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M Tarr

Publications and source records attributed to M Tarr.

At least 37 records · Page 2Linked to original sources

Depression of contractility following stretches and releases applied during contraction to single frog atrial cardiac cells.

The effects of stretches and releases on the contractile performance of isolated single frog atrial cells (Rana catesbeiana) were investigated. A stretch or release was imposed on the cell--either during a contraction (test) or before the onset of contraction (control)--and the contractile performance (length, velocity and force) of the test contraction was compared with that of the control contraction to determine whether the stretch or release imposed on the contracting cell altered the contractility of the cell. We found that the velocity of cell (and sarcomere) shortening for the remainder of the test contraction following either a stretch or release was markedly less than that occurring at the same time in the control contraction. This decreased velocity occurred even though the force in the test contraction was less than that in the control contraction and the sarcomere length was longer in the test contraction than in the control contraction. These results indicate that after a stretch or release imposed on the contracting cell, the force-velocity relationship at any given length and time is depressed than had the stretch or release not been imposed on the contracting cell. Thus, stretches and releases applied to the contracting single cardiac cell either produce a long-term depression in the contractility of the cell, or that the contractility at any given time and sarcomere length depends markedly on the history of the contraction.

Animals↗

Mechanical and contractile properties of isolated single intact cardiac cells.

In light of the demonstrated contractile and mechanical non-uniformities which exist in many of the commonly used intact cardiac preparations, there is now a recognized need to evaluate cardiac sarcomere performance directly in relatively simple cardiac preparations. The single cardiac cell represents the simplest intact cardiac preparation and it can provide the opportunity for relatively definitive studies of cardiac sarcomere performance, since the force at the sarcomere level is fairly well defined compared to the situation in more complex intact cardiac preparations and sarcomere lengths can be measured directly. Considerable progress has been made within the last five years in developing the methodology for preparing intact isolated single cardiac cells and for investigating the mechanical and contractile properties of these cells. There is now sufficient information available to warrant the conclusion that the isolated cardiac cell can have normal force generating and sarcomere shortening capacity as well as normal electrical activity. Studies on single cardiac cells have already given insight into the characteristics of 1) sarcomere shortening and extension at essentially zero external force, 2) sarcomere shortening during auxotonic force development, 3) the contribution which the cell makes to the resting tension-sarcomere length relationship of intact cardiac tissue, and 4) the sarcomere length-dependency of the force generating capacity of cardiac muscle during twitch contractions. These studies have also clearly demonstrated that the cardiac sarcomere has some type of a force-velocity relationship although the exact nature of this relationship remains to be determined.

Animals↗

Effect of external force on relaxation kinetics in single frog atrial cardiac cells.

The effects of external force on relaxation kinetics were investigated in isolated single frog (Rana catesbeiana) atrial cells. We found that force decay occurred at a maximum and constant rate for a significant portion of auxotonic relaxation, and this rate was linearly related to the peak force developed during auxotonic contraction. The slope of the linear relationship between the maximum rate of auxotonic force decay and peak auxotonic force was not affected by changes in the level of contractile activation produced by activating the cell with different stimulus durations. The rate of force change during auxotonic contraction and relaxation in the isolated cell is directly related to the average sarcomere velocity within the cell. Thus, the results indicate that during auxotonic relaxation the velocity of sarcomere extension is directly related to the peak auxotonic force, and sarcomere extension, during relaxation, is therefore affected by external force. The direct effect of external force on relaxation kinetics was confirmed by the observation that force changes imposed on the cell during relaxation immediately altered the velocity of the extending cell from any given length. However, data are also presented which demonstrate that rapid sarcomere extension occurs during relaxation under conditions where external forces are negligible. Thus, rapid sarcomere extension during relaxation does not require large external forces, and internal forces must play a role in sarcomere extension during relaxation. An explanation is given for these apparently contradictory results.

Animals↗

Transmembrane electrical potential differences in cells of isolated renal tubules.

Isolated fragments of renal tubules were prepared by treating rabbit kidneys removed from animals on a normal diet with proteolytic enzymes. The intracellular potential of tubule cells was measured either by impaling cells across the luminal membrane at the everted end of the tubule or by impaling cells across the basolateral membrane along the shaft of the tubule. The mean intracellular potential recorded from 163 cells at the everted ends of proximal tubules was -49 +/- 12 mV (SD) compared with a mean value of -29 +/- 9 mV (SD) recorded from 199 cells along the shaft of the tubules. In papillary collecting ducts there was no significant difference between the intracellular potential of cells at the everted end of the tubules compared with those of cells along the shaft of the tubule. However, two distinctly different types of intracellular potentials were recorded from collecting duct cells regardless of the location of the cells: 93 cell impalements gave a mean intracellular potential of -15 +/- 8 mV (SD), whereas 135 cell impalements gave a mean potential of +24 +/- 12 mV (SD). The data presented in this paper demonstrate that intracellular potentials can be recorded from isolated renal tubule fragments of papillary collecting ducts and proximal convoluted tubules.

Animals↗

Characteristics of sarcomere shortening in single frog atrial cardiac cells during lightly loaded contractions.

We studied sarcomere performance in single isolated intact cardiac cells using techniques that allow direct measurement of sarcomere length and force. This investigation dealt primarily with sarcomere performance during twitch contractions under lightly loaded conditions. In such contractions, there was a significant portion of the contraction in which sarcomere shortening occurred at constant velocity over a significant range of sarcomere lengths. The constant velocity phase of shortening was followed by a phase of shortening in which sarcomere velocity decreased markedly. Both the velocity and extent of sarcomere shortening depended on the stimulus parameters used to excite the cell. With threshold stimulation, sarcomere velocities during the constant velocity phase of shortening ranged from 1 to 5.5 micron/sec in different cells and significant slowing of sarcomere shortening began at sarcomere lengths of 1.8-2.0 micron. In contrast, when cells were stimulated with a long duration stimulus (200 msec) of large current strength, sarcomere velocities during the constant velocity phase ranged from 6 to 12 micron/sec, and significant slowing did not occur until a sarcomere length of about 1.6 micron was reached. The threshold stimulus strength-stimulus duration relationship was determined on the single cell, and it was found to be of the type expected for a cell having an intact excitable membrane capable of generating an action potential when depolarized to a fixed voltage threshold. The data presented in this paper give direct evidence that the lightly loaded cardiac sarcomere has a velocity of shortening which depends on the level of contractile activation but is independent of sarcomere length at sarcomere lengths greater than about 1.6 micron.

Animals↗

Evidence that the velocity of sarcomere shortening in single frog atrial cardiac cells is load dependent.

Recent experiments using laser diffraction techniques to determine the time course and extent of sarcomere shortening in thin bundles of cardial tissue have given results which suggest that the velocity of sarcomere shortening in cardiac muscle is independent of the developed force (Nassar et al., 1974; Krueger and Pollack, 1975). However, the anatomical complexity of the intact tissue precludes a definite interpretation of the data, since the exact relationship between the force being borne by the total tissue to the force being borne by any observed group of sarcomeres is uncertain. The single frog atrial cell provides a simple cardiac preparation in which the relationship between sarcomere velocity and sarcomere force is well defined, since these cells are only 1-2 myofibrils wide. The purpose of the present investigation was to determine if sarcomere velocity in the single frog atrial cell is dependent on force by measuring the time course of sarcomere shortening in single cells under conditions in which the cell developed markedly different forces. The results presented in this paper give direct evidence that the velocity of sarcomere shortening in the single cardiac cell depends on the force being developed by the sarcomeres. Thus, cardiac sarcomeres have a type of force-velocity relationship, although the exact nature of this relationship could not be determined in these experiments.

Animals↗

Effect of initial sarcomere length on sarcomere kinetics and force development in single frog atrial cardiac cells.

We studied sarcomere performance in single isolated intact frog atrial cells using techniques that allow direct measurement of sarcomere length and force. The purpose of this investigation was to determine whether length-dependent alterations in contractile activation occur in the single isolated cardiac cell. This was accomplished by determining the effect of initial sarcomere length on the time course of sarcomere shortening and force development during auxotonic twitch contractions. The results presented in this paper demonstrate that the velocity of sarcomere shortening, the rate of force development, and the magnitude of force development during auxotonic twitch contractions all increase as initial sarcomere length increases over the range of about 2 micrometers to greater than 3 micrometers. These results indicate that the level of contractile activation increases as initial sarcomere length increases. Also, results are presented that indicate that the rate of increase of contractile activation during a twitch contraction also increases as initial sarcomere length increases. These length-dependent effects on contractile activation in conjunction with the slow time course of contractile activation cause the force-velocity-length relationship to be time-dependent: i.e., the velocity of sarcomere shortening at a given sarcomere length and load depends on the time during the contraction when the sarcomere reaches that length. The results suggest that length-dependent alterations in contractile activation may play a major role in the improved contractile performance that accompanies an increase in initial sarcomere length in cardiac muscle.

Animals↗

Activity of carbonic anhydrase in stored blood.

The activity of carbonic anhydrase in blood stored in CPD at 4 degrees C for up to 31 days was measured electrometrically at 5-day intervals and compared with that in fresh blood taken from four normal subjects. No change in activity in stored red cell lysates was observed over this period. There was a clear increase in activity in serum from stored blood with time. These results are discussed in the context of alveolar-to-arterial carbon dioxide gradients seen occasionally in patients following massive transfusions of stored blood.

Blood Preservation↗

Sarcomere length-resting tension relation in single frog atrial cardiac cells.

It generally has been thought that the relatively high resting tension characteristic of cardiac tissue resides in structures (collagen, elastin) external to the individual cardiac cells, but the evidence to support this conclusion has been indirect, since the resting tension of intact single cardiac cells has not been determined previously. The purpose of the present investigation was to determine the resting tension (stress)-sarcomere length relationships of single intact frog atrial cells. For tension determinations, a single cell was attached between two poly-L-lysine coated glass beams; one beam served as a compliant calibrated cantilevered force beam, and length changes were imposed on the cell by movement of the other beam. Coventional bright-field light microscope techniques were used to view the cell, the sarcomere pattern within the cell, and the position of the force beam. The resting tension of the intact cell increased from a value of about 10 nN at a sarcomere length of 2.35 microns to a value of about 130 nN at a sarcomere length of 3.45 microns. Lagrangian and Eulerian resting stress-sarcomere length relationships were computed from the resting tension-sarcomere length relationships. The Lagrangian stress increased from a value of about 0.6 mN/mm2 at a sarcomere length of 2.35 microns to a value of about 7 mN/mm2 at a sarcomere length of 3.45 microns. These values of stress are about 8- to 30-fold less than those previously reported for intact frog atrial tissue and indicate that the resting tension of intact frog atrial preparations resides primarily in structures external to the individual cardiac cell.

Animals↗

Preparation of isolated single cardiac cells from adult frog atrial tissue.

Isolated cardiac cells from bullfrog atrial tissue can be readily prepared by digestion of intact fragments of atrial tissue with trypsin and collagenase. These isolated cells have dimensions of about 5 mum in width and range in length from 300 mum to over 500 mum. Such isolated cells may prove useful for the investigation of contractile activity of cardiac muscle at the single cell level and at the sarcomere level within the single cell.

Animals↗

Limitations of the double sucrose gap voltage clamp technique in tension-voltage determinations on frog atrial muscle.

The purpose of this study was to evaluate the limitations of the double sucrose gap voltage clamp technique in the determination of tension-voltage relationships for frog atrial muscle. Tension-voltage relationships were determined under two conditions. In one case we determined both the tension response and slow inward current associated with an apparent step depolarization (step-clamp) as a function of the magnitude of the step depolarization. In the second case, an action potential was elicited, the voltage clamp was applied early during the plateau phase of the action potential, and the tension response was determined as a function of the clamp potential (action potential-clamp). Under both step-clamp and action potential-clamp conditions, the waveform of the tension response rose to a peak value (Tp) and then decayed with time to a tension that was maintained for the duration of the depolarization. The Tp-clamp potential relationships obtained under step-clamp and action potential-clamp conditions were similar. Microelectrode measurements of transmembrane potential of cells in the "voltage-clamped" region of the preparation demonstrated the lack of temporal and spatial voltage control under both step-clamp and action potential-clamp conditions, and also demonstrated that acquisition of spatial voltage control occurred at about the same time that the tension response reached its peak value. These data indicate that this voltage clamp technique does not allow an accurate determination of the so-called phasic tension-voltage relationship in frog atrial muscle because of a lack of temporal and spatial control of voltage during the rising phase of the tension response.

Action Potentials↗

An assessment of the double sucrose-gap voltage clamp technique as applied to frog atrial muscle.

The homogeneity of voltage clamp control in small bundles of frog atrial tissue under double sucrose-gap voltage clamp conditions was assessed by intracellular microelectrode potential measurements from cells in the test node region. The microelectrode potential measurements demonstrated that (1) good voltage control of the impaled cell existed in the absence of the excitatory inward currents (e.g., during small depolarizing clamp pulses of 10-15 mV), (2) voltage control of the impaled cell was lost during either the fast or slow excitatory inward currents, and (3) voltage control of the impaled cell was regained following the inward excitatory currents. Under nonvoltage clamp conditions the transgap recorded action potential had a magnitude and waveform similar to the intracellular microelectrode recorded action potentials from cells in the test node. Transgap impedance measured with a sine-wave voltage of 1,000 Hz was about 63% of that measured either by a sine-wave voltage of 10 Hz or by an action potential method used to determine the longitudinal resistance through the sucrose-gap region. The action potential data in conjunction with the impedance data indicate that the extracellular resistance (R(s)) through the sucrose gap is very large with respect to the longitudinal intracellular resistance (R(i)); the frequency dependence of the transgap impedance suggests that at least part of the intracellular resistance is paralleled by a capacitance. The severe loss of spatial voltage control during the excitatory inward current raises serious doubts concerning the use of the double sucrose-gap technique to voltage clamp frog atrial muscle.

Action Potentials↗

Equivalent circuit of frog atrial tissue as determined by voltage clamp-unclamp experiments.

The equivalent circuit that has been used in the analysis of nerve voltage-clamp data is that of the membrane capacity in parallel with the membrane resistance. Voltage-clamp experiments on frog atrial tissue indicate that this circuit will not suffice for this cardiac tissue. The change in membrane current associated with a step change in membrane potential does not show a rapid spike of capacitive current as would be expected for the simple parallel resistance-capacitance network. Rather, there is a step change in current followed by an exponential decay in current with a time constant of about 1 msec. This relatively slow capacitive charging current suggests that there is a resistance in series with the membrane capacity. A possible equivalent circuit is that of a series resistance external to the parallel resistance-capacitance network of the cell membranes. Another possible circuit assumes that the series resistance is an integral part of the cell membrane. The data presented in this paper demonstrate that the equivalent circuit of a bundle of frog atrial muscle is that of an external resistance in series with the cell membranes.

Animals↗

Two inward currents in frog atrial muscle.

The double sucrose-gap voltage-clamp technique was applied to frog atrial tissue to investigate the ionic currents responsible for the action potential in this tissue. Membrane depolarization elicited two distinct components of inward current when the test node was exposed to normal Ringer solution: a fast inward current and a slow inward current. The fast inward current appeared to be carried by sodium ions, since it was rapidly abolished by exposure of the fiber to Na(+)-free solution or tetrodotoxin but persisted on exposure to Ca(++)-free solution. In contrast, in the majority of the preparations the slow inward current appeared to be primarily carried by calcium ions, since it was abolished on exposure of the fiber to Ca(++)-free solution but persisted on exposure to Na(+)-free solution. Action potential data supported the voltage-clamp findings. The normal action potential shows two distinct components in the upstroke phase: an initial rapid phase of depolarization followed by a slower phase of depolarization reaching the peak of the action potential. Abolition of the fast inward current resulted in abolition of the initial rapid phase of depolarization. Abolition of the slow inward current resulted in abolition of the slow phase of depolarization. These data support the hypothesis that two distinct and different ionic mechanisms contribute to the upstroke phase of the action potential in frog atrial tissue.

Action Potentials↗