Light diffraction studies of sarcomere dynamics in single skeletal muscle fibers.

A position-sensitive optical diffractometer has been used to examine the diffraction spectra produced by single skeletal muscle fibers during twitch and tetanic contraction. First-order diffraction lines were computer-analyzed for mean sarcomere length, line intensity, and percent dispersion in sarcomere length. Line intensity was observed to decrease rapidly by about 60 percent during a twitch, with an exponential recovery to resting intensity persisting well beyond cessation of sarcomere shortening; recovery was particularly prolonged at zero myofilament overlap. A number of single fibers at initial lengths from 2.5 to 3.5 MICRON EXHIBITED a splitting of the first-order line into two or more components during relaxation, with components merging back into a single peak by 200 ms after stimulation. This splitting reflects the asynchronous nature of myofibrillar relaxation within a single fiber. During tetanus, the dispersion decreased by more than 10 percent from onset to plateau, implying a gradual stabilization of sarcomeres.     

Theory of light diffraction by single skeletal muscle fibers.

A theoretical discussion is presented describing the diffraction of laser light by a single fiber of striated muscle. The complete three-dimensional geometry of the fiber has been taken into consideration. The basic repeated unit is taken as the sarcomere of a single myofibril, including its cylindrical geometry. The single fiber is considered as the sum of myofibrils up to the fiber dimensions. When proper phasing is taken into account, three cases of interest are analyzed. (a) When the adjacent myofibrils are totally aligned with respect to their index of refraction regions (e.g., A and I bands), then the diffraction pattern reflects that of a larger striated cylinder with the dimensions of the fiber. (b) When a particular skew plane develops for the myofibril elements, additional Bragg reflection occurs at certain specific sarcomere lengths, and intensity asymmetry amongst the diffracted orders occurs. (c) When the myofibril phasing changes in a random fashion, while all sarcomeres remain at the same length, then intensity decrease is directly related to the phase deviation from a reference phase point. This condition may well describe a fiber undergoing active isometric contraction.     

Fine structure of single fibres of human skeletal muscle

Summary Individual muscle fibres were separated from freeze-dried needle biopsies and classed as type I or type II fibres according to their myofibrillar ATP-ase. Portions of the same fibres were processed for electron microscopy and their fine structure examined. Type I fibres were found to have thicker Z-bands and more mitochondria and lipid droplets than the type II fibres.     

Modulation of passive force in single skeletal muscle fibres

In this study, we investigated the effects of activation and stretch on the passive force-sarcomere length relationship in skeletal muscle. Single fibres from the lumbrical muscle of frogs were placed at varying sarcomere lengths on the descending limb of the force-sarcomere length relationship, and tetanic contractions, active stretches and passive stretches (amplitudes of ca 10% of fibre length at a speed of 40% fibre length/s) were performed. The passive forces following stretch of an activated fibre were higher than the forces measured after isometric contractions or after stretches of a passive fibre at the corresponding sarcomere length. This effect was more pronounced at increased sarcomere lengths, and the passive force-sarcomere length relationship following active stretch was shifted upwards on the force axis compared with the corresponding relationship obtained following isometric contractions or passive stretches. These results provide strong evidence for an increase in passive force that is mediated by a length-dependent combination of stretch and activation, while activation or stretch alone does not produce this effect. Based on these results and recently published findings of the effects of Ca2+ on titin stiffness, we propose that the observed increase in passive force is caused by the molecular spring titin.     

X-ray diffraction studies of the contractile mechanism in single muscle fibres

The molecular mechanism of muscle contraction was investigated in intact muscle fibres by X-ray diffraction. Changes in the intensities of the axial X-ray reflections produced by imposing rapid changes in fibre length establish the average conformation of the myosin heads during active isometric contraction, and show that the heads tilt during the elastic response to a change in fibre length and during the elementary force generating process: the working stroke. X-ray interference between the two arrays of myosin heads in each filament allows the axial motions of the heads following a sudden drop in force from the isometric level to be measured in situ with unprecedented precision. At low load, the average working stroke is 12 nm, which is consistent with crystallographic studies. The working stroke is smaller and slower at a higher load. The compliance of the actin and myosin filaments was also determined from the change in the axial spacings of the X-ray reflections following a force step, and shown to be responsible for most of the sarcomere compliance. The mechanical properties of the sarcomere depend on both the motor actions of the myosin heads and the compliance of the myosin and actin filaments.     

Optical polarization properties of the diffraction spectra from single fibers of skeletal muscle.

The diffraction spectra of laser light from single fibers of skeletal muscle exhibit a large degree of optical depolarization. When the linearly polarized incident laser source is oriented at polarization angles between 0 less than theta less than pi/2 rad with respect to the fiber axis, the diffracted light is elliptically polarized. These results show that the phase angle of the ellipse rotates by as much as 20 degrees when the fiber is stretched from 2.4 to 3.8 microns. To further ascertain that the observed phenomenon is diffraction related, an experiment monitoring the spectra of scattered light in between diffraction orders showed this signal to be significantly more linearly polarized. These results suggest that the degree of elliptical polarization of the diffraction spectra is a sensitive probe of A-band dynamics, including changes of the anisotropic S-2 elements.     

Light diffraction studies of single muscle fibers as a function of fiber rotation

Light diffraction patterns from single glycerinated frog semitendinosus muscle fibers were examined photographically and photoelectrically as a function of diffraction angle and fiber rotation. The total intensity diffraction pattern indicates that the order maxima change both position and intensity periodically as a function of rotation angle. The total diffracted light, light diffracted above and below the zero-order plane, and light diffracted into individual orders gives information about the fiber's longitudinal and rotational structure and its noncylindrical symmetry.     

Eccentric and posteccentric contractile behaviour of skeletal muscle a comparative study in frog single fibres and in humans

Eccentric and posteccentric force behaviour in human skeletal muscle and in isolated frog muscle fibres was studied by imposing stretch-and-hold loading conditions during contractions with maximal voluntary effort or under tetanic stimulation in the isolated preparations. The investigations on human muscle were made on the forearm flexors of a group of kayak racers (n = 16; age: 17-22 years) and of schoolgirls (n = 15; age: 17-18 years) with both groups participating in a strength-training programme over 4 (kayak racers) or 3 (girls) months. Half of the training regime consisted of eccentric elements. In the isolated muscle fibres, it could be shown that in the posteccentric hold phase the enhanced force decayed exponentially to the original isometric value with a mean time-constant of 0.35 s (10 degrees C) and of 0.23 (20 degrees C). In the forearm flexor of human subjects similar results were obtained not only qualitatively but even quantitatively (time constant of posteccentric force decay: 0.25-0.37 s). Strength training in both groups did not lead to an enhancement in maximal isometric force alone [mean increase in force 17 (SD 10)%], a well-known and generally accepted fact, but also to a parallel shift in eccentric [21 (SD 10)%] and posteccentric force level. The close similarity between the findings in isolated muscle fibres and in human muscle in situ suggests that the eccentric and posteccentric behaviour must be primarily ascribed to the contractile properties of the muscle fibres themselves. A three-element muscle model with variable visco-elastic properties would appear to be most suitable for simulating the experimental findings.     

Analysis of equatorial x-ray diffraction patterns from skeletal muscle.

Some of the factors that affect the intensities and the phases of the first five equatorial x-ray reflections from skeletal muscle are studied by simplified models describing axially projected mass distributions in unit cells. Examples of mass distributions that produce various phase combinations and intensities are presented. Effects due to radial movement of crossbridges and those due to mass transfer between the thick filament and the thin filament regions are compared. In addition, the study suggests that some features in the reconstructed filament structures could be due to the consequences of limited resolution.     

Force-velocity relationship in single muscle fibres

In recent papers, it has been shown experimentally that the force-velocity relationship in single muscle fibres presents deviations from hyperbolicity at high values of the load (Edman, Mulieri & Scubon-Mulieri, 1976; Edman & Hwang,1977). It has been shown independently and on theoretical bases, that the parameter “b” in Hill's characteristic equation also presents deviations from its normal value at low values of the speed of shortening, i.e. at high values of the load (Morel, Pinset-Härström & Gingold, 1976). In the present paper, it is shown that both the experimental and the theoretical results are in excellent agreement and a theoretical force-velocity relationship is proposed.     

Decrease in light diffraction intensity of contracting muscle fibres

Single fibres from the semitendinosus muscle of frog were illuminated normally with a He–Ne laser. The intensity transient and fine structure pattern of light diffracted from the fibre undergoing isometric twitches were measured. During fibre shortening, the intensity decreased rapidly and the fine structure pattern preserved its shape and moved swiftly away from the undiffracted laser beam. The fine structure patterns of the contracting and resting fibre were nearly identical. The ratio of intensities of the contracting and resting fibre of the same sarcomere length was determined as a function of the time elapsed after fibre stimulation. The time-resolved intensity ratio increased with sarcomere length and became unity when sarcomere length was between 3.5 m and 3.7 m. A diffraction theory based on the sarcomere unit was developed. It contained a parameter describing the strength of filament interaction. The comparison between the theory and data shows that the initial intensity drop during contraction is primarily due to filament interactions. At a later stage of contraction, sarcomere disorder becomes the major component causing the intensity to decrease. Diffraction models which use the Debye-Waller formalism to explain the intensity decrease are discussed. The sarcomere-unit diffraction model is applied to previously reported intensity measurements from active fibres.     

Further histochemical properties of rabbit skeletal muscle fibres

Summary The histochemical activities of succinic dehydrogenase (SDH), creatine kinase (CK), sarcoplasmic reticular ATPase (SR-ATPase) and myosin ATPase were studied in serial sections of rabbit adductor muscle. Three fibre types were distinguished depending upon the distribution of the enzyme activities. The type II white fibres posessing minimal SDH showed high myosin ATPase, SR-ATPase and ATPase dependent CK activities. Red oxidative fibres showing high SDH fell into two distinct groups: One category had mainly a peripheral localization of SDH and showed an enzymatic profile identical to that of type II white fibres. The second category of red fibres displayed both a homogeneous distribution of small diformazan granules throughout the fibre as well as a sub-sarcolemmal collection when tested for SDH activity but possessed very low amounts of reaction product of the various enzymes of the energetic metabolism studied. Since it is well established that the myosin ATPase of a fibre correlates with its contraction time, the present histochemical investigation provides further support for this concept by demonstrating the presence of high SR-ATPase and ATPase dependent CK activities in all white and red fibres rich in myosin ATPase.     

Plasticity of skeletal muscle fibres in space-flown primates

Skeletal muscle atrophy and fibre type transitions were observed as a rule in rats exposed to micro- and zero-gravity, flown on boards of biosatellites and space shuttle ships. Much less is known about the spaceflight-induced muscle events in primates. The latter are animals of special interest since pattern of their onground motor activities works in ways alike to the human one, though the opportunities of studies are much wider. One of the targets of the study was to investigate the influence of spaceflight conditions on tissue morphology in monkey skeletal muscles of different functional and structural organization.