily influenced past dietary factors [48], and crosstalk from neighbouring muscles during MMG affects the myographic signal from the investigated region [49]. Appropriately, there is a need for an indirect and accessible method to estimate fiber composition that can be executed by practise professionals using standard equipment. The nowadays study is the showtime to experimentally validate a previous hypothesis [35] using back squat exercise and to demonstrate that the number of repetitions achieved at 80% 1RM is associated with muscle fiber composition. Our findings highlight the potential to develop a non-invasive and cost-constructive approach for estimating muscle fiber composition which, following farther validation, could aid the blueprint of training programmes for improving the strength or endurance chapters of skeletal muscle. With fast-twitch fibers preferentially damaged post-obit fast and forceful contractions [50], knowledge of fiber composition may likewise assist recovery from strenuous training. Appropriately, this field test could help practice professionals identify individuals who may be more susceptible to exercise induced musculus harm, and who may require extended residual or additional recovery strategies post-obit strenuous training. At present, our findings are restricted to males and females aged 18 to twoscore, with replication required before this test tin be applicable to other ages, such as youths and the elderly. Furthermore, because the reliability of maximal back squat testing increases after half-dozen–12 months of resistance grooming [51], the use of this test may be inappropriate for less experienced individuals.
The advantages of the present study include the direct decision of VL muscle fiber composition, a common and reliable method for 1RM testing [41], and the potential practical application of these findings for individuals without access to the facilities and expertise required to directly determine fiber limerick. Even so, this study also has limitations. Firstly, we recognise that a greater sample size could amend the statistical power of the results. Secondly, our findings are limited to the musculus fiber limerick of the VL, and due to the heterogeneity of fiber composition within skeletal muscles of the homo body, the validity of this method is limited to the VL. We also recognise that data regarding participants' endurance exercise capacity and genetic profile was not included in the present study, and that including such data may amend the prediction of muscle fiber composition in future studies, in addition to investigating a series of external loads. Finally, we recognise that validation of this arroyo in untrained populations and in aristocracy athletes is required for this approach to be used with sedentary individuals and within highlevel sport, for example in weightlifting and powerlifting where the back squat is a key exercise.
CONCLUSIONS
The present study is the kickoff to demonstrate the interrelationship between back squat performance at eighty% 1RM and muscle fiber limerick. We propose that completing viii or fewer repetitions at 80% 1RM is indicative that participants possess more than than 50% fast-twitch fibers in their VL, that completing 11 or more than repetitions at 80% 1RM is indicative that participants possess more than fifty% slow-twitch fibers in their VL, and that completing 9 or x repetitions is indicative of participants possessing an equal distribution of each fiber type. Though farther validation is required, the present study demonstrates the association of musculus fiber type with dorsum squat functioning and highlights potential to develop a non-invasive and cost-effective arroyo to approximate muscle fiber limerick.
Conflict of interest
The authors declare no conflict of interest.
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Articles from Biology of Sport are provided here courtesy of Institute of Sport
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8139349/
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