Axonal Membrane Potential Research Articles

Effects of the α-cyano group in the benzyl alcohol moiety on insecticidal and neurophysiological activities of pyrethroid esters

This work was undertaken to clarify the criteria for classifying pyrethroids into Types I and II on the basis of their structure-activity pattern. Thus, effects on the membrane potential in crayfish giant axons of substituted benzyl esters of chrysanthemic, α-substituted iso-valeric, and related carboxylic acids with and without the cyano group at the α-position of the alcohol moiety were examined by an intracellular recording technique. Some α-cyano-benzyl alcohol esters caused a remarkable depolarizing afterpotential in response to a single stimulus similar to that induced by esters without the cyano group. Their effects on sodium currents were examined with an internally perfused crayfish axon under voltage clamp conditions using the sucrose gap method. Irrespective of the presence of the α-cyano group in the alcohol moiety, these compounds induced a tail current upon a step repolarization of the membrane, although the α-cyano compounds decreased the rate of fall of the tail current much more slowly than that of the corresponding compounds without the cyano group. Insecticidal activity of compounds was measured against the American cockroaches under synergistic conditions with metabolic inhibitors. The activity of compounds with the α-cyano group was much higher than that of the corresponding compounds without the cyano group. The higher the tail current activity, the higher was the insecticidal activity, although contributions of such structural effects as hydrophobicity may differ between the two activities. The results suggest that the presence of the cyano group at the α-position of the benzyl alcohol moiety of synthetic pyrethroids enhances both the sodium current and insecticidal activities but is by no means a sufficient structural requirement to differentiate the types of each of the biological activities.

Optical recording of electrical activity from axons and glia of frog optic nerve: potentiometric dye responses and morphometrics.

Voltage-sensitive dyes were used to study the changes in membrane potential in axons and glial cells of the frog optic nerve following electrical stimulation. The lack of a signal in the unstained nerve and the multiphasic action spectra after staining indicated that the optical responses were from the extrinsic dyes. Changes in dye absorption and fluorescence had rapid and slow phases. The rapid phases resulted from action potentials in myelinated and unmyelinated axons. The kinetics of the slow phase of the optical response were similar to the depolarization recorded from the glial cells with intracellular electrodes. The ratio of the amplitudes of the fast and slow phases was characteristic for each type of dye. Pharmacological analysis of the action potential of the unmyelinated axons revealed tetrodotoxin-sensitive sodium channels and 4-aminopyridine-sensitive potassium channels. Repeated exposure of the stained preparation to light led to photodynamic damage as shown by a block of recovery of the glial depolarization. An electron microscopic morphometric study of the nerve was carried out in an effort to understand the contribution of the various anatomical elements to the compound optical response. The ratio of unmyelinated axon membrane to glial membrane was much greater than was the ratio of the fast and slow components of the signal, suggesting that the dyes either had a higher affinity for glial membrane or did not penetrate the nerve uniformly.

Voltage-clamp analysis of a crayfish rectifying synapse.

1. The rectifying crayfish giant motor synapse has been studied in the second abdominal ganglion, using the double-voltage-clamp technique which allowed direct measurements of junctional current at various fixed transjunctional potentials. 2. The transjunctional potential (Vj), defined as the difference between the voltages recorded in the lateral giant axon and the giant motor fibre, was varied from -70 to +50 mV, the minimum and maximum junctional chord conductances (gmin and gmax, respectively) were found to be 1.2 +/- 1.3 microS (n = 10) and 22.9 +/- 6.3 microS (n = 10), respectively. 3. For a given Vj, changes in the lateral giant axon or giant motor fibre membrane potential over a range of +/- 30 mV around their resting levels did not influence the junctional permeability (gj), indicating that the inside-outside potential of the junctional channel does not control gj. 4. Therefore, the steady-state junctional chord conductances were dependent only upon Vj. 5. The voltage dependence of the chord conductance was well fitted by a modified Boltzmann relation given by the equation (Formula: see text) with the constants: A = 0.15 +/- 0.03 mV-1 (n = 10) and V0 = 28 +/- 4 mV (n = 10); the latter two parameters were also found to be independent of both transmembrane potentials. 6. The junctional currents were already constant 1 ms after step changes in the junctional voltage; this was three orders of magnitude faster than the other known examples of voltage-controlled gap junctions between embryonic cells. 7. Our results may be interpreted by a highly voltage-dependent probability of opening of the junctional channels. They also suggest that the gap-junction channels forming the giant motor synapse respond very rapidly to potential and that the hemi-channels which constitute them may not be symmetric.

Axonal contribution to subthreshold currents inaplysia bursting pacemaker neurons

The contribution of axonal activity to the ionic currents which generate bursting pacemaker activity was studied by using the two-electrode voltage-clamp technique in Aplysia bursting neuron somata in conjunction with intraaxonal voltage recordings. Depolarizing voltage-clamp pulses applied to bursting cell somata triggered axonal action potentials. The voltage-clamp current recording exhibited transient inward current "notches" corresponding to each of the axonal spikes. The addition of 50 microM tetrodotoxin (TTX) to the bathing medium blocked the fast axonal spikes and current notches, revealing a slower axonal spike which was blocked by the replacement of external Ca2+ with Co2+. The inward current evoked by applying a depolarizing voltage-clamp pulse in the soma is distorted by the occurrence of the axonal Ca2+ spike. Elimination of the axonal spike, by injecting hyperpolarizing current into the axon, changes both the time course and the magnitude of the inward current. The axonal Ca2+ spikes are followed by a series of Ca2+-dependent afterpotentials: a rapid postspike hyperpolarization, a depolarizing afterpotential (DAP) and, finally, a long-lasting postburst hyperpolarization. The long-lasting hyperpolarization is not blocked by 50 mM external tetraethyl ammonium, an effective blocker of Ca2+-activated K+ current [IK(Ca)], and does not appear to reverse at EK. Hence, the axonal long-lasting hyperpolarization may not be due to IK(Ca). Somatic voltage-clamp pulses in bursting neurons are followed by a slow inward tail current, which is sometimes coincident with a DAP in the axon. In some cells, the amplitude of the slow inward tail current is greatly reduced if axonal spikes and DAPs are prevented by hyperpolarization of the axon, while, in other cells, elimination of axonal activity has little effect. Therefore, the slow inward tail current is not necessarily an artifact of poor voltage-clamp control over the axonal membrane potential but probably results from the activation of an ionic conductance mechanism located partly in the axon and partly in the soma.

Structures of attractors in periodically forced neural oscillators

Periodically forced oscillations in the membrane potential of squid giant axons are analysed by stroboscopic plots at multi-phases of the forcing function. The analysis clarifies the global structures of the attractors in the phase space R 2 × S 1.

Voltage dependence of the Na/Ca exchange in voltage-clamped, dialyzed squid axons. Na-dependent Ca efflux.

A combination of the voltage-clamp and the intracellular dialysis techniques has been used to study the membrane potential dependence of the Nao-dependent Ca efflux in squid giant axons. In order to improve axon survival, experiments were carried out using internal solutions prepared with large impermeant organic anions and cations, which did not affect the operation of the Na/Ca exchange mechanism. In axons dialyzed with solutions prepared without internal Na, the Nao-dependent Ca efflux had a small sensitivity to membrane potential changes. For a 25-mV membrane displacement in the hyperpolarizing direction, the basal Ca efflux increased by only 7.4% (n = 13). When the dialysis medium contained Na (from 20 to 55 mM), the efflux increased 32.3% (n = 25) for the same membrane potential change. The K1/2 for this effect is approximately 5 mM Na, and saturation appears to occur at a Na concentration above 20 mM. Adding ATP to the dialysis medium increased the magnitude of the Nao-dependent Ca efflux without changing its voltage sensitivity. Wide changes in the intracellular ionized Ca concentration (from 0.1 to 230 microM) did not modify the voltage sensitivity of the exchange system. Elimination of the reversal of Na/Ca exchange (Nai-dependent Ca influx) by removing Cao did not modify the voltage sensitivity of the Nao-dependent Ca efflux. When the axon membrane potential was submitted to prolonged changes, the corresponding changes in the Ca efflux were not sustained, but declined exponentially to intermediate values. This effect may indicate a slow inactivation process in the Na/Ca exchange mechanism. Voltage-clamp pulse experiments revealed: (a) the absence of a fast inactivation process in the Na/Ca exchange, and (b) that the activation of the carrier for hyperpolarizing pulses occurs as rapidly as 1 ms.

The kinetics of recovery and development of potassium channel inactivation in perfused squid (Loligo pealei) giant axons.

K+ currents were studied at a normal (-69 mV) and at a depolarized (-49 mV) membrane potential in voltage-clamped squid giant axons perfused with 350 mM-K+ and bathed in K+-free artificial sea water containing tetrodotoxin to block the Na+ channels. Steady-state and instantaneous K+ currents were reduced by over 50% at corresponding voltages at the depolarized membrane potential. Instantaneous chord conductance-voltage curves showed that the depolarized membrane potential caused a uniform reduction of K+ conductance across the voltage range under study. The driving force for K+ ions was comparable at both membrane potentials when a short (2 ms) pre-pulse was used to open the K+ channels. When a longer (7.5 ms) pre-pulse was used, the driving force was actually larger at the depolarized membrane potential. The depolarized membrane potential did drive some K+ ions into the periaxonal space. The amount of K+ ions driven into the periaxonal space was estimated by two independent methods, with similar results. The resulting increase of K+ ions in the periaxonal space (10 mM) was about 40 times too small to account for the large reduction in currents in terms of a reduced driving force for K+ ions. The kinetics of recovery and development of inactivation were monitored by repeatedly applying a 7.5 ms test pulse followed by a long conditioning potential. Both recovery and development of inactivation, from the depolarized membrane potential, were described by the sum of two exponential terms plus a constant. The time constant-voltage curves for both phases of inactivation peaked at about -54 mV at 10 degrees C. The time constant of the slow phase of inactivation at -54 mV was about 12.4 s, while the corresponding time constant for the fast phase was about 2.3 s. The slow relaxation had an apparent plateau of about 11 s at more depolarized membrane potentials. Recovery from inactivation was rapid at hyperpolarized membrane potentials. The steady-state inactivation curve of the K+ channel was incomplete in the depolarizing region; and apparent plateau was reached with about 75% of the K+ current inactivated. The temperature sensitivity of both phases of inactivation corresponded to a Q10 of about 3. Elevated external concentrations of K+ ions did not block either phase of the inactivation process, although the kinetics of recovery from inactivation were slightly faster under these conditions.(ABSTRACT TRUNCATED AT 400 WORDS)

The nature of the membrane potential of glial cells associated with the medial giant axon of the crayfish

Electrophysiological, pharmacological and electron microscopic methods were used to characterize the satellite glial (Schwann-like) cells associated with the medial giant axon of the crayfish, Procambarus clarkii. The satellite glial cell layer surrounding the axon is formed by 15–20 cells deeply interdigitated into each other, forming a vast system of intercellular channels which communicate the axo-glial space with the external medium. The satellite cell layer varies from 0.2–3 μm in thickness. Membrane potentials of the giant axon and the satellite glial cells were monitored before, during and after treatment with ouabain and a variety of cholinergic agonists and antagonists. The membrane potentials of 63 control satellite cells averaged−42.6 ± 0.6 mV. The intracellular localization of the glial cell potential difference was corroborated by lithium carminate marking of the microelectrode tip recording site. Superfusion of satellite cells with 10 −7 m carbachol, nicotine or acetylcholine caused a 15 to 20 mV hyperpolarization from resting level. Muscarine (10 −6 m) had no effect on the glial cell potential. The effect of nicotine was prevented or reversed by-tubocurarine (10 −9 m). The effects of cholinomimetics were reversed by washing the cells in drug-free solution. None of these agents had an obvious effect on axon membrane potential or action potential generation at the concentration used in this study. Ouabain (10 −3 m) also caused a rapid hyperpolarization of the glial cells. The effect lasted 15–18 min after which the membrane rapidly depolarized. The results suggest that (1) the satellite glial cell of the crayfish giant axon system may be studied simultaneously with the axon using electrophysiological techniques; (2) the satellite glial cell membrane appears to have typical acetylcholine receptors of the nicotinic type; (3) the membrane potential of the glial cell is sensitive to ouabain and (4) properties of the glial cell associated with the giant axon of the crayfish are similar to those of the Schwann cell of the tropical squid Sepioteuthis sepioidea.

Modulation of veratridine action on crustacean nerve by different divalent cations

The action of veratridine on the resting membrane potential of intact crayfish axons in the presence of different extracellular alkaline-earth cations has been studied using intracellular microelectrodes. At extracellular pH values of 7, 6 and 5, the apparent binding sequence was consistently Ca 2+ ⩾Sr 2+ >Mg 2+ ≈Ba 2+. This particular selectivity sequence and the insensitivity of binding order to changes in extracellular pH have been noted for other biological preparations and provide structural information about the veratridine - interaction sites to which the alkaline earths bind.

Electrophysiological and pharmacological properties of glial cells associated with the medial giant axon of the crayfish with implications four neuron-glial cell interactions.

Schwann-like glial cells surrounding the medial giant axon of the crayfish (Procambarus clarkii) were impaled with glass microelectrodes to study their responses to cholinomimetics, cholinergic receptor blockers and ouabain. Axon electrical properties were simultaneously monitored. Glial cells have a low membrane potential compared to the axon; -42 mV and -85 mV, respectively. Acetylcholine, carbachol and nicotine hyperpolarized the glial cells but did not affect the axon steady-state or active membrane potentials. The action of the cholinergics was completely blocked by d-tubocurarine and alpha-bungarotoxin. Ouabain hyperpolarized the glial cell but depolarized the axon. Tubocurarine blocked the ouabain hyperpolarization but not the delayed depolarization of the glia cell or the axon. It is concluded that ouabain causes the release of acetylcholine from the glial cell-axon preparation, inducing the glial hyperpolarization. Studies of the axon-glial cell interaction suggest that a function of the glial cell is to actively modulate the periaxonal potassium concentration on a signal from the axon. Periaxonal potassium can strongly affect axon membrane potential through electrogenic Na transport, modifying axon signalling properties.

Membrane potential of squid axons: effect of internal and external ion concentration.

Transmembrane resting potentials of squid axons were measured with variation of intracellular salt concentrations. It was found that the dependence of resting potential on the extracellular K-ion concentration was practically unaffected by the change of ionic strength in the intracellular phase, and the relationship of membrane potential and the logarithm of extracellular K-ion concentration was a straight line with a slope of about 50 mV/10-fold extracellular K-ion concentration. Also, the degree of change in resting potential due to the change of intracellular K-ion concentration was not affected appreciably by the extracellular solution with various K-ion concentrations, and the membrane potentials were insensitive over a wide range of intracellular K-ion concentrations. It was also indicated that the observed transmembrane potentials are not explained by a conventional membrane potential theory.

Action of extracellular pH on Na+ and K+ membrane currents in the giant axon of Loligo vulgaris.

Voltage-clamp currents and resting membrane potential of squid giant axons have been studied at extracellular pH varying between 4 and 10. The membrane currents, analyzed according to the Hodgkin-Huxley equations, showed that sodium permeability, PNa(E), and potassium conductance gK(E), curves were shifted toward positive voltages by different amounts and slightly depressed as the external pH was lowered. Under the same conditions, taum(E) and taun(E) were found to be enhanced and shifted to a larger extent in the same direction. The rate constants alpham and alphan were shifted substantially toward positive voltages, but betam and betan changed hardly at all. The shift of the alpham(E) curve was analyzed in terms of a fixed surface charge model; it indicates that unspecific negative groups with an approximate pKa of 4.5 are located in the vicinity of sodium active sites with an average charge separation of 8 A. A similar figure is obtained for the potassium system from the shift of the alphan(E) curve.

Sustained depolarizing potentials in reticulospinal axons during evoked seizure activity in lamprey spinal cord.

1. Intracellular recordings were made from lamprey reticulospinal axons (Müller axons) during seizures evoked by electrical stimulation of the isolated spinal cord in saline containing either 0 Cl or 1 mM picrotoxin. The seizures had tonic and clonic-phases similar to ictal seizures in mammalian brain. 2. During seizures Müller axons were depolarized by 10-15 mV. These seizure-depolarizations were not due to any direct effect of the evoking stimulus on the Müller axons themselves nor were they initiated by an accumulation or extracellular potassium. 3. A decrease in axonal input resistance occurred during a seizure-depolarization. Also, the amplitude of a seizure-depolarization was decreased by depolarizing the axon 5-15 mV with injected current. Further, hyperpolarizing the axon increased the amplitude of the seizure-depolarization, but the growth flattened out beyond 30-40 mV of hyperpolarization. The decrease in input resistance during the seizure-depolarization and the dependence of the response amplitude on axonal membrane potential suggested that the seizure-depolarization was an excitatory synaptic potential. However, the failure of the seizure-depolarization amplitude to continue to grow at membrane potentials greater than 30 mV negative to the resting potential was not consistent with this interpretation. 4. A synaptic conductance change as the cause of the seizure-depolarization was ruled out by setting the axonal membrane potential at different levels with injected current and monitoring the input resistance of the axon before and during seizure-depolarizations. It was found that no change in input resistance occurred during the seizure-depolarization when the axon was hyperpolarized more than approximately 30 mV, the same potential at which the growth in the response amplitude ceased. From analysis of these data and the passive current-voltage properties of Müller axons it is concluded that the seizure-depolarization is not a chemical synaptic potential, but rather the result of the passive injection of depolarizing current into the axons. 5. The source of the depolarizing current which flows into Müller axons during seizures is probably paroxysmal action-potential activity in spinal motoneurons and interneurons, many of which are electrically coupled to Müller axons.

Methylmercury: Effects on electrical properties of squid axon membranes

At low concentrations (25–100 μM) methylmercury chloride caused a steady increase in the threshold for excitation and on eventual block of action potentials without changing the resting membrane potential in squid giant axons. In the axons exposed to 25 μM methylmercury chloride, peak transient and steady-state conductances were decreased by 58.8 ± 5.1% and 35.9 ± 4.3% (mean ± SEM, 4 axons), respectively and leakage conductance increased to about five times of the control value. Higher concentrations of methylmercury chloride decreased the resting membrane potential. A concentration of 0.5 mM depolarizing the nerve membrane by 16 ± 2 mV (mean ± SEM, 3 axons) in 40 minutes. These changes in ionic conductances and membrane potential were irreversible on washing the axon with drug-free sea water.

Grayanotoxin, veratrine, and tetrodotoxin-sensitive sodium pathways in the Schwann cell membrane of squid nerve fibers.

The actions of grayanotoxin I, veratrine, and tetrodotoxin on the membrane potential of the Schwann cell were studied in the giant nerve fiber of the squid Sepioteuthis sepioidea. Schwann cells of intact nerve fibers and Schwann cells attached to axons cut lengthwise over several millimeters were utilized. The axon membrane potential in the intact nerve fibers was also monitored. The effects of grayanotoxin I and veratrine on the membrane potential of the Schwann cell were found to be similar to those they produce on the resting membrane potential of the giant axon. Thus, grayanotoxin I (1-30 muM) and veratrine (5-50 mug-jl-1), externally applied to the intact nerve fiber or to axon-free nerve fiber sheaths, produce a Schwann cell depolarization which can be reversed by decreasing the external sodium concentration or by external application of tetrodotoxin. The magnitude of these membrane potential changes is related to the concentrations of the drugs in the external medium. These results indicate the existence of sodium pathways in the electrically unexcitable Schwann cell membrane of S. sepioidea, which can be opened up by grayanotoxin I and veratrine, and afterwards are blocked by tetrodotoxin. The sodium pathways of the Schwann cell membrane appear to be different from those of the axolemma which show a voltage-dependent conductance.

The influence of cardioactive steroids, metabolic inhibitors, temperature and sodium on membrane conductance and potential of crayfish giant axons.

1. The resting membrane potential and the current-voltage relation were measured in crayfish giant axons before and after treatment with cardioactive steroids, metabolic inhibitors, extracellular sodium depletion and low temperature. 2. The membrane resistance of axons treated with cardioactive steroids, metabolic inhibitors, and low extracellular sodium was reduced by 30-53% depending on the treatment. Low temperature also caused a decrease in the membrane resistance of the axon but the decrease was limited to potentials around the resting membrane potential. The temperature response of sodium depleted or ouabain treated axons was an increase in resistance at all points along the current-voltage relation. 3. All inhibitors and low temperature caused a depolarization of the membrane potential. Ouabain and strophanthidin were the most effective, reducing the membrane potential by an average of 9.6 mV in 10-20 min. Low sodium did not cause a depolarization but consistently reduced the membrane resistance by an average of 30%. 4. The data suggest that there is an interaction between the activity of the ouabain-sensitive transport system and resting membrane resistance in the crayfish axon.

The influence of chloride on the ouabain-sensitive membrane potential and conductance of crayfish giant axons.

1. Resting potential and current-voltage relation were measured in crayfish giant axons bathed in chloride-free and sodium-free solutions with and without ouabain. 2. Chloride-free solution caused a transient depolarization but did not alter the steady-state membrane potential. Utilizing isethionate as an anion substitute, the membrane resistance increased 12.5%. 3. In the absence of extracellular chloride, ouabain (0.5-1 mM) depolarized the axon 6-7 mV. The shape of the current-voltage relation did not change but the curve was shifted along the current axis. 4. These results indicate that ouabain inhibits a steady-state hyperpolarizing electrogenic pump current of approximately 3 muA/cm2. 5. Extracellular sodium removal from axons equilibrated in chloride-free solutions transiently hyperpolarized the membrane 6-7 mV without a change in membrane resistance. The transient hyperpolarization was ouabain and temperature sensitive. The steady-state potential reached in sodium-free and chloride-free solution was not ouabain sensitive. Temperature sensitivity of the steady-state membrane potential was greatly reduced. 6. The transient hyperpolarization produced by extracellular sodium removal was metabolically driven and may present the expression of a sodium efflux transport current of 7.0-7.5 muA/cm2. 7. Using electrophysiologically measured parameters, sodium and potassium conductance, influx and efflux currents and the coupling ratio for sodium/potassium transport are calculated from a modification of the conductance equation. 8. The sodium/potassium transport coupling ratio for steady-state conditions was estimated at 5:3 (1.67:1).

Local Anesthetic Activity of 1-(Morpholino or Piperidino)-2-Propanol Derivatives

The local anesthetic activities of l-(morpholino or piperidino)-2-propanol derivatives were investigated. These activities were observed with nerve trunk anesthesia, rabbit's corneal anesthesia and intradermal injection anesthesia. Among these propanol derivatives, No. 24 and No. 25 which possess thioether-type sulfur were the most active and the effective potency was enhanced by increasing the dissociation constant (pKa) value in a series of the compounds. Effects on cat spinal reflex were also observed. These propanol derivatives revealed no selective depression of presynaptic depression or depression of the potential of monosynaptic reflex (MSR) and polysynaptic reflex (PSR). Procaine and Iidocaine had similar actions. Effects on axonal membrane potential and interaction of the calcium on these derivatives were also investigated. These derivatives decreased the action potential without altering the resting membrane potential and when antagonized with calcium, sciatic nerve action potential was decreased. Procaine and Iidocaine also showed the same results.

Effects of acetylcholine and carbamylcholine on the axon and Schwann cell electrical potentials in the squid nerve fibre.

1. The effect of acetylcholine and carbamylcholine on the axon and Schwann cell membrane potential have been studied in the giant nerve fibre of the squid. The addition of carbamylcholine (10(-6)M) to the external sea-water medium has no appreciable effects on the resting and action potentials of the axon. However, it induces a long-lasting hyperpolarization in the surrounding Schwann cells of the unstimulated intact or slit nerve fibres which is completely blocked by D-tubocurarine (10(-9)M). Eserine (10(-9)M) prolongs the Schwann cell hyperpolarizations induced by a 1 min exposure of the unstimulated nerve fibres to acetylcholine (10(-7)M).2. The addition of carbamylcholine (10(-6)M) to the external medium increases the relative permeability of the Schwann cell membrane to the potassium ion in slit nerve fibres. Yet, a hundredfold reduction in external sodium concentration has no appreciable effect on the hyperpolarization of the Schwann cells of the slit nerve fibre under similar conditions.3. Tetrodotoxin at a concentration of 5 x 10(-8)M has no appreciable effects on either the Schwann cell electrical potential or on the hyperpolarizing action of carbamylcholine on the Schwann cells of the unstimulated intact nerve fibres.4. These findings indicate the presence of acetylcholine receptors in the plasma membrane of the Schwann cell in these nerve fibres and give further support to the hypothesis on the role of the cholinergic system in the genesis of the long-lasting Schwann cell hyperpolarizations caused by the conduction of nerve impulse trains by the axon.

Low-impedance capillary electrode for wide-band recording of membrane potential in large axons.

An electrically floating platinized-platinum (Pt-Pt) wire within the length of a long (3-cm) electrolyte-filled glass capillary (100? diam), which is used to monitor the internal potential in large axons, eliminates cable effects and lowers the effective electrode impedance to < 10 k?. With this low-source-impedance capillary electrode; absolute transmembrane potentials can be recorded and a fast-responding (0.5-?s) voltage-clamp system is obtained.