| Stepwise Dynamics of Connecting Filaments Measured in Single Myofibrillar Sarcomeres Biophysical Journal, Volume 74, Issue 3, 1 March 1998, Pages 1473-1483 Paul Yang, Tsukasa Tameyasu and Gerald H. Pollack Abstract Single relaxed myofibrils of bumblebee flight muscle were subjected to motor-imposed ramp-length changes. The image of the striations was projected onto a linear photodiode array, and sarcomere length was computed as the spacing between centroids of contiguous A-bands. Centroid position was determined by integrating the respective A-band intensity peak and computing the location at which the area on one side was equal to the other. The resulting trace of centroid to centroid span versus time was stepwise, with periods of rapid shortening alternating with periods of pause. An alternative nondiscrete sensor gave similar steps. If thick filament length remains constant, stepwise sarcomere length changes imply that length changes in the connecting filament must be stepwise. Thus, shortening of the connecting filament occurs as a sequence of discrete events rather than as a continuous event. Abstract | Full Text | PDF (969 kb) |
| Spontaneous oscillation of tension and sarcomere length in skeletal myofibrils. Microscopic measurement and analysis Biophysical Journal, Volume 61, Issue 5, 1 May 1992, Pages 1099-1108 T. Anazawa, K. Yasuda and S. Ishiwata Abstract We have devised a simple method for measuring tension development of single myofibrils by micromanipulation with a pair of glass micro-needles. The tension was estimated from the deflection of a flexible needle under an inverted phase-contrast microscope equipped with an image processor, so that the tension development is always accompanied by the shortening of the myofibril (auxotonic condition) in the present setup. The advantage of this method is that the measurement of tension (1/30 s for time resolution and about 0.05 micrograms for accuracy of tension measurement; 0.05 microns as a spatial resolution for displacement of the micro-needle) and the observation of sarcomere structure are possible at the same time, and the technique to hold myofibrils, even single myofibrils, is very simple. This method has been applied to study the tension development of glycerinated skeletal myofibrils under the condition where spontaneous oscillation of sarcomeres is induced, i.e., the coexistence of MgATP, MgADP and inorganic phosphate without free Ca2+. Under this condition, we found that the tension of myofibrils spontaneously oscillates accompanied by the oscillation of sarcomere length with a main period of a few seconds; the period was lengthened and shortened with stretch and release of myofibrils. A possible mechanism of the oscillation is discussed. Abstract | PDF (1614 kb) |
| Half-Sarcomere Dynamics in Myofibrils during Activation and Relaxation Studied by Tracking Fluorescent Markers Biophysical Journal, Volume 90, Issue 2, 15 January 2006, Pages 514-530 Ivo A. Telley, Jachen Denoth, Edgar Stüssi, Gabriele Pfitzer and Robert Stehle Abstract To study the dynamics of individual half-sarcomeres in striated muscle contraction, myofibrils prepared from rabbit psoas muscle and left ventricles of guinea pig were immunostained with two conjugated antibody complexes consisting of a primary antibody against either -actinin or myomesin and a secondary fluorescently labeled Fab-fragment. We simultaneously measured force kinetics and determined the positions of the Z-line and M-band signals by fluorescence video microscopy and sophisticated computer vision (tracking) algorithms. Upon calcium activation, sarcomeres and half-sarcomeres shortened nonuniformly. Shortening occurred first rapidly and exponentially during the force rise and then slowly during the force plateau. In psoas myofibrils, time-resolved displacements of the A-band in sarcomeres were observed, i.e., the two halves of individual sarcomeres behaved nonuniformly. Nonuniformity in length changes between the two halves of sarcomeres was comparable to that between two adjacent half-sarcomeres of neighboring sarcomeres. Sequential lengthening of half-sarcomeres was observed in cardiac myofibrils during the rapid phase of force relaxation. The independent dynamics of the halves in a sarcomere reveals the half-sarcomere as the functional unit rather than the structural unit, the sarcomere. The technique will facilitate the study of filament sliding within individual half-sarcomeres and the mechanics of intersegmental chemomechanical coupling in multisegmental striated muscles. Abstract | Full Text | PDF (842 kb) |
Copyright © 1996 The Biophysical Society. All rights reserved.
Biophysical Journal, Volume 71, Issue 5, 2774-2785, 1 November 1996
doi:10.1016/S0006-3495(96)79470-5
Research Article
A.L. Friedman and Y.E. Goldman
Department of Bioengineering, University of Pennsylvania, Philadelphia 19104, USA.
A new instrument, based on a technique described previously, is presented for studying mechanics of micron-scale preparations of two to three myofibrils or single myofibrils from muscle. Forces in the nanonewton to micronewton range are measurable with 0.5-ms time resolution. Programmed quick (200-microseconds) steps or ramp length changes are applied to contracting myofibrils to test their mechanical properties. Individual striations can be monitored during force production and shortening. The active isometric force, force-velocity relationship, and force transients after rapid length steps were obtained from bundles of two to three myofibrils from rabbit psoas muscle. Contrary to some earlier reports on myofibrillar mechanics, these properties are generally similar to expectations from studies on intact and skinned muscle fibers. Our experiments provide strong evidence that the mechanical properties of a fiber result from a simple summation of the myofibrillar force and shortening of independently contracting sarcomeres.