scholarly journals Studies on conformation of F-actin in muscle fibers in the relaxed state, rigor, and during contraction using fluorescent phalloidin.

1983 ◽  
Vol 97 (6) ◽  
pp. 1663-1667 ◽  
Author(s):  
E Prochniewicz-Nakayama ◽  
T Yanagida ◽  
F Oosawa

F-actin in a glycerinated muscle fiber was specifically labeled with fluorescent phalloidin-(fluorescein isothiocyanate) FITC complex at 1:1 molar ratio. Binding of phalloidin-FITC to F-actin affected neither contraction of the fiber nor its regulation by Ca2+. Comparison of polarized fluorescence from phalloidin-FITC bound to F-actin in the relaxed state, rigor, and during isometric contraction of the fiber revealed that the changes in polarization accompanying activation are quantitatively as well as qualitatively different from those accompanying transition of the fiber from the relaxed state to rigor. The extent of the changes of polarized fluorescence during isometric contraction increased with decreasing ionic strength, in parallel with increase in isometric tension. On the other hand, polarized fluorescence was not affected by addition of ADP or by stretching of the fiber in rigor solution. It is concluded from these observations that conformational changes in F-actin are involved in the process of active tension development.

1968 ◽  
Vol 52 (4) ◽  
pp. 682-713 ◽  
Author(s):  
G. A. Langer ◽  
A. J. Brady

The mechanical responses (active and resting tension, dP/dt, TPT) and ionic exchange characteristics (Ca++, K+, Na+) which follow upon a variation in temperature, rate, and [K+]0 were studied in the rabbit papillary muscle and arterially perfused rabbit interventricular setpum. Abrupt changes in temperature provided a means of separating the contributions of rate of development (intensity) of active state and duration of active state to total active tension development (approximated by isometric tension). Threefold changes in duration of active state with proportional changes in active tension can be induced without evidence for alteration of Ca++, K+, or Na+ exchange. Abrupt cooling produced a moderate (∼15%) increase of dP/dt which suggests an augmentation of active state intensity. Evidence is presented to suggest that this increase of dP/dt is based upon an increase in membrane Ca++ concentration which occurs secondary to inhibition of active Na+ transport. The alterations in ionic exchange and active state produced by variation of temperature are discussed in terms of a five-component control system.


1986 ◽  
Vol 87 (5) ◽  
pp. 761-774 ◽  
Author(s):  
R L Moss ◽  
J D Allen ◽  
M L Greaser

Partial extraction of troponin C (TnC) decreases the Ca2+ sensitivity of tension development in mammalian skinned muscle fibers (Moss, R. L., G. G. Giulian, and M. L. Greaser. 1985. Journal of General Physiology. 86:585), which suggests that Ca2+-activated tension development involves molecular cooperativity within the thin filament. This idea has been investigated further in the present study, in which Ca2+-insensitive activation of skinned fibers from rabbit psoas muscles was achieved by removing a small proportion of total troponin (Tn) complexes. Ca2+-activated isometric tension was measured at pCa values (i.e., -log[Ca2+]) between 6.7 and 4.5: (a) in control fiber segments, (b) in the same fibers after partial removal of Tn, and (c) after recombination of Tn. Tn removal was accomplished using contaminant protease activity found in preparations of LC2 from rabbit soleus muscle, and was quantitated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and scanning densitometry. Partial Tn removal resulted in the development of a Ca2+-insensitive active tension, which varied in amount depending on the duration of the extraction, and concomitant decreases in maximal Ca2+-activated tensions. In addition, the tension-pCa relation was shifted to higher pCa values by as much as 0.3 pCa unit after Tn extraction. Readdition of Tn to the fiber segments resulted in the reduction of tension in the relaxing solution to control values and in the return of the tension-pCa relation to its original position. Thus, continuous Ca2+-insensitive activation of randomly spaced functional groups increased the Ca2+ sensitivity of tension development in the remaining functional groups along the thin filament. In addition, the variation in Ca2+-insensitive active tension as a function of Tn content after extraction suggests that only one-third to one-half of the functional groups within a thin filament need to be activated for complete disinhibition of that filament to be achieved.


1968 ◽  
Vol 52 (3) ◽  
pp. 682-713 ◽  
Author(s):  
G. A. Langer ◽  
A. J. Brady

The mechanical responses (active and resting tension, dP/dt, TPT) and ionic exchange characteristics (Ca++, K+, Na+) which follow upon a variation in temperature, rate, and [K+]o were studied in the rabbit papillary muscle and arterially perfused rabbit interventricular setpum. Abrupt changes in temperature provided a means of separating the contributions of rate of development (intensity) of active state and duration of active state to total active tension development (approximated by isometric tension). Threefold changes in duration of active state with proportional changes in active tension can be induced without evidence for alteration of Ca++, K+, or Na+ exchange. Abrupt cooling produced a moderate (∼15%) increase of dP/dt which suggests an augmentation of active state intensity. Evidence is presented to suggest that this increase of dP/dt is based upon an increase in membrane Ca++ concentration which occurs secondary to inhibition of active Na+ transport. The alterations in ionic exchange and active state produced by variation of temperature are discussed in terms of a five-component control system.


1976 ◽  
Vol 231 (5) ◽  
pp. 1501-1508 ◽  
Author(s):  
MJ Siegman ◽  
TM Butler ◽  
SU Mooers ◽  
RE Davies

Mechanical responses to stretch and length-tension relations were examined in rabbit taenia coli, mesenteric vein, aorta, and myometrium and in guinea pig taenia coli made atonic by incubation in Krebs-bicarbonate solution at 20-22 degrees C. When stretched 10% of the length at which maximum active tension is observed (Lo) in 0.5 s, the muscles showed a transient large force (resistance to stretch) that decayed to a new constant level within minutes (stress relaxation). The resistance to stretch decreased markedly in Ca2+-free [disodium ethylene glycolbis-(beta-aminoethylether)-N,N-tetraacetic acid (EGTA)] Krebs but was restored in normal Krebs solution. Calcium removal did not affect the passive length-tension curve. The absence of Ca2+ did not change the steady-state force maintained by the muscle; thus stretch resistance was not due to tone. Blockade of Ca2+ influx associated with electrical activity with 5-[3,4-dimethoxyphenethyl)methylamino]-2-(3,4,5-trimethoxyphenyl-2-isoprop ylvaleronitrile (D-600) and of Ca2+ release from intracellular sites with thymol (1 mM) completely blocked contraction but did not alter the responses to stretch, thus dissociating the responses to stretch from these processes and tension development. The Ca2+-dependent stress relaxation showed a dependence on muscle length similar to that for active tension development. Except at long muscle lengths, where connective tissue markedly affects length-tension relations, most of the "viscoelasticity" of these smooth muscles is dependent on calcium and may be largely due to the straining of crossbridges that are attached, but not generating a net force, in the resting state.


1979 ◽  
Vol 236 (2) ◽  
pp. H244-H253 ◽  
Author(s):  
G. Cooper

During myocardial tetanus, when activation is maximum and constant, there is a linear relationship throughout contraction between oxygen consumption (MVo2) and the cumulative product of active tension and time (integral of AT). The goal of this study was to determine the relation of MVo2 to integral of AT during isometric myocardial twitch contractions. Ten right ventricular cat papillary muscles were studied in a flow respirometer. MVo2 was determined during contractions unloaded from Lmax to a slack length at successive 100-ms intervals after stimulation. In contrast to the linear relationship observed during tetanus, MVo2/integral of AT varied during twitch contractions: when the muscles were made slack 100 ms after stimulation MVo2/integral of AT was 389 +/- 51 (SE) (nl of O2/mg of dry muscle)/(N of active tension/mm2.s of active tension). This value was 94 +/- 7 at peak active tension and was constant thereafter. There was a continuous increase in cumulative MVo2 as integral of AT increased; before integral of AT began, MVo2 was 0.41 +/- 0.04 (nl of O2/mg)/contraction at Lmax and 0.22 +/- 0.04 at a slack length; at peak isometric tension MVo2 was 1.84 +/- 0.19; for a complete contraction MVo2 was 2.89 +/- 0.25. These data support two concepts 1) activation energy is small and dependent on initial length and tension; and 2) integral of AT is variably energy dependent throughout the entire isometric twitch contraction.


1994 ◽  
Vol 266 (1) ◽  
pp. H68-H78 ◽  
Author(s):  
C. R. Cory ◽  
R. W. Grange ◽  
M. E. Houston

The loss of load-sensitive relaxation observed in the pressure-overloaded heart may reflect a strategy of slowed cytosolic Ca2+ uptake to yield a prolongation of the active state of the muscle and a decrease in cellular energy expenditure. A decrease in the potential of the sarcoplasmic reticulum (SR) to resequester cytosolic Ca2+ during diastole could contribute to this attenuated load sensitivity. To test this hypothesis, both in vitro mechanical function of anterior papillary muscles and the SR Ca2+ sequestration potential of female guinea pig left ventricle were compared in cardiac hypertrophy (Hyp) and sham-operated (Sham) groups. Twenty-one days of pressure overload induced by coarctation of the suprarenal, subdiaphragmatic aorta resulted in a 36% increase in left ventricular mass in the Hyp. Peak isometric tension, the rate of isometric tension development, and the maximal rates of isometric and isotonic relaxation were significantly reduced in Hyp. Load-sensitive relaxation were significantly reduced in Hyp. Load-sensitive relaxation quantified by the ratio of a rapid loading to unloading force step in isotonically contracting papillary muscle was reduced 50% in Hyp muscles. Maximum activity of SR Ca(2+)-adenosinetriphosphatase (ATPase) measured under optimal conditions (37 degrees C; saturating Ca2+) was unaltered, but at low free Ca2+ concentrations (0.65 microM), it was decreased by 43% of the Sham response. Bivariate regression analysis revealed a significant (r = 0.84; P = 0.009) relationship between the decrease in SR Ca(2+)-ATPase activity and the loss of load-sensitive relaxation after aortic coarctation. Stimulation of the SR Ca(2+)-ATPase by the catalytic subunit of adenosine 3',5'-cyclic monophosphate-dependent protein kinase resulted in a 2.6-fold increase for Sham but only a 1.6-fold increase for Hyp. Semiquantitative Western blot radioimmunoassays revealed that the changes in SR Ca(2+)-ATPase activity were not due to decreases in the content of the Ca(2+)-ATPase protein or phospholamban. Our data directly implicate a role for decreased SR function in attenuated load sensitivity. A purposeful downregulation of SR Ca2+ uptake likely results from a qualitative rather than a quantitative change in the ATPase and possibly one of its key regulators, phospholamban.


1986 ◽  
Vol 56 (2) ◽  
pp. 494-506 ◽  
Author(s):  
B. R. Botterman ◽  
G. A. Iwamoto ◽  
W. J. Gonyea

Single motor units of the flexor carpi radialis (FCR) muscle were activated with a series of constant-rate stimulus trains to study the relation between the frequency of activation and isometric tension development (F-T relation). The tension produced by each stimulus train was expressed as a percentage of the maximum tension-time area (Amax) found for a given unit. Between 25 and 75% Amax a clear separation was seen in the rates needed to produce the same relative tension for the F-T curves of slow-twitch (type S) and fast-twitch (type F) units. Over the steepest portion of the F-T curve (25-50% Amax), where tension output was most sensitive to changes in activation rate, type F units required substantially higher stimulation rates (30 pps) to achieve the same relative tension output as type S units. Furthermore, the frequency range that corresponded to the steep portion of the curve was 2.3 times greater for type F units. For both type S and F units, twitch duration was deemed to be an important determinant of the F-T curve, as has been shown previously. A direct continuous relation was seen between the integrated twitch time (ITT) and the stimulus interval needed to produce 50% Amax (r = 0.94, P less than 0.001). Thus, units that had relatively brief twitches required higher activation rates to achieve the same relative percentage of Amax. Comparison of F-T curves from FCR with those derived by other investigators for cat hindlimb units (medial gastrocnemius and peroneus longus) revealed that significant differences in activation rates were needed to produce the same percentage of Amax throughout the midrange of the F-T curve. At 50% Amax, type F units in FCR required activation rates approximately 20 pps higher than type F units in the hindlimb. Type S units in FCR required only slightly higher rates (approximately equal to 5 pps). Based on a number of well-founded assumptions, F-T curves derived from FCR units were used to estimate the potential contribution of rate coding to total muscle tension by type S and F units. This analysis leads to the conclusion that rate modulation is a potentially important factor in the gradation of tension for the FCR muscle.


1969 ◽  
Vol 50 (2) ◽  
pp. 431-443 ◽  
Author(s):  
T. HIDAKA ◽  
H. KURIYAMA ◽  
T. YAMAMOTO

1. A study of the mechanical properties of longitudinal muscle in relation to the membrane properties was carried out under isometric conditions. 2. When the stimulus duration exceeded 50 msec., active tension development was followed by sustained contraction. The sustained contractions were not related to spike generation. 3. The critical potassium concentration to produce the contracture depolarized the membrane from -36 to -20 mV. 4. Reduced sodium concentration and increased calcium and increased potassium (up to 27 mM) concentrations enhanced the amplitude of the active tension. 5. Reduced sodium concentration enhanced the amplitude and duration of the sustained tension, but increased potassium and calcium concentrations reduced them. 6. Caffeine (12 mM) induced contractures of the muscle, and reduced the membrane resistance and capacitance. 7. Spikes were not elicited by intracellular stimulation. 8. 5-Hydroxytryptamine (10-5 g./ml.) blocked the generation of the sustained contraction but no effect was observed on the phasic tension.


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