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• Sarcomeric domain organization within single skinned rabbit ...

  Sarcomeric domain organization within single skinned rabbit psoas fibers and its effects on laser light diffraction patterns Biophysical Journal, Volume 48, Issue 6, 1 December 1985, Pages 967-982 B. Brenner Abstract Total intensity and fine structure of first-order laser light diffraction maxima from single skinned rabbit psoas fibers were studied. Total intensity of the diffraction maxima was measured as a function of the incidence angle (omega-scan). In the most homogeneous fibers, most of the intensity in the diffraction maxima is confined to a rather narrow range of incidence angles. Fibers with less homogeneous striation patterns, apparently composed of several regions of distinct sarcomere length and tilt of striation (domains), give rise to several narrow intensity peaks in their omega-scans. Left and right first-order diffraction lines produce omega-scans of almost identical shape, composed of one or more intensity peaks, with each pair of corresponding peaks separated by about the same angle. The data indicated that in single skinned rabbit psoas fibers, light diffraction is dominated by Bragg diffraction and that the peaks within omega-scans can be directly correlated with domains within the illuminated fiber segment. In the most homogeneous fiber segments the diameter of domains, estimated from the width of the corresponding maxima in the omega-scans, could almost be as large as the fiber diameter. On average, from the number of peaks in the omega-scans two to three domains with an average length of approximately 250–350 microns can be identified in a fiber cross-section. Therefore, on average only a small number of domains (8 per mm) are found within skinned rabbit psoas fiber segments. In contrast, the number of substructural lines within the diffraction maxima is large even for microscopically homogeneous fibers. Substructural lines appear to be present only when several domains are illuminated simultaneously. Separation and width of these substructural lines are approximately inversely proportional to the length of the illuminated region of the fiber. These data suggest that the substructural lines are due to interference between domains, illuminated simultaneously by a light source with a high degree of spatial coherence (laser). The relevance of these findings for measurements of sarcomere length by laser light diffraction is discussed. Abstract | PDF (3765 kb)

• Desmin Filaments Influence Myofilament Spacing and Lateral C...

  Desmin Filaments Influence Myofilament Spacing and Lateral Compliance of Slow Skeletal Muscle Fibers Biophysical Journal, Volume 88, Issue 2, 1 February 2005, Pages 1156-1165 J. Balogh, Z. Li, D. Paulin and A. Arner Abstract Intermediate filaments composed of desmin interlink Z-disks and sarcolemma in skeletal muscle. Depletion of desmin results in lower active stress of smooth, cardiac, and skeletal muscles. Structural functions of intermediate filaments in fast (psoas) and slow (soleus) skeletal muscle were examined using x-ray diffraction on permeabilized muscle from desmin-deficient mice (−/−) and controls (+/+). To examine lateral compliance of sarcomeres and cells, filament distances and fiber width were measured during osmotic compression with dextran. Equatorial spacing (x-ray diffraction) of contractile filaments was wider in soleus −/− muscle compared to +/+, showing that desmin is important for maintaining lattice structure. Osmotic lattice compression was similar in −/− and +/+. In width measurements of single fibers and bundles, −/− soleus were more compressed by dextran compared to +/+, showing that intermediate filaments contribute to whole-cell compliance. For psoas fibers, both filament distance and cell compliance were similar in −/− and +/+. We conclude that desmin is important for stabilizing sarcomeres and maintaining cell compliance in slow skeletal muscle. Wider filament spacing in −/− soleus cannot, however, explain the lower active stress, but might influence resistance to stretch, possibly minimizing stretch-induced cell injury. Abstract | Full Text | PDF (161 kb)

• Measurement of sarcomere shortening in skinned fibers from f...

  Measurement of sarcomere shortening in skinned fibers from frog muscle by white light diffraction Biophysical Journal, Volume 52, Issue 1, 1 July 1987, Pages 57-68 Y.E. Goldman Abstract 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. Abstract | PDF (2790 kb)

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Abstract

Light diffraction patterns from isolated frog semitendinosus muscle fibers were examined. When transilluminated by laser light, the muscle striations produce a diffraction pattern consisting of a series of lines that are projected as points onto an optical detector by a lens system. Diffraction data may be sequentially stored every 18 ms for later processing by digital computer systems. First- and second-order diffraction line intensities were examined from intact, chemically skinned, and glycerinated single fibers. The diffraction line intensities demonstrated a strong length dependence upon passive stretch from reference length to 3.6 micrometer. The first-order intensity linearly increased an average of 15-fold over the range examined. The magnitude of the second order intensity was less than the first order and showed an exponential rise with increasing length. Both first- and second-order intensities decreased upon muscle activation. Data from chemically skinned and glycerinated single fibers were not significantly different from intact fibers, indicating that the membrane structure has little effect upon the diffraction phenomenon in muscle. Theoretical model systems are examined in an attempt to find the basis of these results. Neither an analysis based on a diffraction grating with variable spacing nor the unit cell model of Fujime provides an explanation for the observed length dependency of intensity. Though the origin of the intensity decrease upon stimulation is not known, we have suggested that it could result from lateral misalignment of myofibrils and can occur upon activation.