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Biomedical subjects

E Rojas

Publications and source records attributed to E Rojas.

At least 253 records · Page 14Linked to original sources
PubMed1971

The binding of tetrodotoxin to nerve membranes.

1. The% reduction in size of the externally recorded action potential produced by concentrations of tetrodotoxin (TTX) in the range 6-300 nM was determined for the small non-myelinated fibres of the rabbit cervical vagus nerve and of the walking leg nerves of crab and lobster. The concentration of TTX for 50% reduction was around 80 nM for rabbit vagus and 14 nM for crab nerve.2. Bio-assay procedures were devised to measure the amount of TTX taken up by a nerve when it was exposed to a very small volume of a solution whose TTX content was just great enough to produce 100% block of conduction. The extracellular space of each nerve was determined with [(14)C]sugar so that an allowance could be made for extracellular dilution.3. The TTX binding by rabbit, crab and lobster nerve was respectively 0.064, 0.053 and 0.036 p-mole/mg wet weight of nerve.4. The binding of saxitoxin was measured in rabbit vagus nerve, and found to be much the same as that of TTX.5. Control experiments on rabbit sciatic nerve, where the area of excitable membrane was much smaller, showed that there was relatively little unspecific binding of TTX.6. In view of the evidence presented here and elsewhere that the blocking of sodium conductance by TTX involves the attachment of only one TTX molecule at each sodium site, and that unspecific binding of TTX does not cause serious errors, these results suggest that in 1 mum(2) of nerve membrane the number of sodium sites is 75 for rabbit, 49 for crab, and 36 for lobster nerve.

Action Potentials↗
PubMed1971

Analysis of the potential-dependent changes in optical retardation in the squid giant axon.

1. An analysis has been made of the change in optical retradation of the membrane elicited by the application of voltage-clamp pulses in squid giant axons.2. The retardation response consists of three separate voltage-dependent components. For freshly mounted axons, defined as being in state 1, hyperpolarizing pulses give a rapid increase in the light intensity measured with crossed polarizers which has been termed the fast phase. This is followed by a rather slow return towards the base line termed the rebound. On treatment of the axon with certain agents that include tetrodotoxin, high calcium and terbium, the rebound disappears and the fast phase slows down, increases in size, and has a new slow component added to it. This transition from state 1 to a second state, 2, appears to be irreversible.3. In state 1, the time constant of the fast phase is 20-40 musec at 13 degrees C; it has a very large negative temperature coefficient (Q(10) = Ca.(1/8)). The size of the retardation change is independent of temperature and varies as the square of the applied voltage, but the voltage-retardation curve is symmetrical about a point well beyond zero membrane potential, at an internal potential of around + 70 mV. In state 2, the time constant is about five times larger, and varies much less markedly with temperature; the apex of the voltage-retardation curve is shifted to + 200 mV.4. The rebound has a time constant of the order of 20 msec at 13 degrees C. A 10 degrees rise in temperature more than halves the time constant and roughly doubles the amplitude of the rebound. The voltage dependence of the rebound differed from that of the fast phase.5. The slow component of state 2 has a time constant of about 2 msec which does not change noticeably between 10 and 25 degrees C. The size of this component seems to be linearly dependent on the applied voltage, rather than obeying a square law.6. A tenfold increase in external calcium concentration had no discernible effect on the fast and slow phases, but reversibly reduced the amplitude of the rebound nearly to half.7. In experiments on perfused axons, the retardation response was not measurably altered by any of the modifications made to the composition of the perfusing fluid.8. There was some indication of the possible existence of a small current- or conductance-dependent component of the retardation response.9. These phenomena seem likely to originate either from molecular relaxation processes analogous with the Kerr effect, or from changes in membrane thickness under the influence of the pressure exerted by the electric field. However, the specific molecules involved in the retardation response cannot yet be identified.

Animals↗
PubMed1970

Time course of the sodium influx in squid giant axon during a single voltage clamp pulse.

1. Sodium influx measurements were carried out on internally perfused squid giant axons under conditions of membrane potential control.2. The ratio ;measured extra sodium influx/calculated ionic influx from the inward current record' is close to unity and is independent of the duration of the rectangular pulse of membrane potential.3. The variation of the sodium permeability with time during the voltage clamp pulse is obtained by subtracting the sodium influx obtained from the ;tail' current record from the measured extra sodium influx.4. Evidence was obtained indicating that the measured change in sodium permeability during a sudden decrease of the membrane potential from its resting level involves two processes, namely, a transient increase in sodium permeability (activation), and a decrease in sodium permeability (inactivation).5. The extra influx of sodium during a voltage clamp pulse is not decreased by raising the internal sodium concentration from 0 to 100 mM, supporting the validity of the independence principle.

Animals↗
PubMed1970

Sodium and potassium conductance changes during a membrane action potential.

1. A method for turning a membrane potential control system on and off in less than 10 musec is described. This method was used to record membrane currents in perfused giant axons from Dosidicus gigas and Loligo forbesi after turning on the voltage clamp system at various times during the course of a membrane action potential.2. The membrane current measured just after the capacity charging transient was found to have an almost linear relation to the controlled membrane potential.3. The total membrane conductance taken from these current-voltage curves was found to have a time course during the action potential similar to that found by Cole & Curtis (1939).4. The instantaneous current voltage curves were linear enough to make it possible to obtain a good estimate of the individual sodium and potassium channel conductances, either algebraically or by clamping to the sodium, or potassium, reversal potentials. Good general agreement was obtained with the predictions of the Hodgkin-Huxley equations.5. We consider these results to constitute the first direct experimental demonstration of the conductance changes to sodium and potassium during the course of an action potential.

Action Potentials↗
PubMed1970

Time course of the sodium permeability change during a single membrane action potential.

1. A method for measuring the time course of ionic fluxes during a non-propagated membrane action potential is described; the technique combines intracellular perfusion of the squid giant axon with radioactive tracer methods and with a method for controlling the membrane potential during small time intervals.2. The method is used to determine the temporal course of the sodium extra influx during an action potential.3. The results agree with the time course of the permeability change to sodium ions calculated with the Hodgkin & Huxley equations.4. An average extra sodium influx of 7.13 p-mole/cm(2) per action potential was determined at 12 degrees C and of 5.23 p-mole/cm(2) per action potential at 15 degrees C.

Action Potentials↗
PubMed1969

Analysis of the effects of calcium or magnesium on voltage-clamp currents in perfused squid axons bathed in solutions of high potassium.

Isolated axons from the squid, Dosidicus gigas, were internally perfused with potassium fluoride solutions. Membrane currents were measured following step changes of membrane potential in a voltage-clamp arrangement with external isosmotic solution changes in the order: potassium-free artificial seawater; potassium chloride; potassium chloride containing 10, 25, 40 or 50, mM calcium or magnesium; and potassium-free artificial seawater. The following results suggest that the currents measured under voltage clamp with potassium outside and inside can be separated into two components and that one of them, the predominant one, is carried through the potassium system. (a) Outward currents in isosmotic potassium were strongly and reversibly reduced by tetraethylammonium chloride. (b) Without calcium or magnesium a progressive increase in the nontime-dependent component of the currents (leakage) occurred. (c) The restoration of calcium or magnesium within 15-30 min decreases this leakage. (d) With 50 mM divalent ions the steady-state current-voltage curve was nonlinear with negative resistance as observed in intact axons in isosmotic potassium. (e) The time-dependent components of the membrane currents were not clearly affected by calcium or magnesium. These results show a strong dependence of the leakage currents on external calcium or magnesium concentration but provide no support for the involvement of calcium or magnesium in the kinetics of the potassium system.

Animals↗
PubMed1969

Sodium influxes in internally perfused squid giant axon during voltage clamp.

1. An experimental method for measuring ionic influxes during voltage clamp in the giant axon of Dosidicus is described; the technique combines intracellular perfusion with a method for controlling membrane potential.2. Sodium influx determinations were carried out while applying rectangular pulses of membrane depolarization. The ratio ;measured sodium influx/computed ionic flux during the early current' is 0.92 +/- 0.12.3. Plots of measured sodium influx and computed ionic flux during the early current against membrane potential are very similar. There was evidence that the membrane potential at which the sodium influx vanishes is the potential at which the early current reverses.

Animals↗
PubMed1968

Sodium movements in perfused squid giant axons. Passive fluxes.

Sodium movements in internally perfused giant axons from the squid Dosidicus gigas were studied with varying internal sodium concentrations and with fluoride as the internal anion. It was found that as the internal concentration of sodium was increased from 2 to 200 mM the resting sodium efflux increased from 0.09 to 34.0 pmoles/cm(2)sec and the average resting sodium influx increased from 42.9 to 64.5 pmoles/cm(2)sec but this last change was not statistically significant. When perfusing with a mixture of 500 mM K glutamate and 100 mM Na glutamate the resting efflux was 10 +/- 3 pmoles/cm(2)sec and 41 +/- 10 pmoles/cm(2)sec for sodium influx. Increasing the internal sodium concentration also increased both the extra influx and the extra efflux of sodium due to impulse propagation. At any given internal sodium concentration the net extra influx was about 5 pmoles/cm(2)impulse. This finding supports the notion that the inward current generated in a propagated action potential can be completely accounted for by movements of sodium.

Action Potentials↗