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.     

Chronic coexistence of two troponin T isoforms in adult transgenic mouse cardiomyocytes decreased contractile kinetics and caused dilatative remodeling

Our previous in vivo and ex vivo studies suggested that coexistence of two or more troponin T (TnT) isoforms in adult cardiac muscle decreased cardiac function and efficiency (Huang QQ, Feng HZ, Liu J, Du J, Stull LB, Moravec CS, Huang X, Jin JP, Am J Physiol Cell Physiol 294: C213-C22, 2008; Feng HZ, Jin JP, Am J Physiol Heart Circ Physiol 299: H97-H105, 2010). Here we characterized Ca(2+)-regulated contractility of isolated adult cardiomyocytes from transgenic mice coexpressing a fast skeletal muscle TnT together with the endogenous cardiac TnT. Without the influence of extracellular matrix, coexistence of the two TnT isoforms resulted in lower shortening amplitude, slower shortening and relengthening velocities, and longer relengthening time. The level of resting cytosolic Ca(2+) was unchanged, but the peak Ca(2+) transient was lowered and the durations of Ca(2+) rising and decaying were longer in the transgenic mouse cardiomyocytes vs. the wild-type controls. Isoproterenol treatment diminished the differences in shortening amplitude and shortening and relengthening velocities, whereas the prolonged durations of relengthening and Ca(2+) transient in the transgenic cardiomyocytes remained. At rigor state, a result from depletion of Ca(2+), resting sarcomere length of the transgenic cardiomyocytes became shorter than that in wild-type cells. Inhibition of myosin motor diminished this effect of TnT function on cross bridges. The length but not width of transgenic cardiomyocytes was significantly increased compared with the wild-type controls, corresponding to longitudinal addition of sarcomeres and dilatative remodeling at the cellular level. These dominantly negative effects of normal fast TnT demonstrated that chronic coexistence of functionally distinct variants of TnT in adult cardiomyocytes reduces contractile performance with pathological consequences.     

Measurement of sarcomere shortening in skinned fibers from frog muscle by white light diffraction.

A new optical-electronic method has been developed to detect striation spacing of single muscle fibers. The technique avoids Bragg-angle and interference-fringe effects associated with laser light diffraction by using polychromatic (white) light. The light is diffracted once by an acousto-optical device and then diffracted again by the muscle fiber. The double diffraction reverses the chromatic dispersion normally obtained with polychromatic light. In frog skinned muscle fibers, active and passive sarcomere shortening were smooth when observed by white light diffraction, whereas steps and pauses occurred in the striation spacing signals obtained with laser illumination. During active contractions skinned fibers shortened at high rates (3-5 microns/s per half sarcomere, 0-5 degrees C) at loads below 5% of isometric tension. Compression of the myofibrillar lateral filament spacing using osmotic agents reduced the shortening velocity at low loads. A hypothesis is presented that high shortening velocities are observed with skinned muscle fibers because the cross-bridges cannot support compressive loads when the filament lattice is swollen.     

High resolution measurement of striation patterns and sarcomere motions in cardiac muscle cells.

We describe an extension of the method of Myers et al. (1982) to measure with high precision the uniformity of contractile motions that occur between sarcomeres in the isolated cardiac muscle cell (guinea pig and rat). The image of the striations, observed with modulation contrast microscopy, was detected by a linear array of photodiodes. Sarcomere length was measured greater than 500/s from the frequency of the array's video signal at two selectable regions of the cell. A precision test grating demonstrated that method resolves known differences in the spacing between two contiguous striations to +/- 0.01 micron and that the effects of image translation and microscopic resolution are minor. The distribution of striation spacing appears to be discrete in isolated segments of the cell, and patches of fairly uniform length can be identified that are laterally contiguous. When electrically triggered, contraction is synchronous and the sarcomeres shorten and relengthen smoothly. The contrast between the striations is transiently enhanced during relengthening, an indication that the contracting cell can not be treated as a simple grating. Pauses that occur during late in relengthening (and transient contractile alternans) are characterized by very synchronized activity. These forms of irregular contractile behavior are not explained by desynchronization of a mechanism of release of intracellular calcium. A companion article describes application of the technique to study the nonuniform motions that occur between sarcomeres.     

Efficiency of light diffraction by cross-striated muscle fibers under stretch and during isometric contraction.

When light is diffracted by a single frog muscle fiber the intensities I kappa of the different orders kappa (kappa = 1,2,3) strongly depend on the angle between the axis of the incident beam and the fiber axis. Maximum intensity is not obtained with perpendicular incidence (omega = 0 degree) but at angles that can be calculated for each order number and sarcomere length using Bragg's formula. In analogy to techniques developed for x-ray structure analysis of mosaic crystals we have rotated the fiber around an axis perpendicular to the fiber axis and to the incident beam axis within an angular range delta omega = +/- 35 degrees and recorded the light intensities I kappa. Diffraction efficiencies defined as E kappa = integral of I kappa d omega were studied as a function of sarcomere length and during isometric contraction. The sarcomere length dependences of the efficiencies E kappa of the first three orders show characteristic trends. E1 increases with fiber stretch, E2 has a minimum at a sarcomere length near 2.8 micrometers, and E3 has a maximum near 2.5 micrometers. These trends as well as the observed efficiency ratios are in fairly good agreement with predictions by the intensity formula developed for x-ray structure analysis. During isometric contraction, the diffraction efficiencies of the fiber decrease, with the decreases becoming greater the higher the order number. These decreases might be caused by a longitudinal displacement of myofibrils of up to 0.4 micrometers. The efficiency of light diffraction strongly depends on the tonicity of the bathing fluid. Hypertonic (3/2 x normal) solution reduces E1 to less than half, hypotonic (2/3 x normal) solution increases E1 to almost twice the value obtained in normal Ringer's solution.     

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.     

Real-time measurement of the length of a single sarcomere in rat ventricular myocytes: a novel analysis with quantum dots

As the dynamic properties of cardiac sarcomeres are markedly changed in response to a length change of even ～0.1 μm, it is imperative to quantitatively measure sarcomere length (SL). Here we show a novel system using quantum dots (QDs) that enables a real-time measurement of the length of a single sarcomere in cardiomyocytes. First, QDs were conjugated with anti-α-actinin antibody and applied to the sarcomeric Z disks in isolated skinned cardiomyocytes of the rat. At partial activation, spontaneous sarcomeric oscillations (SPOC) occurred, and QDs provided a quantitative measurement of the length of a single sarcomere over the broad range (i.e., from ～1.7 to ～2.3 μm). It was found that the SPOC amplitude was inversely related to SL, but the period showed no correlation with SL. We then treated intact cardiomyocytes with the mixture of the antibody-QDs and FuGENE HD, and visualized the movement of the Z lines/T tubules. At a low frequency of 1 Hz, the cycle of the motion of a single sarcomere consisted of fast shortening followed by slow relengthening. However, an increase in stimulation frequency to 3-5 Hz caused a phase shift of shortening and relengthening due to acceleration of relengthening, and the waveform became similar to that observed during SPOC. Finally, the anti-α-actinin antibody-QDs were transfected from the surface of the beating heart in vivo. The striated patterns with ～1.96-μm intervals were observed after perfusion under fluorescence microscopy, and an electron microscopic observation confirmed the presence of QDs in and around the T tubules and Z disks, but primarily in the T tubules, within the first layer of cardiomyocytes of the left ventricular wall. Therefore, QDs are a useful tool to quantitatively analyze the movement of single sarcomeres in cardiomyocytes, under various experimental settings.     

Spontaneous and propagated contractions in rat cardiac trabeculae

Sarcomere length measurement by microscopic and laser diffraction techniques in trabeculae of rat heart, superfused with Krebs-Henseleit solution at 21 degrees C, showed spontaneous local sarcomere shortening after electrically stimulated twitches. The contractions originated in a region of several hundred micrometers throughout the width of the muscle close to the end of the preparation that was damaged by dissection. The contractions propagated at a constant velocity along the trabeculae. The velocity of propagation increased from 0 to 10 mm/s in proportion to the number of stimuli (3-30) in a train of electrically evoked twitches at 2 Hz and at an external calcium ion concentration ([Ca++]o) of 1.5 mM. At a constant number of stimuli (n), the velocity of propagation increased from 0 to 15 mm/s with [Ca++]o increasing from 1 to 7 mM. In addition, increase of n and [Ca++]o led to an increase of the extent of local sarcomere shortening during the spontaneous contractions, and the occurrence of multiple contractions. Spontaneous contractions with much internal shortening and a high velocity of propagation frequently induced spontaneous synchronized contractions and eventually arrhythmias. Propagation of spontaneous contractions at low and variable velocity is consistent with the hypothesis that calcium leakage into damaged cells causes spontaneous calcium release from the overloaded sarcoplasmic reticulum in the damaged cells. This process propagates as a result of diffusion of calcium into adjacent cells, which triggers calcium release from their sarcoplasmic reticulum. We postulate that the propagation velocity depends on the intracellular calcium ion concentration, with increases with n and [Ca++]o.     

Myoseverin disrupts sarcomeric organization in myocytes: an effect independent of microtubule assembly inhibition

Although disruption of the microtubule (MT) array inhibits myogenesis in myocytes, the relationship between the assembly of microtubules (MT) and the organization of the contractile filaments is not clearly defined. We now report that the assembly of mature myofibrils in hypertrophic cardiac myocytes is disrupted by myoseverin, a compound previously shown to perturb the MT array in skeletal muscle cells. Myoseverin treated cardiac myocytes showed disruptions of the striated Z-bands containing alpha-actinin and desmin and the localization of tropomyosin, titin and myosin on mature sarcomeric filaments. In contrast, MT depolymerization by nocodazole did not perturb sarcomeric filaments. Similarly, expression of constitutively active stathmin as a non-chemical molecular method of MT depolymerization did not prevent sarcomere assembly. The extent of MT destabilization by myoseverin and nocodazole were comparable. Thus, the effect of myoseverin on sarcomere assembly was independent of its capacity for MT inhibition. Furthermore, we found that upon removal of myoseverin, sarcomeres reformed in the absence of an intact MT network. Sarcomere formation in cardiac myocytes therefore, does not appear to require an intact MT network and thus we conclude that a functional MT array appears to be dispensable for myofibrillogenesis.     

Length-tension relation in Limulus striated muscle

Laser diffraction techniques coupled with simultaneous tension measurements were used to determine the length-tension relation in intact, small (0.5-mm thick, 10-mm wide, 20-25-mm long) bundles of a Limulus (horseshoe crab) striated muscle, the telson levator muscle. This muscle differs from the model vertebrate systems in that the thick filaments are not of a constant length, but shorten from 4.9 to approximately 2.0 micrometers as the sarcomeres shorten from 7 to 3 micrometers. In the Limulus muscle, the length-tension relation plateaued to an average maximum tension of 0.34 N/mm2 at a sarcomere length of 6.5 micrometers (Lo) to 8.0 micrometers. In the sarcomere length range from 3.8 to 12.5 micrometers, the muscle developed 50% or more of the maximum tension. When the sarcomere lengths are normalized (expressed as L/Lo) and the Limulus data are compared to those from frog muscle, it is apparent that Limulus muscle develops tension over a relatively greater range of sarcomere lengths.     

Overextended sarcomeres regain filament overlap following stretch

Sarcomere overextension has been widely implicated in stretch-induced muscle injury. Yet, sarcomere overextensions are typically inferred based on indirect evidence obtained in muscle and fibre preparations, where individual sarcomeres cannot be observed during dynamic contractions. Therefore, it remains unclear whether sarcomere overextensions are permanent following injury-inducing stretch-shortening cycles, and thus, if they can explain stretch-induced force loss. We tested the hypothesis that overextended sarcomeres can regain filament overlap in isolated myofibrils from rabbit psoas muscles. Maximally activated myofibrils (n=13) were stretched from an average sarcomere length of 2.6±0.04μm by 0.9μm sarcomere(-1) at a speed of 0.1μm sarcomere(-1)s(-1) and immediately returned to the starting lengths at the same speed (sarcomere strain=34.1±2.3%). Myofibrils were then allowed to contract isometrically at the starting lengths (2.6μm) for ～30s before relaxing. Force and individual sarcomere lengths were measured continuously. Out of the 182 sarcomeres, 35 sarcomeres were overextended at the peak of stretch, out of which 26 regained filament overlap in the shortening phase while 9 (～5%) remained overextended. About 35% of the sarcomeres with initial lengths on the descending limb of the force-length relationship and ～2% of the sarcomeres with shorter initial lengths were overextended. These findings provide first ever direct evidence that overextended sarcomeres can regain filament overlap in the shortening phase following stretch, and that the likelihood of overextension is higher for sarcomeres residing initially on the descending limb.     

Theoretical Fraunhofer light diffraction patterns calculated from three-dimensional sarcomere arrays imaged from isolated cardiac cells at rest.

Sarcomere striation positions have been obtained throughout the volumes of calcium-tolerant resting heart cells by direct computer interfaced high-resolution optical imaging. Each sarcomere position is stored in a three-dimensional (3-D) matrix array from which Fraunhofer light diffraction patterns have been calculated using numerical methods based on Fourier transforms. Diffraction patterns have been calculated from heart cell data arrays oriented normal to a theoretical laser beam. Twelve characteristic features have been identified and described from these diffraction patterns that correlate to diffraction phenomena observed from both cardiac and skeletal muscle. This numerical approach provides the means to directly assess diffraction pattern formulation, the precision of layer line angular separation, layer-line intensity and angular asymmetries, line widths and fine structures in terms of the known diffracting source structures. These results confirm that theoretical calculations can predict real muscle diffraction patterns and their asymmetries.     

Measurement of sarcomere length during fast contraction of muscle fibers by digital image analysis.

A low-cost, high-resolution (spatial and temporal) image analysis system was developed to measure sarcomere length (Sl) during fast twitch of isolated striated muscle fibers at different temperatures. Fiber images were examined during twitch with an imaging rate of 220 Hz. To increase temporal resolution beyond 220 Hz, consecutive temporally shifted image sequences (N sequences) were acquired. Individual or average Sl was directly measured from a horizontal profile without spatial-frequency assessment. Measurement precision (E) was determined and expressed as: E(%) = 100xPs/(IsxSl), where Ps is the pixel size and Is the involved sarcomere number. At 18 degrees C during isometric twitch, Sls were measured with 220 Hz temporal and 0.2% spatial resolutions. Sl shortened in the central region (0.21+/-0.12 microm) as tension developed, reaching a maximal shortening of 8.09 + 2.05% (at rest, Sl = 2.59+/-0.05 microm, n = 4) in 32.5+/-1.96 ms. At 30 degrees C, Sl variations were examined with 880 Hz temporal resolution, in which case maximal S1 shortening was reached in 15.74+/-1.99 ms, and then decreased to 5.19+/-1.97% (at rest, S1 = 2.6+/-0.06 microm). The twitch tension developed by the whole fiber was recorded and compared with sarcomere length behavior. Sarcomere length variations in the central region were representative of overall developed tensions at 18 and 30 degrees C.     

Active and passive electrical properties of single bullfrog atrial cells

Single cells from the bullfrog (Rana catesbeiana) atrium have been prepared by using a modification of the enzymatic dispersion procedure described by Bagby et al. (1971. Nature [Long.]. 234:351--352) and Fay and Delise (1973. Proc. Natl. Acad. Sci. U.S.A. 70:641--645). Visualization of relaxed cells via phase-contrast or Nomarski optics (magnification, 400--600) indicates that cells range between 150 and 350 micrometers in length and 4 and 7 micrometers in diameter. The mean sarcomere length in relaxed, quiescent atrial cells in 2.05 micrometer. Conventional electrophysiological measurements have been made. In normal Ringer's solution (2.5 mM K+, 2.5 mM Ca++) acceptable cells have stable resting potentials of about -88 mV, and large (125 mV) long- duration (approximately 720 ms) action potentials can be elicited. The Vm vs. log[K+]0 relation obtained from isolated cells is similar to that of the intact atrium. The depolarizing phase of the action potential of isolated atrial myocytes exhibits two pharmacologically separable components: tetrodotoxin (10(-6) g/ml) markedly suppresses the initial regenerative depolarization, whereas verapamil (3 x 10(-6) M) inhibits the secondary depolarization and reduce the plateau height. A bridge circuit was used to estimate the input resistance (220 +/- 7 M omega) and time constant 20 +/- 7 ms) of these cells. Two- microelectrode experiments have revealed small differences in the electrotonic potentials recorded simultaneously at two different sites within a single cell. The equations for a linear, short cable were used to calculate the electrical constants of relaxed, single atrial cells: lambda = 921.3 +/- 29.5 micrometers; Ri = 118.1 +/- 24.5 omega cm; Rm = 7.9 +/- 1.2 x 10(3) omega cm2; Cm = 2.2 +/- 0.3 mu Fcm-2. These results and the atrial cell morphology suggest that this preparation may be particularly suitable for voltage-clamp studies.     

Fine structure in near-field and far-field laser diffraction patterns from skeletal muscle fibers.

Regions of muscle fibers that are many sarcomeres in length and uniform with regard to striation spacing, curvature, and tilt have been observed by light microscopy. We have investigated the possibility that these sarcomere domains can explain the fine structure in optical diffraction patterns of skeletal muscle fibers. We studied near-field and far-field diffraction patterns with respect to fiber translation and to masking of the laser beam. The position of diffracted light in the near-field pattern depends on sarcomere length and position of the diffracting regions within the laser beam. When a muscle fiber was translated longitudinally through a fixed laser beam, the fine structural lines in the near-field diffraction pattern moved in the same direction and by the same amount as the fiber movement. Translation of the muscle fiber did not result in fine structure movement in the far-field pattern. As the laser beam was incrementally masked from one side, some fine structural lines in both the near-field and far-field diffraction patterns changed in intensity while others remained the same. Eventually, all the fine structural lines broadened and decreased in intensity. Often a fine structural line increased in intensity or a dark area in the diffraction pattern became brighter as the laser beam was restricted. From these results we conclude that the fine structure in the laser diffraction pattern is due to localized and relatively uniform regions of sarcomeres (domains) and to cross interference among light rays scattered by different domains.     

Contraction in voltage-clamped, internally perfused single heart cells

We studied contraction in single voltage-clamped, internally perfused myocytes isolated from guinea pig ventricles. The microscopic appearance of the cell was observed and recorded with a television system, while contractile shortening was measured 1,000 times/s using a linear photodiode array. Uniform, synchronous sarcomere shortening occurred in response to depolarizations that triggered a slow inward current (Isi). Changes in Isi caused by altering the amplitude of the voltage step, the extracellular [Ca2+], or the holding potential were accompanied by immediate parallel changes in the extent and velocity of shortening. In particular, twitch shortening during depolarization was immediately decreased when large voltage steps decreased Isi, and was eliminated by depolarizations that exceeded +75 mV, the apparent reversal potential for Ca2+. In these cases, shortening was associated with the tail current during repolarization. Increases in the amplitude, duration, and the rate of the depolarizing step increased the extent and speed of sarcomere shortening over the course of four to five contractions without a simultaneous parallel increase of Isi. Large prolonged depolarizations caused an asynchronous, nonuniform, oscillatory shortening of the cell and potentiated future twitch contractions. Increases in the duration of the depolarizing step immediately prolonged contraction; otherwise, interventions that altered the extent, velocity, and time course of shortening in intact, nonperfused cells did not affect the time course of the contraction in the internally perfused single cells. Our results provide direct support for the hypothesis that Isi both induces and grades the size of the Ca2+ release from the sarcoplasmic reticulum of intact cardiac muscle. In addition, a separate, depolarization-dependent process unrelated to Isi grades the size of contraction, presumably by modulating Ca2+ accumulation in the intracellular stores, and affects its time course.     

Sarcomere length dependence of the force-velocity relation in single frog muscle fibers.

The force-velocity relation of single frog fibers was measured at sarcomere lengths of 2.15, 2.65, and 3.15 microns. Sarcomere length was obtained on-line with a system that measures the distance between two markers attached to the surface of the fiber, approximately 800 microns apart. Maximal shortening velocity, determined by extrapolating the Hill equation, was similar at the three sarcomere lengths: 6.5, 6.0, and 5.7 microns/s at sarcomere lengths of 2.15, 2.65, and 3.15 microns, respectively. For loads not close to zero the shortening velocity decreased with increasing sarcomere length. This was the case when force was expressed as a percentage of the maximal force at optimal fiber length or as a percentage of the sarcomere-isometric force at the respective sarcomere lengths. The force-velocity relation was discontinuous around zero velocity: load clamps above the level that kept sarcomeres isometric resulted in stretch that was much slower than when the load was decreased below isometric by a similar amount. We fitted the force-velocity relation for slow shortening (less than 600 nm/s) and for slow stretch (less than 200 nm/s) with linear regression lines. At a sarcomere length of 2.15 microns the slopes of these lines was 8.6 times higher for shortening than for stretch. At 2.65 and 3.15 microns the values were 21.8 and 14.1, respectively. At a sarcomere length of 2.15 microm, the velocity of stretch abruptly increased at loads that were 160-170% of the sarcomere isometric load, i.e., the muscle yielded. However, at a sarcomere length of 2.65 and 3.15 microm yield was absent at such loads. Even the highest loads tested (260%) resulted in only slow stretch.(ABSTRACT TRUNCATED AT 250 WORDS)     

Individual sarcomere length determination from isolated cardiac cells using high-resolution optical microscopy and digital image processing.

Discrete sarcomere lengths have been determined from dynamically contracting isolated cardiac cells with a high-speed, high-resolution direct optical imaging system. Calcium-tolerant cardiac cells from the rat are isolated by perfusion with collagenase and hyaluronidase. Individual sarcomere lengths can be determined by directly imaging the cell's striation pattern onto a solid-state charge-coupled device (CCD) detector interfaced with a digital computer. The precision of detection in a real light microscopic optical system is discussed in relation to the type of image detector, optical contract enhancement techniques, and digital image processing. The optical performance of the direct striation pattern image apparatus has been determined empirically with test grids under standard bright-field and Nomarski-differential interference contrast (DIC) conditions for application to real muscle imaging. Discrete striation positions of isolated cells have been detected and followed with high precision during phasic contraction-relaxation cycles down to average sarcomere lengths as short as 1.43 +/- 0.053 microns. The maximum rates of contraction and relaxation are rapid and synchronous in time course along the length of the cell. These results indicate that direct optical imaging can provide an accurate means to monitor discrete striations and sarcomere lengths along the length of Ca2+-tolerant heart cells.