Muscle anatomy is a primary determinant of muscle relaxation dynamics in the lobster (<Emphasis Type="Italic">Panulirus interruptus</Emphasis>) stomatogastric system

We stained sarcomere thin filaments with fluorescently labeled phalloidin, measured sarcomere and muscle length, and calculated sarcomere number in pyloric and gastric mill muscles. A wide range of sarcomere lengths (3.25–12.29 μm), muscle lengths (5.9–21.1 mm), and sarcomere numbers (648–3,036) were observed. Sarcomere number differences occurred both because of changes in sarcomere length and muscle length, and sarcomere and muscle length varied independently. This independence, the wide range of sarcomere numbers present, and the muscles being all ‘slow’, graded muscles allowed us to use these data to test Huxley and Neidergerke’s (1954) hypothesis that muscle dynamics depend on sarcomere number. The time constants of exponential fits to contraction relaxations were used to measure muscle dynamics, and comparison of theoretical predictions and experimental results quantitatively confirm the predicted dependence. The differing dynamics of the various pyloric muscles are likely functionally important, and the dependence of muscle dynamics on sarcomere number implies that sarcomere number is likely closely regulated in these muscles. The stomatogastric system may thus be an excellent model system for studying the mechanisms regulating muscle sarcomere number.     

Measuring muscle and joint geometry parameters of a shoulder for modeling purposes.

An extensive set of muscle and joint geometry parameters was measured of the right shoulder of an embalmed male. For all muscles the optimal muscle fiber length was determined by laser diffraction measurements of sarcomere length. In addition, tendon length and physiological cross-sectional area were determined. The parameter set was needed to enhance the reliability of a computer model of the shoulder (Van der Helm, 1994a,b Journal of Biomechanics 27, 527-550, 551-569). With the model, an abduction of the arm was simulated in seven positions, at 30 degrees intervals. In each of the simulated arm positions, actual sarcomere lengths were calculated from the lengths of 104 muscle elements, distributed over 16 shoulder muscles. For most muscle elements, the simulated abduction appeared to take place within the sarcomere length range in which the muscle elements can exert force. The muscle elements can then act on the ascending limb as well as on the plateau and on the descending limb of the relative force-length curves of sarcomeres. The produced data set is not only important for the refinement of shoulder modeling, but also for functional analyses of shoulder movements in general.     

Sarcomere length organization as a design for cooperative function amongst all lumbar spine muscles

The functional design of spine muscles in part dictates their role in moving, loading, and stabilizing the lumbar spine. There have been numerous studies that have examined the isolated properties of these individual muscles. Understanding how these muscles interact and work together, necessary for the prediction of muscle function, spine loading, and stability, is lacking. The objective of this study was to measure sarcomere lengths of lumbar muscles in a neutral cadaveric position and predict the sarcomere operating ranges of these muscles throughout full ranges of spine movements. Sarcomere lengths of seven lumbar muscles in each of seven cadaveric donors were measured using laser diffraction. Using published anatomical coordinate data, superior muscle attachment sites were rotated about each intervertebral joint and the total change in muscle length was used to predict sarcomere length operating ranges. The extensor muscles had short sarcomere lengths in a neutral spine posture and there were no statistically significant differences between extensor muscles. The quadratus lumborum was the only muscle with sarcomere lengths that were optimal for force production in a neutral spine position, and the psoas muscles had the longest lengths in this position. During modeled flexion the extensor, quadratus lumborum, and intertransversarii muscles lengthened so that all muscles operated in the approximate same location on the descending limb of the force-length relationship. The intrinsic properties of lumbar muscles are designed to complement each other. The extensor muscles are all designed to produce maximum force in a mid-flexed posture, and all muscles are designed to operate at similar locations of the force-length relationship at full spine flexion.     

Energetics of activation in frog skeletal muscle at sarcomere lengths beyond myofilament overlap.

Experiments were designed to gain information about the effects of extremely long sarcomere lengths (greater than 3.8 microns) on muscle activation. The amount of energy liberated in an isometric twitch by muscles stretched to sarcomere lengths where myofilament overlap is vanishingly small (greater than 3.6 microns) is thought to be an indirect measure of the Ca2+ cycled during contraction. The effects of altering sarcomere length from 3.8 to 4.3 microns on the amount of Ca2+ cycled was measured using twitch energy liberation as an indicator of the Ca2+ cycled. Twitch energy liberation decreased by approximately 20% over this sarcomere length region, suggesting that the amount of Ca2+ released by a single action potential is not altered dramatically when a muscle is stretched to extreme lengths.     

The relation between Z--disk lattice spacing and sarcomere length in sartorius muscle fibres from Hyla cerulea.

Sartorius muscles from the green tree frog Hyla cerulea were set at variety of muscle lengths and fixed for electron microscopy using acrolein followed by osmium tetroxide. The sarcomere length, s, was determined in thick sections using laser-diffraction. The Z-disk lattice spacing, z, was measured in electron micrographs of thin sections from the same muscles. The Z-disk lattice was found to expand as sarcomere length decreased such that the quantity sz2 was constant at 1-05 X 10(6) nm3 for all sarcomere lengths in the range 1-9-2-9 mum. Thus, the sarcomere length dependency of the Z-disk lattice is similar to that of the myosin filament lattice. The density of thin filaments per unit cross section of fibril leaving the Z-disk is less than their density in the A band. Thus, fibrils have a smaller cross section in the I band, leaving more inter-fibrillar space there. This may explain why more mitochondria and lipid droplets are located in the I bands than in the A bands. It is suggested that the Z-disk may contributed to the short range elasticity of muscle fibres.     

A novel muscle biopsy clamp yields accurate in vivo sarcomere length values

The measurement of in vivo muscle sarcomere length facilitates the definition of in vivo muscle functional properties and comparison of muscle designs amongst functional muscle groups. In vivo sarcomere lengths are available for just a handful of human muscles, largely due to the technical challenges associated with their measurement. The purpose of this report was to develop and test a muscle biopsy clamp that can quickly and accurately measure in vivo muscle sarcomere length. To test the device, muscle biopsies (n=23) were removed from the tibialis anterior muscles of New Zealand White rabbits immediately after sarcomere length measurements were made using laser diffraction. The muscle biopsy contained within the clamp was immediately fixed in Formalin for subsequent sarcomere length measurement. Comparisons of clamp-based and diffraction-based sarcomere lengths demonstrated excellent agreement between the two techniques, especially when the biopsy was obtained at relatively long lengths (above 2.6 μm). Given the intraoperative speed and simplicity of this technique and the relatively low-cost of the biopsy clamp, this method of measuring muscle sarcomere length should help investigators generate much-needed in vivo muscle structural and functional data.     

Polarized light observations on striated muscle contraction in a mite

Contraction of individual sarcomeres within the living mite Tarsonemus sp. was observed by polarized light microscopy. In unflattened animals the usual range of contraction was such that the minimum sarcomere length approximated the length of the A region, and the maximum sarcomere length was about twice the length of the A region. The central sarcomeres of the dorsal metapodosomal muscles were observed in detail. The A band length increased slightly with increasing sarcomere length since the regression of I region length on sarcomere length had an average slope of 0.91. When the A band length in a sarcomere which was shortening was compared with the length when the same sarcomere lengthened, no significant difference was seen. The A band of each sarcomere seemed to act as a not too rigid limit to further shortening; this agreed with the reversible shortening of a muscle in which the A band had been experimentally shortened. An H region was visible at long sarcomere lengths and was not visible at short sarcomere lengths, even when the muscle was actively shortening. The rate of change of H region length with sarcomere length suggested that I filament length may increase as sarcomere length increases. Despite this effect and the small increase in A length with sarcomere length, the results are considered to be consistent with a model in which shortening occurs by the relative movement of A and I filaments, with little or no change in length of either set of filaments. Sarcomere shortening was clearly associated with an increase in the retardation of the A region.     

Resonant Reflection Spectroscopy of Biomolecular Arrays in Muscle

Sarcomeres, the functional units of contraction in striated muscle, are composed of an array of interdigitating protein filaments. Direct interaction between overlapping filaments generates muscular force, which produces animal movement. When filament length is known, sarcomere length successfully predicts potential force, even in whole muscles that contain billions of sarcomere units. Inability to perform in vivo sarcomere measurements with submicrometer resolution is a long-standing challenge in the muscle physiology field and has hampered studies of normal muscle function, adaptation, injury, aging, and disease, particularly in humans. Here, we develop theory and demonstrate the feasibility of to our knowledge a new technique that measures sarcomere length with submicrometer resolution. In this believed novel approach, we examine sarcomere structure by measuring the multiple resonant reflections that are uniquely defined by Fourier decomposition of the sarcomere protein spatial framework. Using a new supercontinuum spectroscopic system, we show close agreement between sarcomere lengths measured by resonant reflection spectroscopy and laser diffraction in an ensemble of 10 distinct muscles.     

Reconstruction of the human gastrocnemius force-length curve in vivo: part 1-model-based validation of method

The muscle fiber force-length relationship has been explained in terms of the cross-bridge theory at the sarcomere level. In vivo, for a physiologically realistic range of joint motion, and therefore range of muscle fiber lengths, only part of the force-length curve may be used; that is, the section of the force-length curve expressed can vary. The purpose of this study was to assess the accuracy of a method for determining the expressed section of the force-length curve for biarticular muscles. A muscle model was used to simulate the triceps surae muscle group. Three model formulations were used so that the gastrocnemius operated over different portions of the force-length curve: the ascending limb, the plateau region, and the descending limb. Joint moment data were generated for a range of joint configurations and from this simulated data the region of the force- length relationship that the gastrocnemius muscle operated over was successfully reconstructed using the algorithm of Herzog and ter Keurs (1988a). Further simulations showed that the correct region of the force-length curve was accurately reconstructed even in the presence of random and systematic noise generated to reflect the effects of sampling errors, and incomplete muscle activation.     

Non-specific termination of simian virus 40 DNA replication.

Axial X-ray diffraction patterns have been studied from relaxed, contracted and rigor vertebrate striated muscles at different sarcomere lengths to determine which features of the patterns depend on the interaction of actin and myosin. The intensity of the myosin layer lines in a live, relaxed muscle is sometimes less in a stretched muscle than in the muscle at rest-length; the intensity depends not only on the sarcomere length but on the time that has elapsed since dissection of the muscle. The movement of cross-bridges giving rise to these intensity changes are not caused solely by the withdrawal of actin from the A-band.When a muscle contracts or passes into rigor many changes occur that are independent of the sarcomere length: the myosin layer lines decrease in intensity to about 30% of their initial value when the muscle contracts, and disappear completely when the muscle passes into rigor. Both in contracting and rigor muscles at all sarcomere lengths the spacings of the meridional reflections at 143 Å and 72 Å are 1% greater than from a live relaxed muscle at rest-length. It is deduced that the initial movement of cross-bridges from their positions in resting muscle does not depend on the interaction of each cross-bridge with actin, but on a conformational change in the backbone of the myosin filament: occurring as a result of activation. The possibility is discussed that the conformational change occurs because the myosin filament, like the actin filament, has an activation control mechanism. Finally, all the X-ray diffraction patterns are interpreted on a model in which the myosin filament can exist in one of two possible states: a relaxed state which gives a diffraction pattern with strong myosin layer lines and an axial spacing of 143.4 Å, and an activated state which gives no layer lines but a meridional spacing of 144.8 Å.     

Active force-length relationship of human lower-leg muscles estimated from morphological data: a comparison of geometric muscle models

Muscle fibre lengths, pennation angles, and sarcomere lengths were measured (the latter by a diffraction technique) for each of the muscles of three embalmed lower-leg specimens. From these data and filament lengths from Walker & Schrodt (1973), the optimum fibre lengths were determined. Relationships between length and active force (at full activation) of the lower-leg muscles were calculated by use of (i) a unipennate muscle model, (ii) a bipennate model, and (iii) bipennate models in which the cosine of the pennation angle is approximated as length independent. It is concluded that the first two models are equally useful and that the use of the last models is discouraged in case of strongly pennated muscles. Non-uniformity of fibre parameters within one muscle appears to have little effect on the force-length relationship.     

FILAMENT LENGTHS IN STRIATED MUSCLE

Filament lengths in resting and excited frog muscles have been measured in the electron microscope, and investigations made of the changes in length that are found under different conditions, to distinguish between those changes which arise during preparation and the actual differences in the living muscles. It is concluded that all the measured differences in filament length are caused by the preparative procedures in ways that can be simply accounted for, and that the filament lengths are the same in both resting and excited muscles at all sarcomere lengths greater than 2.1 µ, viz., A filaments, 1.6 µ; I filaments, 2.05 µ. The fine periodicity visible along the I filaments also has been measured in frog, toad, and rabbit muscles and found to be 406 A.     

Myofilament lengths of cat skeletal muscle: theoretical considerations and functional implications.

The cat is the primary model for neuromuscular research. However, sarcomere geometry, in particular thin-myofilament lengths of cat skeletal muscles, is not known, thus preventing adequate muscle modeling on the sarcomere level. The purpose of this study was to determine thin-myofilament lengths in cat skeletal muscle. It was found that average thin-myofilament lengths of cat tibialis anterior muscles (1.12 microns) were larger than the average values reported for frog (approximately 0.95 microns), rat (1.09 microns), and rabbit muscles (1.09 microns) and were smaller than the values reported for monkey (1.16 microns) and human skeletal muscles (1.27 microns). According to the cross-bridge theory of muscular contraction, this result implies that the range of sarcomere length on the ascending limb of the force-length relation for cat muscle is between those of frog, rat, and rabbit on the one side and monkey and human on the other side. It is speculated that the differences in thin-myofilament lengths of different animals are related to the functional demands of these muscles in everyday movement tasks. Isolated experimental observations appear to support this speculation.     

Sarcomere shortening in pressure overload hypertrophy

Sarcomere shortening during contraction was measured by using laser diffraction, in thin, rabbit right ventricular (RV) trabeculae from normal hearts (N) (n = 5) and from hearts subjected to RV pressure overload by pulmonary banding (H) (n = 5). Banding resulted in substantial RV hypertrophy after 2 wk. Hypertrophied preparations had the same resting muscle length (H = 3.15 +/- 0.29 mm) and resting sarcomere lengths (H = 2.16 +/- 0.005 micron) as the normal preparations (3.10 +/- 0.37 mm, 2.16 +/- 0.008 micron, respectively). Total tension at the peak of isometric twitches was the same as normal in the hypertrophied muscles (N = 8.06 +/- 1.20, H = 8.51 +/- 1.95 g/mm2). However, the amount of auxotonic sarcomere shortening was much less than normal in the hypertrophied preparations (N = 0.39 +/- 0.028, H = 0.19 +/- 0.034 micron; P less than 0.001). In isotonic contractions in which the ratio of muscle shortening to resting muscle length was the same in both the normal and hypertrophied muscles (ratio of 0.05 in both groups), the extent of sarcomere shortening relative to resting sarcomere length was less in the hypertrophied muscles than in the normal preparations (N = 0.14 +/- 0.01), H = 0.07 +/- 0.01; P less than 0.01). Series elasticity was the same as normal in the hypertrophied muscle P less than 0.05). Less auxotonic sarcomere shortening for a given level of isometric tension development and less isotonic sarcomere shortening per unit muscle shortening indicate that there is less than normal work per sarcomere during contraction in hypertrophied myocardium. These findings may have important implications for intracellular compensatory adaptation in pressure overload cardiac hypertrophy.     

The muscle line of action in current models of the human cervical spine: a comparison with in vivo MRI data

Cervical muscles are commonly represented by line-of-action models. This investigation evaluates the performance of three types of model implementations, based on their ability to mimic geometric in-vivo aspects of muscles. Five prominent pairs of neck muscles were reconstructed in three head positions using magnetic resonance imaging. Based on the reconstructions, muscle approximations were created that represent the muscles with piecewise straight lines. Measured and modelled muscle approximations were compared with respect to their pulling directions at the attachment sites and the overall distance between the muscle paths. Muscle lengths were evaluated in two ways. First, length discrepancies were determined between measured and modelled muscles depending on the head position. Second, the difference of muscle lengths in neutral and deflected head positions for measurement and models were calculated. The results indicate considerable differences between models and measurements. Pulling directions, for instance, differed by up to 40°, depending on the chosen muscle and the type of muscle implementation.     

Capillary tortuosity in skeletal muscles of mammals depends on muscle contraction

Capillary orientation (anisotropy) was compared in hindlimb muscles of mammals of different size and/or different aerobic capacity (dog, goat, pony, and calf). All muscles were fixed by vascular perfusion at sarcomere lengths ranging from 1.5 to 2.7 micron. The ratios of capillary counts per fiber cross-sectional area on two sets of sections (0 and 90 degrees) to the muscle fiber axis were used to estimate capillary anisotropy and the coefficient c(K,0) relating 1) capillary counts on transverse sections (a commonly used parameter to assess muscle capillarity) and 2) capillary length per volume of fiber (i.e., capillary length density). Capillary orientation parallel to the muscle fiber axis decreased substantially with muscle fiber shortening. In muscles fixed at sarcomere lengths of 2.69 microns (dog vastus intermedius) and 1.52 microns (dog gastrocnemius), capillary tortuosity and branching added 7 and 64%, respectively, to capillary length density. The data obtained in this study are highly consistent with the previously demonstrated relationship between capillary anisotropy and sarcomere length in extended vs. contracted rat muscles, by use of the same method. Capillary anisotropy in mammalian locomotory muscles is curvilinearly related to sarcomere length. No systematic difference was found in capillary tortuosity with either body size, athletic ability, or aerobic capacity. Capillary tortuosity is a consequence of fiber shortening rather than an indicator of the O2 requirements of the tissue.     

Interaction between series compliance and sarcomere kinetics determines internal sarcomere shortening during fixed-end contraction

The interaction between contractile force and in-series compliance was investigated for the intact skeletal muscle-tendon unit (MTU) of Rana pipiens semitendinosus muscles during fixed-end contraction. It was hypothesized that internal sarcomere shortening is a function of the length-force characteristics of contractile and series elastic components. The MTUs (n=18) were dissected, and, while submerged in Ringer's solution, muscles were activated at nine muscle lengths (-2 to +6 mm relative to optimal length in 1 mm intervals), while measuring muscle force and sarcomere length (SL) by laser diffraction. The MTU was clamped either at the bone (n=6), or at the proximal and distal ends of the aponeuroses (n=6). Muscle fibers were also trimmed along with aponeuroses down to 5-20 fibers and identical measurements were performed (n=6). The magnitude of shortening decreased as MTU length increased. The magnitude of shortening ranged from -0.08 to 0.3 microm, and there was no significant difference between delta SL as a function of clamp location. When aponeuroses were trimmed, sarcomere shortening was not observed at L(0) and longer. These results suggest that the aponeurosis is the major contributor to in-series compliance. Results also support our hypothesis but there also appear to be other factors affecting internal sarcomere shortening. The functional consequence of internal sarcomere shortening as a function of sarcomere length was to skew the muscle length-tension relationship to longer sarcomere lengths.     

Differential serial sarcomere number adaptations in knee extensor muscles of rats is contraction type dependent.

Sarcomerogenesis, or the addition of sarcomeres in series within a fiber, has a profound impact on the performance of a muscle by increasing its contractile velocity and power. Sarcomerogenesis may provide a beneficial adaptation to prevent injury when a muscle consistently works at long lengths, accounting for the repeated-bout effect. The association between eccentric exercise, sarcomerogenesis and the repeated-bout effect has been proposed to depend on damage, where regeneration allows sarcomeres to work at shorter lengths for a given muscle-tendon unit length. To gain additional insight into this phenomenon, we measured fiber dynamics directly in the vastus lateralis (VL) muscle of rats during uphill and downhill walking, and we measured serial sarcomere number in the VL and vastus intermedius (VI) after chronic training on either a decline or incline grade. We found that the knee extensor muscles of uphill walking rats undergo repeated active concentric contractions, and therefore they suffer no contraction-induced injury. Conversely, the knee extensor muscles during downhill walking undergo repeated active eccentric contractions. Serial sarcomere numbers change differently for the uphill and downhill exercise groups, and for the VL and VI muscles. Short muscle lengths for uphill concentric-biased contractions result in a loss of serial sarcomeres, and long muscle lengths for downhill eccentric-biased contractions result in a gain of serial sarcomeres.     

The muscle line of action in current models of the human cervical spine: a comparison with in vivo MRI data

Cervical muscles are commonly represented by line-of-action models. This investigation evaluates the performance of three types of model implementations, based on their ability to mimic geometric in-vivo aspects of muscles. Five prominent pairs of neck muscles were reconstructed in three head positions using magnetic resonance imaging. Based on the reconstructions, muscle approximations were created that represent the muscles with piecewise straight lines. Measured and modelled muscle approximations were compared with respect to their pulling directions at the attachment sites and the overall distance between the muscle paths. Muscle lengths were evaluated in two ways. First, length discrepancies were determined between measured and modelled muscles depending on the head position. Second, the difference of muscle lengths in neutral and deflected head positions for measurement and models were calculated. The results indicate considerable differences between models and measurements. Pulling directions, for instance, differed by up to 40°, depending on the chosen muscle and the type of muscle implementation.