Biomedical subjects

D A Martyn

Publications and source records attributed to D A Martyn.

28 records · Page 2Linked to original sources

PubMed1992

Force and stiffness in glycerinated rabbit psoas fibers. Effects of calcium and elevated phosphate.

Force (F) and stiffness (K) were measured in glycerinated psoas fibers at various calcium levels with 0, 10, 20, and 30 mM orthophosphate (Pi) added to the bathing solutions. The concentrations of bathing solution constituents were as follows: 110 mM potassium, 40 mM sodium, 4 mM MgATP, 10 mM total EGTA, and variable amounts of MOPS (pH buffer). The pH was 7.0, the ionic strength was 200 mM, and the temperature was 10 degrees C. Calcium levels were established by adding various amounts of CaCl2. All solutions contained 4% Dextran T-500. Fiber K was measured by imposing sinusoidal length changes (0.03-0.1%) at 1 kHz and by applying rapid steps in length and measuring the resulting F changes. At all [Pi] tested, K was more sensitive to calcium than F. Elevating bathing solution [Pi] caused a decrease in the calcium sensitivity of both F and K, while the slopes of F-calcium and K-calcium relations increased. In maximally activating calcium, raising [Pi] caused a continuous decrease in F over the range tested, while from very low to 10 mM Pi K remained constant. Above 10 mM Pi K declined, but to a lesser extent than did F. The results suggest that under our experimental conditions strongly attached crossbridges can exist in both force-producing and non-force-producing states, and that the relative population of these states may be calcium dependent.

Actins↗

PubMed1992

Effects of cycling and rigor crossbridges on the conformation of cardiac troponin C.

The results of work by several investigators indicate that crossbridge attachment serves as a positive feedback mechanism that transiently increases the Ca2+ affinity of troponin C (TnC) during each normal heartbeat. To monitor structural changes in the cardiac isoform of TnC (cTnC) associated with Ca2+ binding and crossbridge attachment in muscle, we labeled cTnC with the sulfhydryl-specific fluorescent probe 2-(4'-iodoacetamidoanilino)naphthalene-6-sulfonic acid (IAANS). When IAANS-labeled cTnC (cTnCIAANS) was substituted for endogenous TnC, the fluorescence intensity of cardiac and skeletal muscle preparations increased substantially during rigor crossbridge attachment in the absence of Ca2+ (pCa 9.2). In cardiac muscle, the fluorescence signal increased the same amount in rigor and maximal activation, whereas in skeletal muscle, it was higher in rigor (rigor: cardiac and skeletal = 1; pCa 4.0: cardiac = 0.98 +/- 0.13, skeletal = 0.59 +/- 0.05). This indicates that crossbridge attachment alone is capable of influencing the structure of cTnCIAANS. Because the relative fluorescence intensity of cTnCIAANS was more sensitive to Ca2+ than was force in both preparations (cardiac: pCa50 fluorescence = 6.05 +/- 0.05, pCa50 force = 5.51 +/- 0.11; skeletal: pCa50 fluorescence = 5.94 +/- 0.13, pCa50 force = 5.65 +/- 0.14), we measured the Ca2+ sensitivity of the strong crossbridge attachment (sinusoidal stiffness was measured by imposing 1 kHz at 0.1-0.2% muscle length) in rat trabeculae.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Length and myofilament spacing-dependent changes in calcium sensitivity of skeletal fibres: effects of pH and ionic strength.

The calcium sensitivity of force was measured in glycerinated rabbit psoas fibres at sarcomere lengths (SL) from 2.3 to 3.4 micron. Increased SL caused calcium sensitivity to increase and the slope of force-calcium relations to decrease. We have hypothesized that length-dependent changes in myofilament lattice spacing and the presence of fixed charge on the myofilaments are important in determining calcium sensitivity. Lattice spacing changes were monitored by measuring fibre diameter (D). D was decreased by increasing SL, decreasing bathing solution pH and by osmotic compression with 3% PVP. 3% PVP caused D to decrease by about 15% at all SLs and pH values tested. Force-calcium relations were measured at different SLs and pH values, with and without 3% PVP in the bathing solutions. At all pH values D at SL 2.3 micron with 3% PVP was comparable to the value at 3.4 micron, without PVP. At pH 7.5 and 7.0 calcium sensitivity was about the same at both SL, although the slope of the force-calcium relation was less at longer SL. The similarity of the calcium sensitivity at the same D, but much different SL, indicates that lattice spacing is important in determining calcium sensitivity, while SL and the degree of myofilament overlap are important in determining the slope of force-calcium relations. In order to test for the role of myofilament charge in determining calcium sensitivity, pH and ionic strength were varied. Decreasing pH caused decreased maximum force and calcium sensitivity. In addition, the influence of SL on calcium sensitivity decreased as pH was lowered, with minimal SL dependence at pH 5.5; even though lattice spacing still decreased with increasing SL. When D was decreased with PVP, calcium sensitivity increased at all SLs in pH 7.5 and 7.0 while the same lattice spacing changes at pH 6.0 and 5.5 resulted in greatly reduced shifts in calcium sensitivity. These results indicate that the effect of lattice spacing on calcium sensitivity depends on myofilament charge. At pH 6.0, even though osmotic compression of the lattice has no effect, increasing SL causes about half the shift in calcium sensitivity seen at pH 7.0. Lowering ionic strength from 200 to 110 mM caused an increase in both the magnitude and length dependence of calcium sensitivity at pH 7.0, while at pH 5.5 both decreased.(ABSTRACT TRUNCATED AT 400 WORDS)

Actin Cytoskeleton↗

PubMed1984

Calcium sensitivity is modified by contraction.

This paper summarizes three lines of experimental evidence showing that crossbridge interaction affects calcium sensitivity and probably also affects calcium binding. Evidence is presented that this is a true hysteresis, not just a slow approach to equilibrium. In barnacle single muscle fibers injected with aequorin to monitor intracellular Ca, a long duration stimulus under voltage clamp conditions can produce a long duration calcium transient and force record which both approach steady levels. However, if the stimulus is briefly elevated to transiently produce a higher force early in the contraction, the same steady state Ca level can eventually maintain a higher steady force. Thus Ca sensitivity is modified. In "skinned" barnacle muscle activated by Ca in the presence of buffered Ca, MgATP, and pH, force was measured in split, detergent treated fiber while it was transferred consecutively between solutions which were relaxing, submaximal contracting, maximal contracting, the same submaximal contracting, and finally relaxing. The submaximal contracting solution produced more force when stepping down in Ca concentration (as in relaxation) than when stepping up. Extending the time in the initial submaximal contracting solution did not result in more force. Thus, the force-pCa relationship shows marked hysteresis. The same phenomenon was seen in frog and mammalian muscle fibers. These experiments confirm the findings that contraction modifies Ca sensitivity. In the barnacle single muscle fiber preparation (under both voltage clamp and controlled length conditions), phasic (400 msec) depolarization leads to a calcium transient and a twitch contraction. Releasing the muscle to allow it to shorten rapidly during the declining phase of the calcium transient causes the force to fall and leads to extra Ca in the sarcoplasm. Rapidly stretching the muscle produces the opposite effect. The extra Ca probably comes from a myofilament Ca activating site. Thus, a length change (force change) affects Ca binding.

Aequorin↗

PubMed1984

The dependence of force and velocity on calcium and length in cardiac muscle segments.

The segment length (SL) dependence of force (F) and light load shortening velocity (VL) was determined for central segments of ferret papillary muscles at different extracellular calcium concentrations. Muscles were maintained at 27 degrees C in a physiological solution which contained in mM: NaCl 140; KCl 5.0; MgSO4 1.0; NaH2PO4 1.0; acetate 20; the pH was 7.4. Calcium concentrations were 1.125, 2.25, 4.5 and 9.0 mM. Total force-segment length relations were determined from both muscle length isometric ( auxotonic ) and segment isometric contractions, and were found to be the same for each contraction mode. The peak force generated at a particular segment length was independent of both the amount of shortening during a contraction and the initial SL. Increasing extracellular Ca2+ shifted the F-SL relation toward greater force and the SL axis intercept toward shorter SL. Maximum peak twitch tension was achieved in 9.0 mM Ca2+. Calcium variations also changed the shape of the total F-SL relation from linear in high Ca2+, to concave in low Ca2+. In order to estimate the active F-SL relations, corrections were made for passive force by two methods. The first assumed that passive force was related to SL, and yielded F-SL relations which were nearly identical to those found for total force. This similarity included the curvature changes observed in different Ca2+ concentrations, a finding which is consistent with the hypothesis that length dependent activation is the cause of force decline at short SL. The second method assumed passive force to be related to muscle length, an approach which would be appropriate if, for example, a connective tissue sheath on the muscle dominated passive behavior. These F-SL curves displayed a plateau above 90% SLmax and appeared to be vertically shifted versions of each other. Such characteristics are consistent with the possible role of an internal load in causing the decline of force at short SL. VL-SL relations were obtained from load clamps to 1 mM, imposed at various times during a segment isometric twitch. The results indicate that 1) VL declines linearly with SL below 90% SLmax and 2) VL-SL relations are shifted to higher velocity and shorter SL axis intercepts by increasing Ca2+. The slopes of the VL-SL relations obtained in different calciums are similar. Although an internal load could explain the calcium dependence of VL, it would not explain the similarity of the slopes of the VL-SL relations found in different calciums .

Animals↗

PubMed1983

Hysteresis in the force-calcium relation in muscle.

Calcium ions activate muscle contraction. The mechanism depends on the calcium sensitivity of the proteins that regulate contraction. Evidence is presented for the reverse phenomenon, where contraction modulates calcium sensitivity. Increasing the force level increased calcium sensitivity in intact fibers showing that the relation between force and calcium is not unique. A particular calcium concentration can maintain a higher force level than it can create. The results were confirmed in skinned fiber experiments. Transient reduction of the force led to a transient reduction in calcium binding, suggesting a simple mechanism for the hysteresis.

Aequorin↗

PubMed1983

Force-length relations in cardiac muscle segments.

Using a new technique that measures the length of a segment in the central region of isolated papillaries, we have determined the force-segment length relation for ferret papillary muscles at 27 degrees C. The muscles contracted under muscle length isometric (auxotonic) and segment isometric conditions in physiological solutions containing 9.0, 4.5, 2.25, and 1.125 mM Ca2+. Force-segment length relations obtained from auxotonic and segment isometric contractions were identical in a given Ca2+ concentration. Calcium variations, however, changed the position, shape, and segment length intercept of the force-segment length relation. Force, at a given segment length, increased with increasing Ca2+ up to 9.0 mM Ca2+. The force-segment length relation changed shape from linear, in 4.5 mM Ca2+, to concave in 1.125 mM Ca2+. The segment length intercept was found by extrapolation to be 68, 69, and 74% SLmax in 4.5, 2.25, and 1.125 mM Ca2+, respectively. Two passive force corrections were used to calculate the developed force-segment length relations. Assuming passive force to be related primarily to segment length yields curves that change shape with Ca2+ concentration, suggesting length-dependent activation. On the other hand, assuming passive force to be related to muscle length results in curves for different Ca2+ concentrations that are nearly vertically shifted versions of each other, suggesting the influence of internal loads.

Animals↗

PubMed1983

Myocardial segment velocity at a low load: time, length, and calcium dependence.

The length and time dependence of shortening velocity (VL) at a very light load (1 mN) was determined for central segments of ferret papillary muscle at 27 degrees C. A recently developed technique that measures the cross-sectional area of the chosen segment was used to assess segment length. Segment length (SL) or force could be used as a feedback control signal. VL was determined by releasing the muscle to a 1-mN load (less than or equal to 3% maximum force) at various times during a segment isometric twitch. Segment isometric twitches were typically performed at 95-98% SLmax, SLmax being the longest segment length attainable in the preparation. VL, as a function of time and SL, was determined from SL shortening during the load clamp. Muscle responses were sampled, stored, and analyzed with an on-line digital computer. The data indicate that 1) at a given SL, VL declines from 20 to 50% of its peak value by the time of peak force production, 2) VL declines linearly with SL below 90% SLmax in 1.125, 2.25, and 4.5 mM extracellular Ca2+, 3) VL exhibits a dependence on extracellular Ca2+, having peak values at 90% SLmax of 3.32 +/- 0.33, 2.66 +/- 0.17, and 1.89 +/- 0.17 SL/s in 4.5, 2.25, and 1.125 mM extracellular Ca2+, respectively.

Animals↗

PubMed1983

Onset of relaxation in cardiac muscle segments.

The onset of relaxation has been studied in undamaged central segments of isolated ferret papillary muscles at 27 degrees C, 12 beats/min. A technique that provides a signal proportional to the length of a chosen segment was used to assess segment velocity and length. Feedback control was employed to obtain segment isometric contractions. At a variety of times during segment isometric twitches, rapid load clamps were imposed using a range of loads from resting force to greater than half peak developed force. For the purposes of this study, the onset of relaxation was defined as occurring when active segment shortening ceased and elongation began (i.e., Vseg = 0). Early load clamps to low loads resulted in V = 0 at comparatively short segment lengths and early times. Later load clamps caused zero velocity to occur at longer segment lengths and later times. The V = 0 points in fact formed a line in the segment length-time plane. Contractions clamped to higher loads exhibited reduced shortening and a prolonged time course so that the V = 0 points showed the same dependence on length and time. Remarkably, all the variations of load-clamp load, time, and initial length yielded V = 0 points that were intermixed along a single line. Increasing or decreasing extracellular Ca2+ caused the V equal to O points to shift to later times and shorter segment lengths or earlier times and longer segment lengths, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1980

The effects of acute and chronic ethanol administration on twitch and tetanic force production by rat gastrocnemius muscle in situ.

The effects of acute ethanol injection on force production were measured in normal and chronic alcoholic rats. Acutely injected normal rats, acutely injected chronic alcoholic rats, and noninjected chronic alcoholic rats were compared with control groups: normal nonalcoholic rats and a dietetically deprived control group. Acute ethanol injection increased twitch force in normal rats but not in chronic alcoholic rats. Chronic alcohol ingestion caused a decrease in maximal twitch and tetanic tension. The dietetically deprived control group developed less maximal tetanic force, with no significant decrease in maximal twitch force. Chronic alcohol ingestion seemed to decrease maximal twitch tension, whereas the decrease in maximal tetanic could be attributed to the effects of malnutrition.

Alcoholism↗