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Biomedical subjects

R D Keynes

Publications and source records attributed to R D Keynes.

At least 37 records · Page 2Linked to original sources
PubMed1984

On the binding of labelled saxitoxin to the squid giant axon.

Measurement of the maximum saturable binding capacity for 3H-labelled saxitoxin in well cleaned giant axons from Loligo forbesi gave a value of 290 +/- 65 (s.e.m.) binding sites per square micrometre of membrane. In samples of the fin nerve the total saturable uptake was 12.4 +/- 2.4 fmol mg-1 wet mass, corresponding to 94 sites per square micrometre for fibres whose mean diameter after fixation and embedding was estimated as 16 micron.

Animals↗
PubMed1984

The effect of local anaesthetics on the components of the asymmetry current in the squid giant axon.

The effects of local anaesthetics and holding potential on sodium and asymmetry currents were studied in intracellularly dialysed squid giant axons. The asymmetry currents were fractionated into their inactivating and non-inactivating components, and the charge displacements Qi and Qn of the two components were determined for pulse potentials between -20 and +40 mV. The charged local anaesthetic RAD 366, a quaternary derivative of lidocaine, applied internally at a concentration in the dialysis solution of 1 mM, did not change Qn, but reduced Qi about 3-fold. The neutral local anaesthetic benzocaine, applied externally at a concentration in the bathing solution of 1 mM, had effects very similar to RAD 366. It did not change Qn, but reduced Qi and the sodium current about 2 . 5-fold. Unlike local anaesthetics, steady membrane depolarization had essentially equal effects on Qn, Qi and sodium current. Lowering the holding potential from -98 to -60 mV for several minutes reduced all three variables to about half. Models of sodium channel voltage-gating are discussed which implicate both Qn and Qi, and which account for the selective blockage of Qi by sodium inactivation and local anaesthetics.

Action Potentials↗
PubMed1983

The Croonian Lecture, 1983. Voltage-gated ion channels in the nerve membrane.

In the Croonian Lecture for 1957, Sir Alan Hodgkin described the role of the channels selective for sodium and potassium ions in the conduction of the nervous impulse. An essential feature of these channels is the manner in which the complex kinetics of their opening and closing is controlled by the electric field across the membrane, and the purpose of the present lecture is to consider the advances that have been made in the past 25 years towards an understanding of the underlying molecular mechanisms. One such advance has been the successful recording, independently of the ionic currents, of the small asymmetry current known as the gating current, that accompanies the conformational changes that take place in the sodium channels. A quantitative analysis of the characteristics of the gating current suggests that activation is brought about by two more or less independent processes operating in parallel, to one of which the slower mechanism of inactivation is coupled sequentially. However, it is clear that a complete picture of the gating system will only be arrived at by combining evidence of this kind with that provided by other new lines of approach such as studies of single ion channels in various tissues by means of fluctuation analysis and the patch-clamping technique, and a reinvestigation of the kinetics of activation of the potassium channels.

Anesthetics, Local↗
PubMed1983

Kinetics of activation of the sodium conductance in the squid giant axon.

The time course of the rise in sodium conductance during positive voltage-clamp pulses was measured in squid giant axons perfused with CsF and immersed in low-sodium solutions. The initial transients were eliminated by subtraction of records made after blocking the sodium channels with tetrodotoxin. The value of tau m as defined by Hodgkin & Huxley (1952) passed through a well defined maximum at a membrane potential of about -35 mV. On fitting the initial inflexion in the rise of INa to the expression mXh instead of m3h, the value of X was found to vary from axon to axon between 2.9 and 4.4, with an average of 3.5. For any given axon, X did not vary significantly with pulse potential. Measurements of tau m were made on approaching each value of the membrane potential both from the negative and from the positive side. The cube law kinetics of the Hodgkin-Huxley equations were closely obeyed. Application of a negative prepulse to -180 mV delayed the rise of conductance by 20 musec at 7 degrees C without obviously changing tau m. Comparisons of the voltage dependence of tau m with that of the time constant tau 1 of the fast relaxation of the asymmetry current measured in the same axon, showed that tau 1 was smaller than tau m except at positive potentials, was less steeply voltage-dependent, and reached its maximum at a more positive potential.

Animals↗
PubMed1982

Fractionation of the asymmetry current in the squid giant axon into inactivating and non-inactivating components.

The operation of the voltage-sensitive sodium gating system in the nerve membrane involves conformational changes that are accompanied by small asymmetrical displacement currents. The asymmetry current may be divided into a component that is inactivated by positive voltage-clamp pulses, and recovers from inactivation with exactly the same time course as the sodium conductance, and one that is not inactivated. A method is described for recording the two components separately with the aid of an inactivating prepulse. They appear to have a marked difference in their rising phases, that of the non-inactivating component being just about as fast as the imposed step in membrane potential, while the inactivating component requires some tens of microseconds to reach its peak.

Animals↗
PubMed1982

The relationship between the inactivating fraction of the asymmetry current and gating of the sodium channel in the squid giant axon.

A quantitative comparison between the voltage dependence of the inactivating component of the asymmetrical charge transfer in the squid giant axon and that of the sodium conductance indicates that activation of the sodium system involves either three subunits operating in parallel or a three-step series mechanism. This is confirmed by an examination of the relative timing of the flow of asymmetry and ionic currents during the opening and closing of the sodium channels. In agreement with previous suggestions, inactivation is coupled sequentially to activation. The evidence appears to argue against a trimeric system with three wholly independent subunits and favours a monomeric system that undergoes a complex sequence of conformational changes.

Animals↗
PubMed1976

Comparative aspects of salt and water transport across lung.

Calculation suggests that the outward flux of water from the surface of the lung into the expired air could be driven by a relatively small standing osmotic gradient. The structure of this alveolar epithelium does not appear to possess the characteristics of a typical secretory epithelium. These considerations argue against the occurrence of active transport in the alveoli of the adult mammalian lung.

Animals↗
PubMed1976

The effect of acetzolamide on Ion transport across isolated sheep rumen epithelium.

1. The net fluxes of sodium and chloride from the lumen to the blood side of isolated sheep rumen epithelium were reduced by treatment of both sides of the epithelium with acetazolamide. 2. The changes in the net fluxes of sodium and chloride were significantly correlated and showed recovery after removal of acetazolamide. 3. The net flux of potassium from blood to lumen side of the epithelim was not affected by treatment with acetazolamide. 4. It is suggested that acetazolamide blocks coupled sodium and chloride transport which may be mediated through 'low-activity' carbonic anhydrase enzymes.

Acetazolamide↗
PubMed1976

The temporal and steady-state relationships between activation of the sodium conductance and movement of the gating particles in the squid giant axon.

1. Comparisons were made between the kinetics and steady-state properties of the sodium conductance changes and of the sodium gating currents, in the squid giant axon perfused with caesium fluoride and maintained at a high membrane holding potential. The conductance measurements were made with reduced external sodium and as much electrical compensation as possible, in order to minimize errors caused by the series resistance. 2. After an initial delay of 10-150 musec whose size was a function of the holding potential and pulse amplitude, the conductance rise on depolarization followed cube law kinetics. 3. Values of the time constant taum, as defined by Hodgkin & Huxley (1952b), were determined for membrane potentials ranging between -140 and +70 mV. They lay on a nearly symmetrical bell-shaped curve with maximum (at 6-3 degrees C) of just under 500 musec at -36 mV. 4. Values of the gating current time constant tau(V) were determined over the same potential range, and found to lie on a very similar bell-shaped curve. A computed least-squares best fit gave the maximum as 460 musec, also falling at about -36 mV. 5. The midpoint of the minfinity curve lay at -34 mV, and its slope at this point was 0-0139 mV-1. Another series of measurements on intact axons gave a midpotential of -25 mV. In the perfused axons the state of the membrane was better described by the constant field equation than by gNa. Recalculation of minfinity from PNa shifted the curve about 15 mV in a positive direction.

Animals↗
PubMed1975

The rate of action of tetrodotoxin on sodium conductance in the squid giant axon.

When tetrodotoxin is applied to or washed away from the squid giant axon, the rates at which the sodium conductatnce is blocked and unblocked are an order of magnitude smaller than those reported for the isolated node of Ranvier. This slowing is to be expected if in squid the tetrodotoxin binding sites act as a saturable sink in series with the barrier to free diffusion imposed by the presence of the Schwann cell. A comparison has been made between the rates observed experimentally and those calculated for a computer model of the system, in order to estimate the apparent density in the membrane of both specific and non-specific tetrodotoxin binding sites. The figure thus obtained for the number of sodium channels in the squid giant axon, several hundred per square micrometre, agrees well with those derived from other lines of argument.

Animals↗