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A L Gorman

Publications and source records attributed to A L Gorman.

At least 37 records · Page 2Linked to original sources
PubMed1981

Time dependence of the calcium-activated potassium current.

We investigated the dependence of the kinetics of the Ca2+-activated K+ current of the molluscan neuron soma upon membrane potential. The K+ current was activated by intracellular Ca2+ ion injection in neurons with blocked inward Na+ and Ca2+ currents. The difference between currents was measured with brief pulses (less than 100 ms) before and immediately after Ca2+ injection and was used as the Ca2+ activated K+ current at difference membrane potentials. The results in normal (10 mM) and in high (200 nM) external K+ show that the time-course of the Ca2+-activated K+ current depends upon membrane voltage and that the current activates more rapidly with membrane depolarization.

Animals↗
PubMed1981

Effects of 4-aminopyridine on potassium currents in a molluscan neuron.

The effects of 4-aminopyridine (4-AP) on the delayed K+ current and on the Ca2+-activated K+ current of the Aplysia pacemaker neurons R-15 and L-6 were studied. The delayed outward K+ current was measured in Ca2+-free artificial seawater (ASW) containing tetrodotoxin (TTX), using brief depolarizing clamp pulses. External (and internal) 4-AP blocks the delayed K+ current in a dose-dependent manner but does not block the leakage current. Our results show that one 4-AP molecule combines with a single receptor site and that the block is voltage dependent with an apparent dissociation constant (K4-AP) of approximately 0.8 mM at 0 mV. K4-AP increases e-fold for a 32-mV change in potential, which is consistent with the block occurring approximately 0.8 of the distance through the membrane electrical field. The 4-AP block appears to depend upon stimulus frequency as well as upon voltage. The greater speed of onset of the block produced by internal 4-AP relative to when it is used externally suggests that 4-AP acts from inside the cell. The Ca2+-activated K+ current was measured in Ca2+-free ASW containing TTX, using internal Ca2+-ion injection to directly activate the K+ conductance. Low external 4-AP concentrations (less than 2 mM) have no effect on the Ca2+-activated K+ current, but concentrations of 5 mM or greater increase the K+ current. Internal 4-AP has the same effect. The opposing effects of 4-AP on the two components of the K+ current can be seen in measurements of the total outward K+ current at different membrane potentials in normal ASW and during the repolarizing phase of the action potential.

4-Aminopyridine↗
PubMed1981

Effects of tetraethylammonium on potassium currents in a molluscan neurons.

The effects of tetraethylammonium (TEA) on the delayed K+ current and on the Ca2+-activated K+ current of the Aplysia pacemaker neurons R-15 and L-6 were studied. The delayed outward K+ current was measured in Ca2+-free ASW containing tetrodotoxin (TTX), using brief depolarizing clamp pulses. External TEA blocks the delayed K+ current reversibly in a dose-dependent manner. The experimental results are well fitted with a Michaelis-Menten expression, assuming a one-to-one reaction between TEA and a receptor site, with an apparent dissociation constant of 6.0 mM. The block depends on membrane voltage and is reduced at positive membrane potentials. The Ca2+-activated K+ current was measured in Ca2+-free artificial seawater (ASW) containing TTX, using internal Ca2+ ion injection to directly activate the K+ conductance. External TEA and a number of other quaternary ammonium ions block the Ca2+-activated K+ current reversibly in a dose-dependent manner. TEA is the most effective blocker, with an apparent dissociation constant, for a one-to-one reaction with a receptor site, of 0.4 mM. The block decreases with depolarization. The Ca2+-activated K+ current was also measured after intracellular iontophoretic TEA injection. Internal TEA blocks the Ca2+-activated K+ current (but the block is only apparent at positive membrane potentials), is increased by depolarization, and is irreversible. The effects of external and internal TEA can be seen in measurements of the total outward K+ current at different membrane potentials in normal ASW.

Animals↗
PubMed1981

Intracellular calcium and the control of neuronal pacemaker activity.

Pacemaker activity of the Aplysia bursting pacemaker neuron R-15 was analyzed. It was shown that the free intracellular Ca2+ concentration, as measured by arsenazo III, increases during the depolarizing phase of the pacemaker cycle and declines throughout the hyperpolarizing phase that follows. This increase in Ca2+ results from the activation of voltage-dependent Ca2+ channels that open during the depolarizing phase of the cycle. The extracellular K+ concentration also increases during the depolarizing phase of the cycle and is correlated with an outward K+ current that opposes the inward current carried by Ca2+ ions. The increase in internal Ca2+ is sufficient to activate a K+ conductance that depends on the magnitude of the change in internal Ca2+ and on membrane potential, which is responsible for the hyperpolarizing phase of the cycle. It is proposed that the membrane oscillation depends on three separate but linked systems, which include a voltage-dependent Ca2+ channel, the internal Ca2+ concentration, and a Ca2+-activated K+ channel.

Action Potentials↗
PubMed1980

Non-uniform Ca2+ buffer distribution in a nerve cell body.

In nerve cells, Ca2+ influx through voltage-dependent channels in the membrane causes a transient rise in the intracellular, free Ca2+ concentration. Such changes have been shown to be important for the release of transmitter at the axon terminal and for the control of the movement of ions through channels in the soma membrane. The transient behaviour of the rise in Ca2+ concentration can, in part, be explained by the presence of sequestering systems in the cell which tend to limit the magnitude and duration of changes in internal Ca2+ (refs 7--10). It is possible that systems involved in buffering changes in internal Ca2+ are not distributed uniformly throughout the cell. This is particularly likely in the cell body, where a significant portion of the cytoplasm is occupied by the nucleus, whose buffering capacity may differ from that of other cellular regions. We report here that in the soma of a molluscan pacemaker neurone, the machinery responsible for short-term buffering of Ca2+ ions is localized near the inner surface of the plasma membrane.

Animals↗
PubMed1980

Intracellular calcium accumulation during depolarization in a molluscan neurone.

1. The bursting pacemaker neurone R-15 of Aplysia was injected with the Ca2+ sensitive dye arsenzo III. Changes in absorbance were measured with a differential spectrophotometer to monitor changes in free intracellular Ca2+ during membrane depolarization under voltage clamp conditions. 2. Dye absorbance increased linearly for depolarizing pulse durations up to 100 msec and approximately linearly between 100 and 300 msec, but for longer durations the absorbance change decreased. 3. The absorbance change vs. voltage relation increased steeply between -20 and 0 mV (e-fold per 8.5 mV), peaked at +36 mV and declined non-linearly to an estimated null or suppression potential of about +139 mV. 4. TTX (5 x 10(-5 M) had no effect on the change in dye absorbance produced by brief or long duration stimuli whereas Ca2+ free ASW abolished all changes in dye absorbance. 5. The absorbance change saturated with increasing external Ca2+ concentrations. The relation between dye absorbance and external Ca2+ concentration was hyperbolic and for a small range of external Ca2+ concentration and membrane potentials could be fitted by a Michaelis--Menten expression where the dissociation constant and the maximum absorbance change are voltage dependent. 6. The absorbance change was reduced by external divalent ions which block the Ca2+ channel (e.g. Cd2+ and Ni2+). The suppression of dye absorbance was increased by membrane depolarization and suggests that there is a voltage dependent site within the Ca2+ channel which binds divalent ions. 7. The decline of the absorbance--voltage relation from its peak to the suppression potential showed a greater nonlinearity when longer duration voltage clamp pulses were used. The non-linearity can be explained if the accumulation of Ca2+ ions next to the inner surface of the membrane during depolarization reduces the driving force on Ca2+ ions and thus decreases Ca2+ ion influx. 8. The suppression potential estimated from the absorbance--voltage relation increased 29 mV per tenfold change in the external Ca2+ concentration and thus can be used to estimate the Ca2+ equilibrium potential. 9. The change in dye absorbance produced by brief depolarizing voltage clamp steps was inactivated at positive holding potentials (50% inactivation at about -14 mV). Our results suggest that the slow decrease in dye absorbance during prolonged depolarization is caused by inactivation of the Ca2+ channel.

Animals↗
PubMed1980

Potassium conductance and internal calcium accumulation in a molluscan neurone.

1. The Aplysia neurone R-15 was injected with the Ca(2+) sensitive dye arsenazo III. Changes in dye absorbance were measured with a differential spectrophotometer to monitor changes in the free internal Ca(2+) concentration, [Ca](i), during membrane depolarization and during intracellular Ca(2+) ion injection under voltage clamp conditions.2. The absorbance change, and thus [Ca](i), increases linearly with Ca(2+) injection intensity at constant duration. The absorbance change produced by a constant intensity Ca(2+) injection also increases with injection duration, but this increase is asymptotic.3. The Ca(2+) activated K(+) current, I(K, Ca), increases linearly with the increase in [Ca](i) and its rise and decay follows closely the time course of the absorbance change produced by internal Ca(2+) injection.4. The Ca(2+) activated K(+) conductance increases exponentially with membrane depolarization. The increase in K(+) conductance activated by a constant intensity and duration Ca(2+) injection is on average e-fold for a 25.3 mV change in membrane potential.5. The difference in net outward K(+) current measured during depolarizing pulses to different membrane potentials in normal and in Ca(2+) free ASW was used as an index of I(K, Ca). Its time course was approximately linear for the first 50-100 msec of depolarization, but for longer times the relation approached a maximum. Simultaneous measurements of the arsenazo III absorbance changes were broadly consistent with the activation of I(K, Ca) being brought about by the rise in [Ca](i) during a pulse.6. The relation between Ca(2+) activated K(+) conductance and membrane potential is bell shaped and resembles the absorbance vs. potential curve, but its maximum is displaced to more positive membrane potentials. The shift in the two curves on the voltage axis can be explained by the potential dependence of G(K, Ca).7. The net outward K(+) current measured with depolarizing voltage pulses in normal and in Ca(2+) free ASW is increased when [Ca](i) is elevated by internal Ca(2+) injection. With large and prolonged Ca(2+) injections the net outward current is depressed following the decline of [Ca](i).8. The time and frequency dependent depression of the net outward K(+) current which occurs during repetitive stimulation is shown to have no obvious temporal relation to the increase in [Ca](i). The depression is relieved by an increase in [Ca](i) caused by internal Ca(2+) injection.9. The net outward K(+) current measured with brief depolarizing pulses which approach the estimated Ca(2+) equilibrium potential and therefore do not cause Ca(2+) influx and accumulation is facilitated by a previous depolarizing pulse which causes a rise in [Ca](i)..10. The facilitation experiments also suggest that the activation of I(K, Ca) by [Ca](i) has a significant time constant. During a depolarizing pulse, the rise in [Ca](i) next to the membrane, and hence I(K, Ca) is expected to follow the square root of time, but a delay in the activation of I(K, Ca) by [Ca](i) could explain why the observed time course of I(K, Ca) is initially almost linear.11. The potential dependence of the Ca(2+) activated K(+) conductance can be explained if the internal Ca(2+) binding site is about half way through the membrane.

Animals↗
PubMed1979

External and internal effects of tetraethylammonium on voltage-dependent and Ca-dependent K+ currents components in molluscan pacemaker neurons.

Ca-dependent and voltage-dependent K+ currents in Aplysia pacemaker neurons were studied separately by electrophoretic injections of Ca2+ ions into the cells to induce the Ca-dependent component and by step depolarizations in the absence of external Ca2+ ions to induce the voltage-dependent component. Intracellular tetraethylammonium (TEA) blocks the voltage-dependent but not the Ca-dependent K+ current. Extracellular TEA blocks both components reversibly but is more effective in inhibiting the Ca-dependent K+ currents. Our results suggest that there are TEA receptor sites for the voltage-dependent K+ current on each side of the membrane, but only an external receptor site for the Ca-dependent K+ current.

Animals↗
PubMed1979

Contribution of calcium and potassium permeability changes to the off response of scallop hyperpolarizing photoreceptors.

1. The membrane response of the distal photoreceptors in the retina of the scallop Pectin irradians to the termination of a bright white light (off response) is shown to be composed of the decay of the hyperpolarizing receptor potential and an action potential with slow kinetics. 2. The action potential can be produced in darkness in the absence of external Na+ ions by membrane depolarization. 3. The action potential is maintained by replacement of external Ca2+ with Sr2+ or Ba2+, but not by Mg2+. In normal external Ca2+ (9mM), the action potential is abolished by the addition of the Ca2+ inhibitors, La3+, Co2+, and Mn2+ or the organic Ca2+ antagonist D-600. 4. Elevated external Ca2+ concentrations increase the rate of rise and peak amplitude of the action potential as well as the rate of repolarization and after hyperpolarization, but decrease the duration. 5. The rate of rise and peak amplitude of the action potential are increased by the K+ antagonists tetraethylammonium (TEA) 4-amino-phyridine (4-AP), Ba2+ and procaine. The antagonists have different effects on subsequent phases of the response, however. External TEA and Ba2+ increase the duration, but decrease the rate of repolarization and abolish the after hyperpolarization, whereas external 4-AP and procaine increase the rate of repolarization, decrease the duration and increase the after hyperpolarization. 6. The ratio of the Ca2+ to K+ permeability (P Ca/P K) estimated from the constant field equation at the peak of the action potential in different external Ca2+ concentrations is close to 1. 7. The maximum rate of rise and the peak amplitude of the action potential are increased by membrane hyperpolarization and decreased by membrane depolarization. They are decreased by background light intensity relative to their value in the dark. 8. In normal ASW the action potential can be identified during the off response as a small overshoot of membrane potential relative to its value in the dark. 9. The rate of repolarization of the off response in normal ASW is reduced by agents or conditions which inhibit or reduce Ca2+ permeability changes, e.g. external Co2+ and La2+ or zero external Ca2+. 10. Our results suggest that a voltage-dependent increase in membrane permeability to Ca2+ and to K+ ions modifies the repolarizing phase of the receptor potential.

Action Potentials↗
PubMed1979

Internal effects of divalent cations on potassium permeability in molluscan neurones.

1. Electrophoretic injection of Ca ions into Aplysia pace-maker neurones activates an outward current, carried primarily by K ions, whose magnitude is determined by the intensity and duration of the injection current, the position of the injection electrode within the cell and the holding potential. 2. The efflux of K ions measured with an extracellular K sensitive electrode is a linear function of the Ca activated outward current and disappears at its reversal potential. 3. The outward current decays exponentially with an early and late phase. The early but not the late phase is temperature dependent with a Q10 of about 3-5. 4. Of the divalent cations which activate the outward current, Ca is the most effective followed by Cd, Hg, Sr, Mn and Fe. Injections of Ba, Co, Cu, Mg, Ni and Zn are ineffective. 5. Low temperatures or prolonged injection of Cd or Hg, increase the amplitude of the outward current activated by Ca. 6. Prolonged injection of Ba inhibits the Ca activated outward current and reduces substantially all currents carried by K ions. 7. It is concluded that the effectiveness of a divalent cation in activating the K current is, in part, related to its ionic radius, and that the site of activation can accommodate ionic radii between about 0.76 and 1.13 A.

Animals↗
PubMed1978

Ionic effects on the membrane potential of hyperpolarizing photoreceptors in scallop retina.

1. The effects of different external ionic conditions and of metabolic inhibitors on the membrane potential of hyperpolarizing photoreceptors in the retina of the scallop Pecten irradians were examined in the presence and absence of light.2. Changes in extracellular K(+) have a greater effect on membrane potential in the light than in darkness. The receptor potential is increased in amplitude when [K](o) is reduced and decreased when [K](o) is elevated. It is hyperpolarizing when [K](o) is less than the estimated value for [K](i) and depolarizing when this condition is reversed.3. The complete replacement of [Na](o) causes a significant hyperpolarization of membrane potential in darkness, whereas it has a much smaller hyperpolarizing effect on the peak of the receptor potential.4. The ratio of Na(+) to K(+) permeabilities (P(Na)/P(K)) decreases during bright illumination. Our results suggest that P(K) is seven times that for P(Na) in the dark but is 57 times greater than P(Na) in light.5. The metabolic inhibitors DNP and NaCN cause membrane potential in the dark to hyperpolarize. This hyperpolarization is associated with a decrease in the P(Na)/P(K) ratio similar to that found during illumination.6. High [Ca(+)](o) also causes membrane potential in the dark to hyperpolarize. This hyperpolarization is associated with an increase in membrane conductance.7. The results indicate that the hyperpolarizing receptor potential of the distal photoreceptor is produced by a light-evoked increase in K(+) permeability.

Animals↗
PubMed1978

Changes in the intracellular concentration of free calcium ions in a pace-maker neurone, measured with the metallochromic indicator dye arsenazo III.

1. The bursting pace-maker R-15 cell of Aplysia was injected with the Ca2+ sensitive dye arsenazo III. Changes in absorbance were measured with a differential spectrophotometer to monitor changes in free intracellular Ca2+, [Ca-a], during activity. 2. Dye absorbance increased during each pace-maker-induced burst of action potentials and decreased during the hyperpolarizing phase of the pace-maker cycle. 3. The increase in dye absorbance was, at least in part, dependent upon action potential discharge and was greater when action potential duration was prolonged by treatment with tetraethylammonium chloride. 4. Changes in dye absorbance occurred under voltage clamp conditions when the membrance was depolarized 5-15 mV from a holding potential near the resting potential and were larger with greater step depolarizations. 5. These changes were completely blocked by the addition of 3mM-La3+ to normal ASW. 6. The ratio of the absorbance change between two pairs of wave-lengths during the pace-maker cycle was compared with the ratio observed following injection of Ca2+, Mg2+ and H+ ions. The ratio for the pace-maker cycle was well matched by that for Ca2+ injection, but not by that for injection of Mg2+ or H+. 7. Intracellular Ca2+ injections which increased [Ca]1 to the same amount as occurred during the pace-maker cycle also produced an outward current of sufficient magnitude to account for the post-burst hyperpolarization. 8. Depolarization of the cell membrane by extrinsic current during the burst increased and prolonged the change in dye absorbance as well as the post-burst hyperpolarization. 9 It is suggested that Ca2+ enters during the pace-maker cycle, thereby increasing [Ca]i, and that this increase is sufficient to activate an outward current carried by K+ ions which causes or contributes to the post-burst hyperpolarization.

Action Potentials↗
PubMed1977

Internal calcium changes in a bursting pacemaker neuron measured with arsenazo III.

Arsenazo III was used to measure changes in the free intracellular calcium concentration during spontaneous bursting pacemaker activity in the Aplysia R1K neuron. Intracellular calcium increased during the burst, and this increase was sufficient to cause the hyperpolarization that followed. The results suggest that the interval between bursts is determined by the rate of subsequent decline of free intracellular calcium.

Action Potentials↗
PubMed1974

Control of membrane N+ permeability in a hyperpolarizing photoreceptor: similar effect of light and metabolic inhibitors.

In the hyperpolarizing photoreceptors of the scallop Pecten irradians the metabolic inhibitors cyani 'e and 2,4-dinitrophenol cause a rapid hyperpolarization and increase in membrane permeability to potassium ions, similar to the effect of light. Cellular metabolism appears important in maintaining the low permeability to potassium ions necessary to keep the membrane depolarized in darkness, possibly by regulating the intracellular calcium ion concentration.

Animals↗
PubMed1974

Steady-state contribution of the sodium pump to the resting potential of a molluscan neurone.

1. The electrogenic contribution of the Na(+)-K(+) exchange pump to the membrane potential of the Anisodoris giant neurone (G cell) was examined under steady-state and Na(+) loaded conditions.2. The membrane potential was variable for the first 1-4 hr after impalement, but, in the absence of experimental manipulation, remained constant thereafter. The average membrane potential for ten cells maintained at 11-13 degrees C and measured 5-36 hr after impalement was 55.8 +/- 1.0 mV (S.E. of mean).3. Low concentrations of external ACh caused a reversible increase in membrane Na(+) conductance. Brief exposure to ACh proved a fast and reversible technique to load the cell with Na(+) ions, and transiently stimulate the electrogenic Na(+) pump.4. In ten cells maintained from 5 to 36 hr at 11-13 degrees C the reduction in membrane potential produced by inhibition of the Na(+) pump with ouabain was remarkably constant between cells and averaged + 9.7 mV.5. Cells maintained under steady-state conditions (at 11-13 degrees C) for extended periods of time were shown to be relatively insensitive to changes in temperature and to small changes in external K(+).6. It is estimated that the Na(+)-K(+) exchange pump contributes approximately - 10 mV to the steady-state resting potential of the G cell, and that two Na(+) ions are extruded for every K(+) ion transported into the cell per pump cycle.

Acetylcholine↗