A novel imaging method (FIM-ID) reveals that myofibrillogenesis plays a major role in the mechanically induced growth of skeletal muscle.

An increase in mechanical loading, such as that which occurs during resistance exercise, induces radial growth of muscle fibers (i.e. an increase in cross-sectional area). Muscle fibers are largely composed of myofibrils, but whether radial growth is mediated by an increase in the size of the myofibrils (i.e. myofibril hypertrophy) and/or the number of myofibrils (i.e. myofibrillogenesis) is not known. Electron microscopy (EM) can provide images with the level of resolution that is needed to address this question, but the acquisition and subsequent analysis of EM images is a time- and cost-intensive process. To overcome this, we developed a novel method for visualizing myofibrils with a standard fluorescence microscope (fluorescence imaging of myofibrils with image deconvolution [FIM-ID]). Images from FIM-ID have a high degree of resolution and contrast, and these properties enabled us to develop pipelines for automated measurements of myofibril size and number. After extensively validating the automated measurements, we used both mouse and human models of increased mechanical loading to discover that the radial growth of muscle fibers is largely mediated by myofibrillogenesis. Collectively, the outcomes of this study offer insight into a fundamentally important topic in the field of muscle growth and provide future investigators with a time- and cost-effective means to study it.

Approximately 45% of human body mass is made of skeletal muscle. These muscles contract and relax to provide the mechanical forces needed for breathing, moving, keeping warm and performing many other essential processes. Both sedentary and active adults lose approximately 30-40% of this muscle mass by the age of 80, increasing their risk of disease, disability and death. As a result, there is much interest in developing therapies that can restore, maintain and increase muscle mass in older individuals. Muscles are made of multiple fibers that are in turn largely composed of smaller units known as myofibrils. Previous studies have shown that performing resistance training or other exercise that increases the mechanical loads placed on muscles stimulates muscle growth. This growth is largely due to increased girth of the existing muscle fibers. However, it remained unclear whether this was due to myofibrils growing in size, increasing in number, or a combination of both. To address this question, Jorgenson et al. developed a fluorescence imaging method called FIM-ID to count the number and measure the size of myofibrils within cross-sections of skeletal muscle. Using FIM-ID to study samples of mouse and human muscle fibers then revealed that increasing mechanical loads on muscles increased the number of myofibrils and this was largely responsible for muscle fiber growth. FIM-ID mostly relies on common laboratory instruments and free open-source software is used to count and measure the myofibrils. Jorgenson et al. hope that this will allow as many other researchers as possible to use FIM-ID to study myofibrils in the future. A better understanding of how the body controls the number of myofibrils may lead to the development of therapies that can mimic the effects of exercise on muscles to maintain or even increase muscle mass in human patients.

The Structural Adaptations That Mediate Disuse-Induced Atrophy of Skeletal Muscle.

The maintenance of skeletal muscle mass plays a fundamental role in health and issues associated with quality of life. Mechanical signals are one of the most potent regulators of muscle mass, with a decrease in mechanical loading leading to a decrease in muscle mass. This concept has been supported by a plethora of human- and animal-based studies over the past 100 years and has resulted in the commonly used term of 'disuse atrophy'. These same studies have also provided a great deal of insight into the structural adaptations that mediate disuse-induced atrophy. For instance, disuse results in radial atrophy of fascicles, and this is driven, at least in part, by radial atrophy of the muscle fibers. However, the ultrastructural adaptations that mediate these changes remain far from defined. Indeed, even the most basic questions, such as whether the radial atrophy of muscle fibers is driven by the radial atrophy of myofibrils and/or myofibril hypoplasia, have yet to be answered. In this review, we thoroughly summarize what is known about the macroscopic, microscopic, and ultrastructural adaptations that mediated disuse-induced atrophy and highlight some of the major gaps in knowledge that need to be filled.

A Novel Imaging Method (FIM-ID) Reveals that Myofibrillogenesis Plays a Major Role in the Mechanically Induced Growth of Skeletal Muscle.

An increase in mechanical loading, such as that which occurs during resistance exercise, induces radial growth of muscle fibers (i.e., an increase in cross-sectional area). Muscle fibers are largely composed of myofibrils, but whether radial growth is mediated by an increase in the size of the myofibrils (i.e., myofibril hypertrophy) and/or the number of myofibrils (i.e., myofibrillogenesis) is not known. Electron microscopy (EM) can provide images with the level of resolution that is needed to address this question, but the acquisition and subsequent analysis of EM images is a time- and cost-intensive process. To overcome this, we developed a novel method for visualizing myofibrils with a standard fluorescence microscope (FIM-ID). Images from FIM-ID have a high degree of resolution and contrast, and these properties enabled us to develop pipelines for automated measurements of myofibril size and number. After extensively validating the automated measurements, we used both mouse and human models of increased mechanical loading to discover that the radial growth of muscle fibers is largely mediated by myofibrillogenesis. Collectively, the outcomes of this study offer insight into a fundamentally important topic in the field of muscle growth and provide future investigators with a time- and cost-effective means to study it.

Protocol for quantifying the in vivo rate of protein degradation in mice using a pulse-chase technique.

The ability to measure the in vivo rate of protein degradation is a major limitation in numerous fields of biology. Here, we present a protocol for quantifying this rate in mice using a pulse-chase technique that utilizes an azide-bearing non-canonical amino acid called azidohomoalanine (AHA). We describe steps for using chow containing AHA to pulse-label the animal's proteome. We then detail the quantification of AHA-labeled proteins in whole-tissue lysates or histological sections using a copper-catalyzed azide-alkyne cycloaddition 'click' reaction. For complete details on the use and execution of this protocol, please refer to Steinert et al. (2023).1.

Shoulder, Trunk, and Hip Sagittal Plane Kinematics during Stand-Up Paddle Boarding.

Stand-up paddle boarding (SUP) as both a competitive and recreational sport has grown in popularity over the last decade. Better understanding paddling kinematics is beneficial for both injury prevention and informing coaching practices in this growing sport. The purpose of this study was to analyze sagittal plane kinematics during both standing and kneeling paddling postures commonly adopted by injury-free, recreational SUP participants. Eighteen recreational SUP participants (seven males/eleven females) were asked to complete a series of paddling tasks on a SUP ergometer in two postures, during which kinematic data were acquired. Sagittal plane kinematic data were analyzed for joint excursion, or range of motion used, while paddling on both sides of the body in each posture. Analysis of variance was used to compare joint excursions across tasks. There were no significant differences in hip or trunk sagittal plant excursion between postures. However, there was significantly greater sagittal plane excursion at the shoulder in the kneeling as compared to the standing posture with the shoulder opposite the paddling side demonstrating the greatest total excursion. These results help establish the parameters of the paddling technique currently in use among injury-free SUP participants and may be used in the future to inform coaching practices.

The role of satellite cell-derived TRIM28 in mechanical load- and injury-induced myogenesis.

Satellite cells are skeletal muscle stem cells that contribute to postnatal muscle growth, and they endow skeletal muscle with the ability to regenerate after a severe injury. Here we discovered that this myogenic potential of satellite cells requires a protein called tripartite motif-containing 28 (TRIM28). Unexpectedly, multiple lines of both in vitro and in vivo evidence revealed that the myogenic function of TRIM28 is not dependent on changes in the phosphorylation of its serine 473 residue. Moreover, the functions of TRIM28 were not mediated through the regulation of satellite cell proliferation or differentiation. Instead, our findings indicate that TRIM28 regulates the ability of satellite cells to progress through the process of fusion. Specifically, we discovered that TRIM28 controls the expression of a fusogenic protein called myomixer and concomitant fusion pore formation. Collectively, the outcomes of this study expose the framework of a novel regulatory pathway that is essential for myogenesis.

Strength, Cardiovascular Fitness, and Blood Lipid Measures in Law Enforcement Personnel After a 12-Week Health Promotion Program.

ABSTRACT: Hibbert, JE, Klawiter, DP, Schubert, MM, Nessler, JA, and Asakawa, DS. Strength, cardiovascular fitness, and blood lipid measures in law enforcement personnel after a 12-week health promotion program. J Strength Cond Res 36(11): 3105-3112, 2022-Law enforcement personnel often have high rates of cardiovascular disease and injury. Health promotion programs have been found to successfully encourage behavior change among law enforcement personnel, but these programs can often be intensive and expensive. Thus, the purpose of this study was to examine the efficacy of a health promotion program on body composition, metabolic health, muscle strength, and cardiovascular endurance in law enforcement personnel. Active duty officers from a local law enforcement agency were invited to participate in a 12-week health promotion program that included activity tracking and exercise and nutrition education. Eighteen subjects underwent measurements of body composition, V̇ o2 max through treadmill test, knee extensor strength, and blood lipids. An a priori alpha level for significance was set at 0.05, and comparisons were assessed using paired t -tests. Overall, subjects improved blood lipid levels evidenced by movement of mean values toward established healthy ranges. Although 8 subjects improved their V̇ o2 max (range: 1.3-30% change), there was large variability and no statistically significant differences in measures of V̇ o2 max (pre: 38.48 ± 5.86 ml·kg·min -1 ; post: 39.27 ± 5.26 ml·kg·min -1 ), body composition (pre: 26.52 ± 8.02% body fat; post: 26.44 ± 7.45% body fat), and strength normalized to body mass (isometric pre: 1.45 ± 0.45; post: 1.08 ± 0.36). Although no significant changes were noted, promising trends in these data suggest that health promotion programs with a modified focus may lead to positive changes in overall health.

Weight Pulling: A Novel Mouse Model of Human Progressive Resistance Exercise.

This study describes a mouse model of progressive resistance exercise that utilizes a full-body/multi-joint exercise (weight pulling) along with a training protocol that mimics a traditional human paradigm (three training sessions per week, ~8-12 repetitions per set, 2 min of rest between sets, approximately two maximal-intensity sets per session, last set taken to failure, and a progressive increase in loading that is based on the individual's performance). We demonstrate that weight pulling can induce an increase in the mass of numerous muscles throughout the body. The relative increase in muscle mass is similar to what has been observed in human studies, and is associated with the same type of long-term adaptations that occur in humans (e.g., fiber hypertrophy, myonuclear accretion, and, in some instances, a fast-to-slow transition in Type II fiber composition). Moreover, we demonstrate that weight pulling can induce the same type of acute responses that are thought to drive these long-term adaptations (e.g., the activation of signaling through mTORC1 and the induction of protein synthesis at 1 h post-exercise). Collectively, the results of this study indicate that weight pulling can serve as a highly translatable mouse model of progressive resistance exercise.

Measurement of intrinsic foot stiffness in minimally and traditionally shod runners using ultrasound elastography: A pilot study.

Running is an activity with a consistently high injury rate. Running footwear design that mimics barefoot running has been proposed to reduce injury rate by increasing the strength of foot structures. However, there is little evidence to support this. The purpose of the current study is to use shear wave ultrasound elastography to examine material properties (shear modulus) of intrinsic foot structures in experienced minimally and traditionally shod runners. It is hypothesized that minimalist runners will exhibit increased stiffness compared to controls demonstrating the strengthening of these structures. Eighteen healthy runners (8 minimalist and 10 traditionalist), running a minimum of 10 mi · wk-1, participated. Elastography scans were performed on the left foot of each participant. There is no apparent stiffening of foot structures associated with wearing minimalist shoes. Only the FHB tendon is different between shoe types and, contrary to the hypothesis, was stiffer in traditionalist compared to minimalist runners (257.26 ± 51.64 kPa vs 160.88 ± 27.79 kPa, respectively). A moderate positive (r = 0.7) relationship between training load and tendon stiffness suggests strengthening of tendon when running in traditional shoes. If running in minimalist shoes increases loading on these structures without resulting in stronger tissues, it is possible that minimalist footwear may increase injury risk.

Practice day may be unnecessary prior to testing knee extensor strength in young healthy adults.

A practice session is common prior to strength testing. However, the benefits of practice have not been previously reported. The purpose of this study was to determine the effect of a practice session on peak torque, mean torque and between trial variability across three test days. We hypothesized that peak and mean torque would be higher and less variable the second and third test days than the first. Twenty-five healthy, young participants completed 3 maximal voluntary isometric and isokinetic knee extensions on three separate days. No difference in isometric torque was found between days 1 and 2, but there was a significant decrease in isokinetic torque (8.45 Nm). There was a significant decrease in both mean isometric and isokinetic torque from day 1 to day 3 (12.67 and 13.59 Nm). Contrary to our hypothesis, no benefit from a practice session was found. Healthy, young adults are able to produce peak knee extensor torques on the first day of testing and do not demonstrate any benefit from additional testing. Thus, a practice day preceding isometric and isokinetic knee extensor strength testing may not be necessary when testing healthy, young participants, and may, in fact, negatively impact subsequent strength measurements.