Allometry: revealing evolution's engineering principles.

An animal's body size impacts many aspects of its structure and function (Y); insights that only become apparent when viewed over several orders of magnitude of body mass (M) and expressed allometrically in the form of power law equations (Y=aMb). The resulting relationships are apparent clusters of similar exponents (b) revealing emergent 'patterns of design' that shed light on the universal principles of structure and function. Basic physical principles of surface area, volume and heat exchange apply to all objects, including animals, and many consequences must be attributed to these fundamental properties. Starting with Galileo's description of the shapes of bones in the 16th century and extending to 19th century explanations of heat production and loss by Sarrus and Rameaux, allometric patterns have provided numerous biological insights. Here, we examine several of these insights and explore how the selective pressures and scaling may differ when comparing animals in a vegetative (basal) state and those utilizing their maximum metabolic capacities. It seems that the selective pressures under those two conditions differ. We caution that allometric patterns invite explanations that lack supporting data or may be dismissed because there is hesitation among biologists to make comparisons lacking phylogenetic support. We argue that emergent allometric patterns have inherent value and continue to be the fodder for many fruitful hypotheses.

N2A Titin: Signaling Hub and Mechanical Switch in Skeletal Muscle.

Since its belated discovery, our understanding of the giant protein titin has grown exponentially from its humble beginning as a sarcomeric scaffold to recent recognition of its critical mechanical and signaling functions in active muscle. One uniquely useful model to unravel titin's functions, muscular dystrophy with myositis (mdm), arose spontaneously in mice as a transposon-like LINE repeat insertion that results in a small deletion in the N2A region of titin. This small deletion profoundly affects hypertrophic signaling and muscle mechanics, thereby providing insights into the function of this specific region and the consequences of its dysfunction. The impact of this mutation is profound, affecting diverse aspects of the phenotype including muscle mechanics, developmental hypertrophy, and thermoregulation. In this review, we explore accumulating evidence that points to the N2A region of titin as a dynamic "switch" that is critical for both mechanical and signaling functions in skeletal muscle. Calcium-dependent binding of N2A titin to actin filaments triggers a cascade of changes in titin that affect mechanical properties such as elastic energy storage and return, as well as hypertrophic signaling. The mdm phenotype also points to the existence of as yet unidentified signaling pathways for muscle hypertrophy and thermoregulation, likely involving titin's PEVK region as well as the N2A signalosome.

Basic science and clinical use of eccentric contractions: History and uncertainties.

The peculiar attributes of muscles that are stretched when active have been noted for nearly a century. Understandably, the focus of muscle physiology has been primarily on shortening and isometric contractions, as eloquently revealed by A.V. Hill and subsequently by his students. When the sliding filament theory was introduced by A.F. Huxley and H.E. Huxley, it was a relatively simple task to link Hill's mechanical observations to the actions of the cross bridges during these shortening and isometric contractions. In contrast, lengthening or eccentric contractions have remained somewhat enigmatic. Dismissed as necessarily causing muscle damage, eccentric contractions have been much more difficult to fit into the cross-bridge theory. The relatively recent discovery of the giant elastic sarcomeric filament titin has thrust a previously missing element into any discussion of muscle function, in particular during active stretch. Indeed, the unexpected contribution of giant elastic proteins to muscle contractile function is highlighted by recent discoveries that twitchin-actin interactions are responsible for the "catch" property of invertebrate muscle. In this review, we examine several current theories that have been proposed to account for the properties of muscle during eccentric contraction. We ask how well each of these explains existing data and how an elastic filament can be incorporated into the sliding filament model. Finally, we review the increasing body of evidence for the benefits of including eccentric contractions into a program of muscle rehabilitation and strengthening.

Chronic eccentric arm cycling improves maximum upper-body strength and power.

INTRODUCTION: Eccentric leg cycling (cycle ergometry adapted to impose muscle lengthening contractions) offers an effective exercise for restoring lower-body muscular function, maintaining health, and improving performance in clinical and athletic populations. PURPOSE: We extended this model to the upper body and evaluated the effectiveness of a 7-week eccentric arm cycling (ECCarm) intervention to improve upper-body muscular function. We also explored whether ECCarm would alter arterial function. METHODS: Participants performed ECCarm (n = 9) or concentric arm cycling (CONarm; n = 8) 3×/week while training intensity increased (5-20 min, 60-70% upper-body peak heart rate). Maximum elbow extensor strength, upper-body concentric power, and peripheral and central arterial stiffness were assessed before and after training. RESULTS: During training, heart rates and perceived exertion did not differ between groups (~68% upper-body peak heart rate, ~12 Borg units, both P > 0.05), whereas power during ECCarm was ~2× that for CONarm (122 ± 43 vs. 59 ± 20 W, P < 0.01). Muscle soreness for ECCarm was greater than CONarm (P = 0.02), however, soreness was minimal for both groups (<0.50 cm). Following training, ECCarm exhibited greater changes in elbow extensor strength (16 ± 10 vs. 1 ± 9%, P = 0.01) and upper-body power (6 ± 8 vs. -3 ± 7%, P < 0.01) compared to CONarm. Peripheral and central arterial stiffness did not change for either group (both P > 0.05). CONCLUSION: Upper-body eccentric exercise improved dynamic muscular function while training at low exertion levels. Results occurred with minimal soreness and without compromising arterial function. ECCarm findings parallel eccentric leg cycling findings and indicate that eccentric cycle ergometry offers a robust model for enhancing upper-body muscular function. ECCarm could have applications in rehabilitation and sport training.

Physiological Mechanisms of Eccentric Contraction and Its Applications: A Role for the Giant Titin Protein.

When active muscles are stretched, our understanding of muscle function is stretched as well. Our understanding of the molecular mechanisms of concentric contraction has advanced considerably since the advent of the sliding filament theory, whereas mechanisms for increased force production during eccentric contraction are only now becoming clearer. Eccentric contractions play an important role in everyday human movements, including mobility, stability, and muscle strength. Shortly after the sliding filament theory of muscle contraction was introduced, there was a reluctant recognition that muscle behaved as if it contained an "elastic" filament. Jean Hanson and Hugh Huxley referred to this structure as the "S-filament," though their concept gained little traction. This additional filament, the giant titin protein, was identified several decades later, and its roles in muscle contraction are still being discovered. Recent research has demonstrated that, like activation of thin filaments by calcium, titin is also activated in muscle sarcomeres by mechanisms only now being elucidated. The mdm mutation in mice appears to prevent activation of titin, and is a promising model system for investigating mechanisms of titin activation. Titin stiffness appears to increase with muscle force production, providing a mechanism that explains two fundamental properties of eccentric contractions: their high force and low energetic cost. The high force and low energy cost of eccentric contractions makes them particularly well suited for athletic training and rehabilitation. Eccentric exercise is commonly prescribed for treatment of a variety of conditions including sarcopenia, osteoporosis, and tendinosis. Use of eccentric exercise in rehabilitation and athletic training has exploded to include treatment for the elderly, as well as muscle and bone density maintenance for astronauts during long-term space travel. For exercise intolerance and many types of sports injuries, experimental evidence suggests that interventions involving eccentric exercise are demonstrably superior to conventional concentric interventions. Future work promises to advance our understanding of the molecular mechanisms that confer high force and low energy cost to eccentric contraction, as well as signaling mechanisms responsible for the beneficial effects of eccentric exercise in athletic training and rehabilitation.

Huxleys' Missing Filament: Form and Function of Titin in Vertebrate Striated Muscle.

Although superthin filaments were inferred from early experiments on muscle, decades passed before their existence was accepted. Phylogenetic analyses suggest that titin, the largest known protein, first appeared in the common ancestor of chordates and nematodes and evolved rapidly via duplication. Twitchin and projectin evolved later by truncation. Sallimus mutants in Drosophila exhibit disrupted sarcomere and chromosome structure, suggesting that giant proteins may have evolved as chromosomal scaffolds that were co-opted for a similar purpose in striated muscles. Though encoded by only one gene, titin comprises hundreds of exons and has the potential for enormous diversity. Shorter isoforms typically confer greater passive stiffness associated with smaller in vivo muscle strains. Recent studies demonstrate unequivocally that titin stiffness increases upon muscle activation, but the mechanisms are only now being uncovered. Although some basic principles have been established, a vast opportunity remains to extend our understanding of titin function in striated muscle.

Skeletal muscle tissue in movement and health: positives and negatives.

The history of muscle physiology is a wonderful lesson in 'the scientific method'; our functional hypotheses have been limited by our ability to decipher (observe) muscle structure. The simplistic understanding of how muscles work made a large leap with the remarkable insights of A. V. Hill, who related muscle force and power to shortening velocity and energy use. However, Hill's perspective was largely limited to isometric and isotonic contractions founded on isolated muscle properties that do not always reflect how muscles function in vivo. Robert Josephson incorporated lengthening contractions into a work loop analysis that shifted the focus to dynamic muscle function, varying force, length and work done both by and on muscle during a single muscle work cycle. It became apparent that muscle is both a force generator and a spring. Titin, the missing filament in the sliding filament model, is a muscle spring, which functions very differently in cardiac versus skeletal muscle; its possible role in these two muscle types is discussed relative to their contrasting function. The good news for those of us who choose to work on skeletal muscle is that muscle has been reluctant to reveal all of its secrets.

Muscle force, work and cost: a novel technique to revisit the Fenn effect.

Muscle produces force by forming cross-bridges, using energy released from ATP. While the magnitude and duration of force production primarily determine the energy requirement, nearly a century ago Fenn observed that muscle shortening or lengthening influenced energetic cost of contraction. When work is done by the muscle, the energy cost is increased and when work is done on the muscle the energy cost is reduced. However, the magnitude of the 'Fenn effect' and its mirror ('negative Fenn effect') have not been quantitatively resolved. We describe a new technique coupling magnetic resonance spectroscopy with an in vivo force clamp that can directly quantify the Fenn effect [E=I+W, energy liberated (E) equals the energy cost of isometric force production (I) plus the work done (W)] and the negative Fenn effect (E=I-W) for one muscle, the first dorsal interosseous (FDI). ATP cost was measured during a series of contractions, each of which occurred at a constant force and for a constant duration, thus constant force-time integral (FTI). In all subjects, as the FTI increased with load, there was a proportional linear increase in energy cost. In addition, the cost of producing force greatly increased when the muscle shortened, and was slightly reduced during lengthening contraction. These results, though limited to a single muscle, contraction velocity and muscle length change, do quantitatively support the Fenn effect. We speculate that they also suggest that an elastic element within the FDI muscle functions to preserve the force generated within the cross-bridges.

Shiver me titin! Elucidating titin's role in shivering thermogenesis.

Shivering frequency scales predictably with body mass and is 10 times higher in a mouse than a moose. The link between shivering frequency and body mass may lie in the tuning of muscle elastic properties. Titin functions as a muscle 'spring', so shivering frequency may be linked to titin's structure. The muscular dystrophy with myositis (mdm) mouse is characterized by a deletion in titin's N2A region. Mice that are homozygous for the mdm mutation have a lower body mass, stiffer gait and reduced lifespan compared with their wild-type and heterozygous siblings. We characterized thermoregulation in these mice by measuring metabolic rate and tremor frequency during shivering. Mutants were heterothermic at ambient temperatures of 20-37°C while wild-type and heterozygous mice were homeothermic. Metabolic rate increased at smaller temperature differentials (i.e. the difference between body and ambient temperatures) in mutants than in non-mutants. The difference between observed tremor frequencies and shivering frequencies predicted by body mass was significantly larger for mutant mice than for wild-type or heterozygous mice, even after accounting for differences in body temperature. Together, the heterothermy in mutants, the increase in metabolic rate at low temperature differentials and the decreased tremor frequency demonstrate the thermoregulatory challenges faced by mice with the mdm mutation. Oscillatory frequency is proportional to the square root of stiffness, and we observed that mutants had lower active muscle stiffness in vitro. The lower tremor frequencies in mutants are consistent with reduced active muscle stiffness and suggest that titin affects the tuning of shivering frequency.

Eccentric exercise in rehabilitation: safety, feasibility, and application.

This nonexhaustive mini-review reports on the application of eccentric exercise in various rehabilitation populations. The two defining properties of eccentric muscle contractions--a potential for high muscle-force production at an energy cost that is uniquely low--are revisited and formatted as exercise countermeasures to muscle atrophy, weakness, and deficits in physical function. Following a dual-phase implementation, eccentric exercise that induces rehabilitation benefits without muscle damage, thereby making it both safe and feasible in rehabilitation, is described. Clinical considerations, algorithms of exercise progression, and suggested modes of eccentric exercise are presented.

Animal galloping and human hopping: an energetics and biomechanics laboratory exercise.

This laboratory exercise demonstrates fundamental principles of mammalian locomotion. It provides opportunities to interrogate aspects of locomotion from biomechanics to energetics to body size scaling. It has the added benefit of having results with robust signal to noise so that students will have success even if not "meticulous" in attention to detail. First, using respirometry, students measure the energetic cost of hopping at a "preferred" hop frequency. This is followed by hopping at an imposed frequency half of the preferred. By measuring the O2 uptake and work done with each hop, students calculate mechanical efficiency. Lessons learned from this laboratory include 1) that the metabolic cost per hop at half of the preferred frequency is nearly double the cost at the preferred frequency; 2) that when a person is forced to hop at half of their preferred frequency, the mechanical efficiency is nearly that predicted for muscle but is much higher at the preferred frequency; 3) that the preferred hop frequency is strongly body size dependent; and 4) that the hop frequency of a human is nearly identical to the galloping frequency predicted for a quadruped of our size. Together, these exercises demonstrate that humans store and recover elastic recoil potential energy when hopping but that energetic savings are highly frequency dependent. This stride frequency is dependent on body size such that frequency is likely chosen to maximize this function. Finally, by requiring students to make quantitative solutions using appropriate units and dimensions of the physical variables, these exercises sharpen analytic and quantitative skills.

How animals move: comparative lessons on animal locomotion.

Comparative physiology often provides unique insights in animal structure and function. It is specifically through this lens that we discuss the fundamental properties of skeletal muscle and animal locomotion, incorporating variation in body size and evolved difference among species. For example, muscle frequencies in vivo are highly constrained by body size, which apparently tunes muscle use to maximize recovery of elastic recoil potential energy. Secondary to this constraint, there is an expected linking of skeletal muscle structural and functional properties. Muscle is relatively simple structurally, but by changing proportions of the few muscle components, a diverse range of functional outputs is possible. Thus, there is a consistent and predictable relation between muscle function and myocyte composition that illuminates animal locomotion. When animals move, the mechanical properties of muscle diverge from the static textbook force-velocity relations described by A. V. Hill, as recovery of elastic potential energy together with force and power enhancement with activation during stretch combine to modulate performance. These relations are best understood through the tool of work loops. Also, when animals move, locomotion is often conveniently categorized energetically. Burst locomotion is typified by high-power outputs and short durations while sustained, cyclic, locomotion engages a smaller fraction of the muscle tissue, yielding lower force and power. However, closer examination reveals that rather than a dichotomy, energetics of locomotion is a continuum. There is a remarkably predictable relationship between duration of activity and peak sustainable performance.

Is titin a 'winding filament'? A new twist on muscle contraction.

Recent studies have demonstrated a role for the elastic protein titin in active muscle, but the mechanisms by which titin plays this role remain to be elucidated. In active muscle, Ca(2+)-binding has been shown to increase titin stiffness, but the observed increase is too small to explain the increased stiffness of parallel elastic elements upon muscle activation. We propose a 'winding filament' mechanism for titin's role in active muscle. First, we hypothesize that Ca(2+)-dependent binding of titin's N2A region to thin filaments increases titin stiffness by preventing low-force straightening of proximal immunoglobulin domains that occurs during passive stretch. This mechanism explains the difference in length dependence of force between skeletal myofibrils and cardiac myocytes. Second, we hypothesize that cross-bridges serve not only as motors that pull thin filaments towards the M-line, but also as rotors that wind titin on the thin filaments, storing elastic potential energy in PEVK during force development and active stretch. Energy stored during force development can be recovered during active shortening. The winding filament hypothesis accounts for force enhancement during stretch and force depression during shortening, and provides testable predictions that will encourage new directions for research on mechanisms of muscle contraction.

What is the role of titin in active muscle?

Several properties of muscle defy explanation solely based on the sliding filament-swinging cross-bridge theory. Indeed, muscle behaves as though there is a dynamic "spring" within the sarcomeres. We propose a new "winding filament" mechanism for how titin acts, in conjunction with the cross-bridges, as a force-dependent spring. The addition of titin into active sarcomeres resolves many puzzling muscle characteristics.

Muscle damage and muscle remodeling: no pain, no gain?

Skeletal muscle is a dynamic tissue that responds adaptively to both the nature and intensity of muscle use. This phenotypic plasticity ensures that muscle structure is linked to patterns of muscle use throughout the lifetime of an animal. The cascade of events that result in muscle restructuring - for example, in response to resistance exercise training - is often thought to be initiated by muscle damage. We designed this study to test the hypothesis that symptomatic (i.e. detectable) damage is a necessary precursor for muscle remodeling. Subjects were divided into two experimental populations: pre-trained (PT) and naive (NA). Demonstrable muscle damage was avoided in the PT group by a three-week gradual 'ramp-up' protocol. By contrast, the NA group was subjected to an initial damaging bout of exercise. Both groups participated in an eight-week high-force eccentric-cycle ergometry program (20 min, three times per week) designed to equate the total work done during training between the groups. The NA group experienced signs of damage, absent in the PT group, as indicated by greater than five times higher levels of plasma creatine kinase (CK) and self-reporting of initial perceived soreness and exertion, yet muscle size and strength gains were not different for the two groups. RT-PCR analysis revealed similar increases in levels of the growth factor IGF-1Ea mRNA in both groups. Likewise, the significant (P<0.01) increases in mean cross-sectional area (and total muscle volume) were equal in both groups. Finally, strength increases were identical for both groups (PT=25% and NA=26% improvement). The results of this study suggest that muscle rebuilding - for example, hypertrophy - can be initiated independent of any discernible damage to the muscle.

Electromyographic adaptations elicited by submaximal exercise in those naive to and in those adapted to eccentric exercise: a descriptive report.

The purpose of this study was to describe an electromyogram (EMG) pattern during a submaximal eccentric task in 7 subjects adapted to high-force chronic eccentric exercise and 6 subjects naive to eccentric exercise. The EMG in all subjects was quantified during identical submaximal (200 W) eccentric and concentric cycle ergometry tasks. The EMG of the eccentrically adapted subjects was decreased (p < 0.05) compared to the eccentrically naive subjects, in duration, amplitude, and intensity as evidenced by a decreased EMG during the pedal cycle. This decrease may be one component of the protective effect that results from progressively increasing repeated bouts of eccentric muscle work. Clients and patients transitioning to rigorous overload training should become adapted to high eccentric loads and forces to avoid injury and a potential delay in their strength and conditioning training regimens.