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Biomedical subjects

B D Lindley

Publications and source records attributed to B D Lindley.

At least 19 recordsLinked to original sources

Activation in frog atrial trabeculae: dependence on temperature and length.

Isolated frog atrial trabeculae were activated using the method of Na+ withdrawal to induce contractures of relatively steady tension. External Na+ concentration [( Na+]o) during contractures was varied between 0.25 and 45 mM. Isometric contracture tension was measured at cold (4 degrees C) and warm (20 degrees C) temperatures. In addition, rapid temperature jumps (complete in approximately 400 ms) were imposed during cold contractures, resulting in tension transients that consisted of an initial increase in tension followed by a decrease, the latter phase being greater at small and moderate reductions in [Na+]o. Peak contracture tension varied with relative muscle length. The trabeculae became more sensitive with stretch to Na+ withdrawal at 20 degrees C and generated relatively greater tensions at a given [Na+]o. The initial tension increase after a temperature jump was directly proportional to the peak contracture tension immediately preceding the increase in temperature and was therefore interpreted as reflecting an effect of the higher temperature on the attached force-generating cross bridges. The effects of cold and warm steady temperatures and temperature jumps during isometric twitches were also studied. Peak twitch tension varied inversely with temperature (stimulus frequency = 0.2 Hz). In contrast, temperature jumps imposed during the rising phase of twitches at a steady cold temperature (approximately 4 degrees C) resulted in a large initial increase in tension followed by relaxation at a rate that was characteristic of the elevated temperature. The results suggest that, at the warmer temperature (approximately 20 degrees C), activation (i.e., number of attached cross bridges) of the myocardium is significantly less than maximal during the twitch response. The dependence of the tension vs. [Na+]o curves and the tension transients resulting from the temperature jumps on relative muscle length provide evidence for a length dependency of contractile activation in intact atrial trabeculae under conditions of steady-state tension development.

Animals

Influence of temperature upon contractile activation and isometric force production in mechanically skinned muscle fibers of the frog.

Increasing temperature (4-22 degrees C) increases the Ca2+ concentration required for activation of mechanically skinned frog muscle fibers. The pCa required for 50% maximal force (pCa50) was inversely proportional to absolute temperature. Assuming that relative force is directly related to fractional occupancy of the Ca2+-binding sites on troponin that regulate force, the shift was consistent with a Gibbs free energy change of binding (delta G) of about -7.8 kcal/mol. This is close to the delta G for Ca2+ binding to the calcium-specific sites on troponin C reported by others. Decreasing Mg2+ from 1 mM to 60 microM shifts the force-pCa curves at either 4 or 22 degrees C to higher pCa, but the shift of pCa50 with temperature over this range (0.4 log units) was the same at low and high Mg2+. Maximal force increased with temperature for the entire range 4-22 degrees C with a Q10 of 1.41, and over the restricted range 4-15 degrees C with a Q10 of 1.20. From the dual effects of temperature on Ca2+ activation and maximal force, one would expect that force would respond differently to temperature change at high or low Ca2+. At high Ca2+, a temperature increase will lead to an increased force. However, at low to intermediate Ca2+ levels (below the intersection of the force-pCa curves for the initial and final temperatures), steady state force should decrease with increasing temperature. The inverse responses should occur with a decrease in temperature. These responses are observed when temperature is changed by rapid solution exchange.

Animals

Studies on the mechanism of fluid secretion by isolated salivary glands of Calliphora.

1. Potassium is the major cation in the secretion of the salivary glands of Calliphora and is necessary for full secretory rates. 2. Other ions (rubidium and sodium) can support secretion in the absence of potassium. 39. During stimulation with 5-HT a Nernst plot of the basal membrane potential has a slope of 53 mV for a tenfold change in external potassium concentration and the slope at rest deviates from this over the range I-20 mM external potassium. 4. Hyperpolarization of the basal membrane by 5-HT is abolished if the chloride in the bathing medium is replaced by isethionate. 5. The diuretic agent amiloride inhibits fluid secretion by a mechanism which may include a reduction in calcium entry in addition to its recognized effect on sodium permeability. 6. A model is proposed in which fluid secretion is driven by the active transport of potassium across the apical membrane with chloride following passively.

Amiloride

Membrane permeability changes during stimulation of isolated salivary glands of Calliphora by 5-hydroxytryptamine.

1. The membrane resistance of isolated salivary glands was found to decrease in response to 5-HT. The change in resistance was calcium-dependent. 2. The resistance change of the apical membrane was found to be much greater than the change in resistance of the basal membrane. 3. Potential responses under current-clamped conditions showed that one part of the biphasic response to 5-HT (attributed to an increase in chloride permeability) could be reversed and the other part (attributed to an increase in a potassium pump) could not. 4. These observations have been incorporated into a model which, on evaluation, predicts all of the observed potential changes during the action of 5-HT. It suggests that the potential responses reflect changes in the internal chloride concentration produced by the calcium-dependent increases in chloride permeability.

Animals

Calcium content and exchange in frog skeletal muscle.

Calcium content and exchange in frog ELD IV muscle were examined employing the efflux technique. 2. Muscle calcium was found to exchange with four time constants, 21-5 sec 2-7, 32 and 1244 min. 3. All calcium was found to be exchangeable with more than half the total amount residing in an extracellular compartment. 4. Results obtained from ELD IV muscles and single fibres were identical. 5. Muscle calcium content was found to remain constant up to 20 hr in vitro. 6. Extra exchange of calcium occurs upon contraction. This extra exchange appears to occur in the most slowly exchanging component. 7. The data are discussed in relation to morphological and autoradiographic findings and a model of calcium exchange in skeletal muscle is presented.

Animals

Contractile inactivation in frog single denervated muscle fibers.

After preliminary conditioning depolarizations, single muscle fibers of the frog were tested for ability to contract in response to depolarization by 100 mM K+ Ringer solution. Denervated fibers (6-42 days) lose their ability to produce a 100 mM K+ contracture more rapidly than do control fibers. This decrease in 100 mM K+ contracture size (inactivation) is dependent on length of exposure to and magnitude of the conditioning depolarization and on the calcium concentration in the external medium. At 0.4 mM Ca++, the inactivation is 3 times faster than at 1.5 mM Ca++. The rate of contracture loss is not correlated with fiber diameter or the number of days after failure of neuromuscular transmission, and the preliminary conditioning depolarizations do not affect the rate of terminal relaxation from the 100 mM K+ contractures.

Animals

Contractile repriming of single muscle fibers: inactivation and tubular diffusion.

Following maximal potassium contractures of isolated single fibers from Rana pipiens, a brief recovery period is required to restore the ability to produce maximal tension by subsequent depolarization. At normal potassium concentration (2.5 mM), there is a minimal lag time of 5 s before any tension is produced by a second exposure to elevated K+; half-maximal tension can be produced in 13 s. The rate of repriming is directly related to the square of the fiber radius and is slowed by lowering external calcium concentration (1.5-0.4 mM). There is no significant difference in any of these respects between normal fibers and fibers dissected from semitendinosus muscles which had been chronically denervated for periods of up to 8 wk. Increased potassium concentration (5-20 mM) in the recovery solution decreased the steady-state repriming level, affecting denervated fibers more than controls. No repriming occurred at concentrations above 20 mM (-48 mV membrane potential.) At these intermediate concentrations, repriming was less complete in denervated muscle, reflecting the increased speed of delayed contractile inactivation. From the times to 50% repriming, which were proportional to the square of fiber radius, we estimated an upper limit for the diffusion coefficient for K+ in the transverse tubular system of 0.3 X 10(-6) cm2/s for both control and chronically denervated fibers.

Animals

Stimulus-secretion coupling in an insect salivary gland: cell activation by elevated potassium concentrations.

1. Fluid secretion by isolated salivary glands was stimulated by elevating the external potassium concentration. 2. The stimulatory effect of potassium was dependent on external calcium and was potentiated by a subthreshold dose of 5-hydroxytryptamine (5-HT). 3. During the action of 120 mM potassium there was a large calciumdependent decrease in transepithelial resistance similar to that produced with 5-HT at normal potassium concentrations. 4. These results on Calliphora salivary glands are compared with other cases where cells are activated by high potassium. In most cases, the effect of high potassium is dependent upon calcium, suggesting that the latter plays a primary role in cell activation.

5-Hydroxytryptophan

Laser temperature-jump technique for relaxation studies of the ionic conductances in myelinated nerve fibers.

A temperature-jump technique for single nodes of Ranvier has been developed using a pulsed laser system. The temperature perturbation was accomplished by firing the laser beam obtained from a neodymium rod through the solution surrounding a single node. The temperature step was achieved within 1 msec using the laser in the normal mode of operation. During the voltage-clamped steady-state current a temperature jump from 4 degrees C increased the current to a new steady-state value within the time course of the T-jump. This finding suggests that the maximum potassium permeability P(K) has a rapid relaxation time and that the steady-state value of n (the value of potassium permeability divided by its maximum value) is relatively independent of temperature. T-jumps applied during the voltage-clamped sodium currents showed that the sodium permeability changed with a relaxation time that was also shorter than the duration of the normal mode laser output. T-jumps observed during a hyperpolarization or at the resting potential showed no detectable conductance change. When a T-jump immediately preceded a voltage clamp pulse the technique was then used to investigate the effect of changes in the steady-state temperature on the ionic conductances. It was found that the magnitude of the change in membrane current due to a T-clamp was directly related to the level of cathodal polarization.

Animals

Nerve stimulation and electrical properties of frog skin.

The suitability of frog skin glands as a model for the study of secretory mechanisms in exocrine glands was explored. Periodic voltage clamp was used to determine continually the short-circuit current, chord conductance, and electromotive force of frog skin during neural and pharmacological activation of the skin glands. Both the chord conductance and the short-circuit current increased with glandular activation; the temporal dissociation of these increases suggests that there are at least two separate components to the secretory response. The sensitivity of the secretory electrical changes to changes in the ionic composition of the bathing solutions supports the notion of electrogenic chloride active transport as being basic to the activity of the exocrine glands.

Animals