Cross-bridge properties derived from muscle isotonic velocity transients.

The rate constants for the turnover of cross-bridges during frog muscle contraction were determined from an analysis of the motion that follows step decreases in load. For a given projection from the myosin filament, there appears to be a range of about 100 A along the length of the filament over which the projection can attach to the actin filament and form a cross-bridge. The site of attachment is then displaced by a distance of this same order before the link is broken. The values of the rate constants also imply that a cross-bridge is formed each time an actin site comes within range of a myosin projection, so that the turnover of cross-bridges for a given contraction distance is independent of the speed of the motion.

Failure of nitroglycerin introduced after prolonged myocardial ischemia to improve collateral blood flow and function in tranquilized dogs.

This study investigated whether nitroglycerin can improve ischemic zone blood flow and function when its infusion is delayed following left anterior descending (LAD) occlusion. Nitroglycerin (200 micrograms/min, 11 dogs) or saline (six dogs) was infused for 2 hours starting 2 hours after occlusion. Regional myocardial blood flow (MBF) was measured (9 +/- 1 micron radioactive microspheres) before and at 2 and 4 hours after occlusion. Segmental contraction was determined by cineroentgenography of implanted tantalum markers. For all ischemic samples (defined as MBF less than or equal to 0.4 ml/min/gm), the average improvement in MBF in the epicardial half (EPI) was 0.05 +/- 0.02 ml/min/gm (mean +/- SEM) with nitroglycerin vs 0.06 +/- 0.06 with saline (p greater than 0.5). Improvement in the endocardial half (ENDO) averaged 0.03 +/- 0.03 ml/min/gm with nitroglycerin vs 0.09 +/- 0.08 with saline (p = 0.5). Contraction in the ischemic zone ceased following occlusion and was unaffected by nitroglycerin or saline. Control blood flows in the ischemic region were 22% less in the ENDO (p less than 0.001) and 19% less in the EPI (p less than 0.005) than in nonischemic myocardium. These results indicate that 2 hours after LAD occlusion in dogs, nitroglycerin was unable to improve ischemic zone collateral flow or contractile function compared to untreated controls. Lower ischemic zone control flows indicate that infarct volume expansion may occur within 4 hours after coronary occlusion.

Contraction transients of skinned muscle fibers.

The contraction kinetics of calcium-activated skinned muscle fibers were studied after step decreases in load by means of a quickly responding recording system. The steady velocity at a given relative load was close to that found in electrically stimulated, intact muscle fibers. The presteady motion had the same shape as that of intact fibers, but the time scale of the transient was nearly two times slower. The duration of the initial phase of the motion, where the velocity was greater than the steady value, and the time at which the subsequent low velocity phase ended, were both stretched out to the same extent. Changing the temperature had the same effect on the length of these two phases of the transient. The results indicate that both phases of the transient are produced by the same underlying factors and can be taken as evidence that the entire transient originates in the crossbridge mechanism. In this case the experimental technique described here provides a basis for distinguishing between chemical parameters that affect contractility by (a) controlling the number of sites at which crossbridges can be formed, as opposed to (b) changing the kinetic properties of a given number of sites.

Relative magnitude and asynchrony of directional components of contraction in sedated dog left ventricle.

The purpose of this study was to quantitate the temporal relationships and the extent and speed of shortening in segments of myocardium responsive to contraction in circumferential, longitudinal, and oblique fiber groups. Measurements were made in five sedated dogs (morphine, diazepam) with and without alterations in preload and afterload (nitroprusside, phenylephrine). The measurement interval was the phase of rapid contraction, determined by differentiation of the segment length vs. time. In the control state, percentage segment shortening was greater in circumferential than in longitudinal [15.2 +/- 0.24 (SE) vs. 10.5 +/- 0.80%; P = 0.0020] and in the subepicardial oblique than in the subendocardial oblique fiber directions (16.6 +/- 0.65 vs. 9.7 +/- 0.36%; P = 0.0010). Shortening was proportional to both maximum speed and duration of shortening (r = 0.735 +/- 0.015 and 0.757 +/- 0.017, respectively). Duration of shortening was significantly longer in circumferential than in longitudinal (mean difference 39.3 +/- 6.6 ms; P = 0.0039) and in subepicardial oblique than in subendocardial oblique directions (mean difference 27.7 +/- 5.5 ms; P = 0.0072). Velocities of up to 3.0 segment lengths/s were attained in response to nitroprusside. These data reveal the local anisotropy and asynchrony of contraction in the myocardium; however, they also support the concept of the myocardium as a functional continuum. The dominance of circumferential over longitudinal and subepicardial over subendocardial oblique contractile components indicates their relative contributions to the constriction of the midmyocardial shell.

Patterns of cellular injury in myocardial ischemia determined by monoclonal antimyosin.

The development of cellular injury in the rat left ventricle resulting from left coronary artery occlusion was examined by immunofluorescence after intravenous injection of monoclonal antimyosin. Cardiac muscle cells that bound antimyosin during ischemia were localized by staining sections with fluorescein-conjugated anti-mouse IgG. Fluorescent staining was detectable within the ischemic region of the left ventricle 3 hr after occlusion and injection of antimyosin. After 6 hr of ischemia, the highly irregular margin of the ischemic zone was clearly outlined by fluorescent cells. At 3-6 hr after occlusion, marked heterogeneity in cellular staining was observed in the epicardial half of the ischemic area, with intensely fluorescent cells intermixed with cells of markedly lower fluorescence. By 24 hr, a homogeneous pattern of staining was observed throughout the ischemic zone. In nonischemic regions of the heart and in rats treated for 24 hr with antimyosin without occlusion, there were only background levels of staining. We conclude that: (i) visualization of ischemic cells via antimyosin provides a sensitive means for examining developing patterns of injury; (ii) the heterogeneity of staining during early ischemia may reflect variation in cellular resistance to deprivation; and (iii) the pattern of fluorescence at the margin of the occluded region indicates that the "border zone" is composed of interdigitating ischemic and nonischemic tissues.