Voltage-dependent Ionic Conductances Research Articles

The Effect of Variations in Ionic Conductance Values on the Dynamics of a Mathematical Model of Non-Spiking A-Type Horizontal Cells in the Rabbit Retina

A previous study proposed a mathematical model of A-type horizontal cells in the rabbit retina. This model, which was constructed based on the Hodgkin-Huxley model, was described by a system of nonlinear ordinary differential equations. The model contained five types of voltage-dependent ionic conductances: sodium, calcium, delayed rectifier potassium, transient outward potassium, and anomalous rectifier potassium conductances. The previous study indicated that when the delayed rectifier potassium conductance had a small value, depolarizing stimulation could change the dynamic state of the model from a hyperpolarized steady state to a depolarized steady state. However, how this change was affected by variations in the ionic conductance values was not clarified in detail in the previous study. To clarify this issue, in the present study, we performed numerical simulation analysis of the model and revealed the differences among the five types of ionic conductances.

Tonic Dopamine Induces Persistent Changes in the Transient Potassium Current through Translational Regulation

Neuromodulatory effects can vary with their mode of transmission. Phasic release produces local and transient increases in dopamine (DA) up to micromolar concentrations. Additionally, since DA is released from open synapses and reuptake mechanisms are not nearby, tonic nanomolar DA exists in the extracellular space. Do phasic and tonic transmissions similarly regulate voltage-dependent ionic conductances in a given neuron? It was previously shown that DA could immediately alter the transient potassium current (I(A)) of identified neurons in the stomatogastric ganglion of the spiny lobster Panulirus interruptus. Here we show that DA can also persistently alter I(A), and that the immediate and persistent effects of DA oppose one another. The lateral pyloric (LP) neuron exclusively expresses type 1 DA receptors (D1Rs). Micromolar DA produces immediate depolarizing shifts in the voltage dependence of LP I(A), whereas tonic nanomolar DA produces a persistent increase in LP I(A) maximal conductance (G(max)) through a translation-dependent mechanism involving target of rapamycin (TOR). The pyloric dilator (PD) neuron exclusively expresses D2Rs. Micromolar DA produces an immediate hyperpolarizing shift in PD I(A) voltage dependence of activation, whereas tonic DA persistently decreases PD I(A) G(max) through a translation-dependent mechanism not involving TOR. The persistent effects on I(A) G(max) do not depend on LP or PD activity. These data suggest a role for tonic modulators in the regulation of voltage-gated ion channel number; and furthermore, that dopaminergic systems may be organized to limit the amount of change they can impose on a circuit.

Bang-bang optimality of energy efficient spikes in single neuron models

Event Abstract Back to Event Bang-bang optimality of energy efficient spikes in single neuron models Biswa Sengupta1*, Martin Stemmler2, Jeremy Niven1, Andreas Herz2 and Simon Laughlin1 1 University of Cambridge, United Kingdom 2 LMU Munich , BCCN Munich, Germany About 50-80% of the total energy allocated to the mammalian brain is used for signaling, mainly to drive the $Na^+$/ $K^+$ pump (Attwell & Laughlin, 2001). Due to the substantial contribution of action potentials to the energy consumption, the biophysical properties generating an action potential can be matched to make them energy efficient (Alle et al., 2009). By combining different voltage-dependent ionic conductances with varying biophysical properties, different neurons express a myriad of action potential shapes characterized by varying heights and widths (Bean, 2007). How widespread are energy efficient action potentials and what are the major factors defining the energy efficiency of a single action potential ? The passive electric properties of a neuron, namely its capacitance and input resistance, set a lower bound for the cost of a given action potential. Inward ( $Na^+$) and outward ($K^+$) voltage-dependent currents act in a push-pull manner to change the voltage; these currents will invariably overlap, which can cause the energetic cost to rise to more than eleven-fold the baseline cost. We use seven single compartment models to assess the energy cost of vertebrate (cerebellar granule neurons, cortical interneurons, thalamo-cortical relay neurons and hippocampal interneurons) and invertebrate (squid axons, crab axons and bee kenyon cells) action potentials. We show that the energy consumption of a single action potential is directly dependent on the overlap between the $Na^+$ and $K^+$ currents that generate them. By optimizing the ionic conductance parameters that affect this overlap, we investigate how close each neuron type is to the minimal energy expenditure. Using sensitivity analysis and perturbation theory for periodic systems we show that an optimal model minimizes the overlap between $Na^+$ and $K^+$ currents. Just as in optimal control theory, the ionic conductance parameters allow for a bang-bang control of the underlying $Na^+$ and $K^+$ currents eliciting the action potential. Energy efficient action potentials are neuron-dependent but generally have narrower width, shorter height and exhibit an extremely rapid, explosive onset. Our modeling shows that in some neurons, such as the thalamo-cortical relay neurons, the hippocampal GABAergic interneurons or fast-spiking cortical interneurons, the properties of the currents that minimize the energy cost of the action potential are close to the experimentally measured currents, suggesting selection for reducing the energy costs. The experimentally observed properties of the squid giant axon model however, consumes substantial energy when compared to the optimal model that consumes far lower energy. We suggest that the deviation between the parameters generating the theoretical minimum cost of the action potential and those observed in neurons is dependent upon the function of the neuron. Conference: Computational and Systems Neuroscience 2010, Salt Lake City, UT, United States, 25 Feb - 2 Mar, 2010. Presentation Type: Poster Presentation Topic: Poster session II Citation: Sengupta B, Stemmler M, Niven J, Herz A and Laughlin S (2010). Bang-bang optimality of energy efficient spikes in single neuron models. Front. Neurosci. Conference Abstract: Computational and Systems Neuroscience 2010. doi: 10.3389/conf.fnins.2010.03.00085 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 20 Feb 2010; Published Online: 20 Feb 2010. * Correspondence: Biswa Sengupta, University of Cambridge, Paris, United Kingdom, bs393@cam.ac.uk Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Biswa Sengupta Martin Stemmler Jeremy Niven Andreas Herz Simon Laughlin Google Biswa Sengupta Martin Stemmler Jeremy Niven Andreas Herz Simon Laughlin Google Scholar Biswa Sengupta Martin Stemmler Jeremy Niven Andreas Herz Simon Laughlin PubMed Biswa Sengupta Martin Stemmler Jeremy Niven Andreas Herz Simon Laughlin Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Local Generation of Theta-Frequency EEG Activity in the Parasubiculum

The parasubiculum is a major component of the hippocampal formation that receives inputs from the CA1 region, anterior thalamus, and medial septum and that projects primarily to layer II of the entorhinal cortex. Hippocampal theta-frequency (4-12 Hz) electroencephalographic (EEG) activity has been correlated with sensorimotor integration, spatial navigation, and memory functions. The present study was aimed at determining if theta is also generated locally within the parasubiculum versus volume conducted from adjacent structures. In urethan-anesthetized rats, the phase-reversal of theta activity between superficial and deep layers of the parasubiculum was demonstrated using differential recordings from movable bipolar electrodes that eliminate the influence of volume-conducted activity. Parasubicular theta was abolished by atropine, and was in phase with theta in stratum radiatum/lacunosum-moleculare of the CA1 region. Whole cell current-clamp recordings in brain slices were then used to determine if parasubicular theta may be generated in part by membrane potential oscillations in layer II neurons. Membrane potential oscillations occurred in most layer II neurons, including four putative interneurons, when cells were held at near-threshold voltages using current injection. The frequency of oscillations increased from 3.2 to 6.1 Hz when bath temperature was raised from 22 to 32 degrees C, and oscillations persisted in the presence of blockers of fast ionotropic glutamatergic and GABAergic synaptic transmission. Oscillations are therefore likely generated by intrinsic, voltage-dependent ionic conductances. These results indicate that theta field activity is generated locally within the parasubiculum and that intrinsic membrane potential oscillations, synchronized by local inhibitory inputs, may contribute to the generation of this activity.

Voltage-dependent ionic conductances in the human malignant astrocytoma cell line U87-MG.

Although the human malignant astrocytoma cell line U87-MG has been used in numerous studies, few findings are available on the properties of its membrane ion conductances. Characterization of the ion channels expressed in these cells will make it possible to study membrane ion conductance changes when a receptor is activated by its ligand. This will help to elucidate the functional properties of these receptors and their signal-transduction pathways in pathophysiological events. This work studied the voltage-dependent ionic conductances of U87-MG cells using the Whole-Cell Recording patch-clamp technique. Six types of voltage-dependent ionic currents were identified: (i) a TEA-, 4-AP- and CTX-sensitive Ca2+-dependent K+ current, (ii) a transient K+ current inhibited by 4-AP, (iii) an inwardly rectifying K+ current blocked by Ba2+ and 4-AP, (iv) a DIDS- and SITS-sensitive Cl- current, (v) a TTX-sensitive Na+ conductance and (vi) a L-type Ca2+ conductance activated by BayK-8644 and inhibited by Ni and the L-type Ca2+ channel inhibitor, nifedipine. In addition, electrical depolarizations elicited inward currents due to voltage-independent, Ca2+-dependent K+ influx against the electrochemical gradient, probably via an ouabain-sensitive Na+-K+ pump.

A Compartmental Model of an External Urethral Sphincter Motoneuron of Onuf’s Nucleus

This article discusses a model of the electrical behavior of an external urethral sphincter motoneuron, based on morphological parameters like soma size, dendritic diameters and spatial dendritic configuration, and several electrical parameters. Because experimental data about the exact ion conductance mix of external urethral sphincter neurons is scarce, the gaps in knowledge about external urethral sphincter motoneurons were filled in with known data of a-motoneurons. The constructed compartmental model of motoneurons of Onuf’s nucleus contains six voltage-dependent ionic conductances: a fast sodium and potassium conductance and an anomalous rectifier in the soma; a fast delayed rectifier type potassium conductance and a fast sodium conductance in the initial axon segment; an L-type calcium channel in the dendritic compartments. This paper considers the simulation of external urethral sphincter motoneuron responses to current injections that evoke bistable behavior. Simulations show self-sustained discharge following a depolarizing pulse through the microelectrode; the firing was subsequently terminated by a short hyperpolarizing pulse. This behavior is highly functional for neurons that have to exhibit prolonged activation during sphincter closure. In addition to these ’on’ and ’off ’ responses, we also observed a particular firing behavior in response to long-lasting triangular current pulses. When the depolarizing current was slowly increased and then decreased (triangular pulse) the firing frequency was higher during the descending phase than during the initial ascending phase.

Tricyclic anticonvulsants: mechanisms of action

Two aspects of neuronal function are particularly relevant to the known actions of antiepileptic drugs (AEDs): voltage-dependent ionic membrane conductances (especially sodium, potassium, and calcium); and synaptic actions of neurotransmitters, particularly c-aminobutyric acid (GABA), glutamate, and acetylcholine. Inhibitory actions in the central nervous system (CNS) are mediated principally by GABA. Other neurotransmitters, including norepinephrine, dopamine, and serotonin, also have inhibitory properties [1]. GABA receptors, which have been cloned, are composed of a GABAA receptor, a barbiturate receptor, and a benzodiazepine receptor, which all surround the chloride channel [2]. Neurons that secrete the excitatory amino acid L-glutamate are ubiquitous throughout the CNS. Cholinergic neurons extend from the septum and basal forebrain to the cortex, including the hippocampus. Recent evidence suggests a role for acetylcholine in epileptogenesis; e.g., excised temporal lobectomy tissues show increased concentrations of acetylcholinesterase compared with what is expected [1].

Energy efficient modulation of dendritic processing functions

The voltage dependent ionic conductances and the passive properties of the neural membrane determine how external inputs are processed by the dendritic tree, and define the computational characteristics of neurons. However, what controls these characteristics and how they are implemented at the single neuron level, in such a way that an external input results in the coding of the appropriate output, is essentially unknown. We show here that a slow inactivation of the Na + channel, involved in the attenuation and/or failure of APs in the dendrites, acts as an active and energy efficient filter of synaptic input, and results in an activity-dependent control of the properties of individual neurons. Thus, the activation or expression of this mechanisms could be an efficient way to selectively modulate the input/output processing properties of dendrites, and could be needed to limit or suppress the onset of a number of pathological brain disorders.

S-8-1 - State-dependent regulation of neuronal excitability by dopamine

Since the discovery that the loss of the dopaminergic innervation of the striatum resulted in Parkinson's disease, physiologists have attempted to understand the role of dopamine on striatal activity. Hypotheses relying upon concepts derived from studies of fast synaptic transmission have consistently failed to explain the actions of dopamine or other receptors coupled to G-proteins which modulate the properties of voltage-dependent ionic conductances responsible for synaptic integration and spike activity. Recently, patch clamp studies have revealed that in medium spiny striatal neurons dopamine D1-class receptors modulate voltage-dependent Na+, K+ and Ca2+ channels. From a consideration of the biophysical properties of these channels and the state transitions that medium spiny neurons undergo while responding to cortical input, a novel picture of dopamine's actions is beginning to emerge. Our results and those of others suggest that D2-class receptors serve to make the transition to the depolarized 'upstate' from the hyperpolarized 'downstate' more probable in response to cortical input. But, once the transition has occurred, the alteration in excitability should be short-lived unless the neuron has recently been active. This state-dependent modulation provides a mechanism by which dopamine could shape global striatal activity governing the execution of motor behaviors.

Pharmacological induction of rhythmical activity and plateau action potentials in unmyelinated axons

The physiological function of the axon is to conduct short all-or-none action potentials from their site of initiation (usually the cell body) to the synapse. To ensure this function, both passive and active biophysical properties of the axons are tuned very precisely, especially the voltage-dependent ionic conductances to sodium and potassium. Under normal conditions, axons are not spontaneously active. Minor modifications of their ionic micro-environment or slight changes in the membrane properties are however sufficient to induce rhythmical activity and modify the time course of the action potentials. These modifications can be induced by a variety of pharmacological agents. Some typical examples taken from original studies on invertebrate preparations are illustrated. The experiments were carried out on two axonal preparations: the giant axon of the squid Loligo forbesi and the giant axon of the cockroach Periplaneta americana. The axons were ‘space-clamped’ and studied under both current-clamp and voltage-clamp conditions. Voltage-clamp experiments were used to dissect out the mechanisms underlying repetitive activity and to extract the relevant parameters. These parameters were then used to rebuild the observed effects using an extended version of the Hodgkin and Huxley (1952, J Physiol (Lond) 117, 500–544) formulation. One easy way to get repetitive firing in both preparations is to reduce potassium conductance. The effect of 4-aminopyridine on squid axon is illustrated here. The experimental results, including the occurrence of bursts of activity, can be described by adding a time- and voltage-dependent block of the potassium channels to the original Hodgkin and Huxley (1952, J Physiol (Lond) 117, 500–544) model. Repetitive spike activity and plateau action potentials are also produced when the depolarising effect of the voltage-dependent potassium current is counterbalanced by a maintained inward sodium current. This maintained sodium current can be due to several different mechanisms. This will be illustrated by five structurally unrelated molecules: two scorpion toxins, two insecticide molecules and one sea anemone toxin. One toxin purified from the venom of the scorpion Buthotus judaïcus (insect toxin 1) exerts its effects by shifting the sodium activation curve towards more hyperpolarized potentials. Another toxin purified from the venom of another scorption Androctonus australis (mammal toxin 1) modifies a significant proportion of normal (fast) sodium channels into slowly activating and inactivating sodium channels. The main effect of the insecticide DDT is to maintain sodium channels in the ‘open’ configuration. Another insecticide molecule known to induce repetitive activity, S-bioallethrin, activates voltage-dependent sodium channels with slow activation and inactivation kinetics. The sea anemone toxin anthopleurin A, purified from the venom of Anthopleura xanthogrammica, delays inactivation of the sodium current without changing its activation kinetics. These examples show that minor modifications of the properties of the nerve membrane are sufficient to alter nerve function. These deleterious effects will be amplified at the synapse through dramatic changes in transmitter release and will lead eventually to disastrous alterations of brain function.

Voltage-dependent ionic conductances in Chinese hamster ovary cells.

Chinese hamster ovary (CHO) cells are becoming a widely used biological material. A number of studies report membrane ion conductance changes after transfection of channels and receptors, but there are few data available on the properties of membrane ion conductances of CHO cells before transfection. In this work we studied voltage-dependent ionic conductances in cultures of CHO native (CHO-K1) cells. Three types of voltage-dependent ionic conductances were identified: 1) a K+ conductance showing sensitivity to Ca2+ and a unit conductance of approximately 210 pS in symmetrical 150 mM K+ outside-out patches (this conductance, which did not inactivate during a 160-ms pulse, was inhibited by 30 nM charybdotoxin but not by 30 mM extracellular tetraethylammonium); 2) a rapidly activating and inactivating tetrodotoxin (TTX)-sensitive inward current, peaking at about -10 to 0 mV (this current showed characteristics similar in many respects to Na+ current recorded in neurons); and 3) another voltage-dependent inward current, which had slow inactivation, was TTX insensitive but was blocked by Co2+ (current was also carried by Ba2+, peaked at approximately 0 to +10 mV, was identified as a Ca2+ conductance, and was inhibited by dihydropyridines but not by 10 microM omega-conotoxin). Cell-attached patch recordings of single Ca2+ channel currents demonstrated a unitary conductance of approximately 20 pS.

Voltage-dependent ionic conductances of type I spiral ganglion cells from the guinea pig inner ear

Type I spiral ganglion cells provide the afferent innervation to the inner hair cells of the mammalian organ of Corti and project centrally to the cochlear nucleus. While single-unit studies conducted over the past several decades have provided a wealth of information concerning the response characteristics of these neurons and, to some extent, their receptor targets, little is known about the neuron's intrinsic electrical properties. These properties undeniably will contribute to the firing patterns induced by acoustic stimuli. Type I spiral ganglion cell somata from the guinea pig inner ear were acutely isolated and the voltage-dependent conductances were analyzed with the whole-cell voltage clamp. Under conditions that mimic the normal intra- and extracellular ionic environments, type I spiral ganglion cells demonstrate fast inward TTX-sensitive Na currents (whose current density varied markedly among cells) and somewhat more slowly developing outward K currents. Resting potentials averaged -67.3 mV. Under current clamp, no spontaneous spike activity was noted, but short current injections produced graded action potentials with after hyperpolarizations lasting several milliseconds. The nondecaying outward K current activated at potentials near rest and was characterized by a pronounced rectification. The kinetics of the Na and K currents were rapid. Maximum peak inward Na currents occurred within 400 microseconds, between a voltage range of -10 and 0 mV, and inactivated within 4 msec. Recovery from inactivation was also rapid. At a holding potential of -80 mV, the time constant for recovery from an inactivating voltage step to -10 mV was 2.16 msec. Above -50 mV outward K currents reach half-maximal amplitude within 1.5 msec. In addition to these currents, a slow noninactivating TTX-sensitive inward current was observed that was blockable with Cd2+ or Gd3+. Problems encountered with blocking the tremendous outward K current hampered the characterization of this inward current. Similarities between the kinetics of ganglion cell currents and some of the rapid temporal characteristics of eighth nerve single-unit activity confirm the notion that intrinsic membrane properties help shape auditory neuron responses to sound.

Synaptic and intrinsic conductances shape picrotoxin-induced synchronized after-discharges in the guinea-pig hippocampal slice.

1. A computer model was constructed of the guinea-pig hippocampal region in vitro, containing 100 pyramidal neurones. This approach has contributed to the understanding of brief (usually less than 100 ms) epileptic events known as 'interictal spikes'. The present study addresses the cellular mechanisms of more prolonged epileptic events, lasting 200 ms and more, that may represent short-duration seizures. Each neurone was simulated with a nineteen-compartment model using six voltage-dependent ionic conductances. The neurones were randomly interconnected with excitatory synapses, each synapse exerting a fast voltage-independent alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) component and a slower voltage-dependent N-methyl-D-aspartate (NMDA) component. Each neurone received input from twenty other neurones. 2. This model was able to generate, in response to synaptic noise or to stimulation of one neurone, a series of synchronized population bursts, the initial (primary) burst being longer than later (secondary) bursts, terminating in a prolonged after-hyperpolarization. The simulated after-discharge potentials resemble those recorded experimentally from pyramidal neurones during perfusion of the hippocampal slice with media containing picrotoxin, a blocker of synaptic inhibition mediated by GABAA receptors. 3. Simulated after-discharges agree with the following experiments: over a certain range of total NMDA conductance, blockade of AMPA receptors will prevent the occurrence of synchronized firing, whereas, blockade of NMDA receptors will, in contrast, abolish the secondary bursts, leaving a shortened and somewhat smaller primary burst. Dendritic potential oscillations occur in phase with somatic oscillations. When interneurones (some generating GABAA-mediated IPSPs, others generating GABAB IPSPS) are included in the model, the occurrence of synchronized events was suppressed, the most significant suppressant effect coming from GABAA IPSPS. 4. The model predicts that: a dendritic calcium spike occurs during each secondary burst; AMPA receptors serve to maintain the synchrony of secondary bursts, as well as to initiate the primary burst; and that with sufficient total NMDA conductance, synchronized firing can occur even with AMPA receptors blocked. 5. The model suggests, in addition, that the duration of the initial burst is determined in part by the experimentally observed delay between Ca2+ entry and peaking of the after-hyperpolarization (AHP) conductance, and hence reflects properties of the individual pyramidal neurones. Specifically, a pattern of a long initial burst followed by brief secondary bursts is elicited in single-cell simulations by injection of a steady depolarizing current into the apical dendrite. The same pattern is produced when the single-cell model includes only calcium and calcium-dependent conductances.(ABSTRACT TRUNCATED AT 400 WORDS)

Voltage-Clamp Frequency Domain Analysis of Nmda-Activated Neurons

1. Voltage and current-clamp steps were added to a sum of sine waves to measure the tetrodotoxin-insensitive membrane properties of neurons in the intact lamprey spinal cord. A systems analysis in the frequency domain was carried out on two types of cells that have very different morphologies in order to investigate the structural dependence of their electrophysiological properties. The method explicitly takes into account the geometrical shapes of (i) nearly spherical dorsal cells with one or two processes and (ii) motoneurons and interneurons that have branched dendritic structures. Impedance functions were analysed to obtain the cable properties of these in situ neurons. These measurements show that branched neurons are not isopotential and, therefore, a conventional voltage-clamp analysis is not valid. 2. The electrophysiological data from branched neurons were curve-fitted with a lumped soma-equivalent cylinder model consisting of eight equal compartments coupled to an isopotential cell body to obtain membrane parameters for both passive and active properties. The analysis provides a quantitative description of both the passive electrical properties imposed by the geometrical structure of neurons and the voltage-dependent ionic conductances determined by ion channel kinetics. The model fitting of dorsal cells was dominated by a one-compartment resistance and capacitance in parallel (RC) corresponding to the spherical, non-branched shape of these cells. Branched neurons required a model that contained both an RC compartment and a cable that reflected the structure of the cells. At rest, the electrotonic length of the cable was about two. Uniformly distributed voltage-dependent ionic conductance sites were adequate to describe the data at different membrane potentials. 3. The frequency domain admittance method in conjunction with a step voltage clamp was used to control and measure the oscillatory behavior induced by N-methyl-D-aspartate (NMDA) on lamprey spinal cord neurons. Voltage-clamp currents and impedance functions were measured at different membrane potentials. The impedance functions had a voltage-dependent resonance and phase shift characteristic of a negative conductance. These measurements provide a quantitative analysis of the conductances induced by NMDA in central neurons of the lamprey spinal cord and directly establish the basis of the non-linear oscillatory behavior previously observed in the presence of NMDA. NMDA was shown specifically to activate a negative and a positive conductance, both of which were markedly affected by the membrane potential. It is shown that the net current in the presence of NMDA must be considered as the algebraic sum of currents in opposite directions.(ABSTRACT TRUNCATED AT 400 WORDS)

Voltage-gated currents of putative GABAergic amacrine cells in primary cultures and in retinal slice preparations

To analyze the voltage-dependent ionic conductances of putative GABAergic amacrine cells developing in vitro, whole cell patch clamp recordings were carried out on identified neurons in monolayer cultures from embryonic chick retinae. These recordings were directly compared with those performed on amacrine cells in chick retinal slice preparations. Current responses to depolarizing voltage steps observed in cultured neurons could be separated into at least four different components. A small tetrodotoxin-sensitive sodium inward current was observed in approximately 50% of the cells. The considerably larger outward pottassium current consisted of a transient 4-aminopyridine-sensitive component and a sustained component. The latter was reduced in the presence of both tetraethylammonium chloride and Co 2+ and thus was probably composed of two conductances. In addition, a Ca 2+-carried inward current of small amplitude could be identified. Voltage-sensitive currents measured in amacrine cells of retinal slices were very similar. Again, only about half of the cells exhibited sodium currents. Potassium currents contained the above components, but their contributions to the whole cell current seemed to be different. Together with previous findings these results suggest that immature retinal neurons in dissociated cultures undergo a differentiation process similar to that occuring in vivo.

Ionic currents of the lateral pyloric neuron of the stomatogastric ganglion of the crab

1. The lateral pyloric (LP) neuron is an important component of the network that generates the pyloric rhythm of the stomatogastric ganglion (STG) and is a direct target of many modulatory inputs to the STG. Our aim in this and the subsequent two papers is to describe the conductances present in this cell and to understand the role these conductances play in shaping the activity of the neuron. 2. LP neurons were studied in two-electrode voltage clamp (TEVC) in a saline solution containing tetrodotoxin (TTX) and picrotoxin (PTX) to isolate them pharmacologically from presynaptic inputs. 3. We identified six voltage-dependent ionic conductances. These include three outward currents that resemble a delayed rectifier current, a Ca(2+)-activated K+ current and an A-current similar to those seen in many other preparations. LP neurons show three inward currents, a fast TTX-sensitive current, a hyperpolarization-activated inward current, and a Ca2+ current.

Action of heptaminol hydrochloride on contractile properties in frog isolated twitch muscle fibre

1. Heptaminol stopped or delayed the progressive decline in tension which characterizes the phenomenon of fatigue in frog isolated twitch muscle fibre. 2. Heptaminol had no action on the sodium, potassium and calcium voltage-dependent ionic conductances. 3. The hypothesis of an action via an internal alkalinization was tested by comparison with the action of NH4Cl. Both substances increased the tension. 4. The action of heptaminol was suppressed in sodium-free (TRIS) solution or in the presence of amiloride while the action of NH4Cl was always observed. 5. These results could be explained by a stimulation of the Na/H antiport by heptaminol.

Neuron simulations with SABER

Computational models can provide critical tests of hypotheses of neuronal function. These models are essential for dealing with the complications of time- and voltage-dependent (active) ionic conductances. Commercial circuit analysis programs have been useful tools for this work. We report our experience modelling biophysically realistic membrane properties with SABER (Analogy, Inc.), a new general purpose simulator. SABER allows construction of models with arbitrary membrane properties. This is a major advantage over similar programs (e.g. SPICE), which are limited to a predefined library of electronic components. The empirically determined equations that describe rate constants, ionic conductances, currents, and concentration shifts can be translated directly into model elements (‘templates’) written in C-like code. We describe the development of SABER models that simulate a synapse and an action potential.

Studies of solitary semicircular canal hair cells in the adult pigeon. II. Voltage-dependent ionic conductances

1. The ionic conductances present in putative type II hair cells enzymatically dissociated from the anterior, posterior, and lateral semicircular canal cristae of the white king pigeon (Columba livia) vestibule were studied under whole cell voltage clamp. 2. Two classes of voltage-dependent potassium conductances were distinguishable on the basis of the time course of activation and inactivation and pharmacologic sensitivity. The rapid potassium conductance, IA, as inhibited by 6 mM 4-aminopyridine (4-AP), whereas the slow potassium conductance, IK, was inhibited by 50 mM tetraethylammonium (TEA). These conductances were not affected by extracellular calcium removal. IA was quite similar to the rapidly-inactivating A-current of molluscan soma, whereas IK was more like the delayed rectifier of molluscan soma. 3. The steady-state inactivation of IA occurred over a potential range from -100 to -40 mV. The threshold for activation of IA occurred between -60 and -50 mV. The slope conductance of the I-V curve over a range of -50 to -20 mV was 13.7 nS when the conditioning pulse was -100 mV, and we estimate it to be approximately 1-2 nS from the resting membrane potential of -56 mV. 4. The steady-state inactivation of IK was approximately 60% at -40 mV and was completely removed at -80 mV. The threshold for activation of IK was between -50 and -40 mV. The slope conductance of the I-V curve over a range of -50 to -20 mV was 10.5 nS when the conditioning pulse was -80 mV, and we estimate it to be approximately 6-7 nS from the resting potential of -56 mV. 5. At -56 mV (the average resting membrane potential of putative type II semicircular canal hair cells), approximately 10-14% of IA channels and approximately 57-70% of IK channels were not inactivated: thus IA and IK can contribute to the outward current during small depolarizations from rest. 6. A small calcium-dependent outward current, IK(Ca), could be elicited during step depolarizations from a holding potential of -40 mV. This calcium-dependent current was active over the range of -20 to +40 mV. 7. Inward currents could not be detected when the cells were exposed to normal physiological solutions. However, when the outward currents were blocked with internal cesium and the external solution contained 20 mM barium, sustained inward currents with rapid activation kinetics could be detected. The threshold for activation of the inward current occurred at -40 mV, and the I-V relationship peaked at -10 mV.(ABSTRACT TRUNCATED AT 400 WORDS)

Voltage clamping with single microelectrodes: Comparison of the discontinuous mode and continuous mode using the Axoclamp 2A amplifier

The voltage clamp technique is a powerful method for studying the physiology of excitable membrane. This technique has made possible the determination of ionic responses generated by activation of either receptor-mediated or voltage-dependent processes. The development of the whole-cell, 'tight-seal' voltage clamp method has allowed the analysis and examination of membrane physiology at the single cell level. The method allows the characterization of voltage-dependent ionic conductances both at the macroscopic (whole-cell) and at the microscopic (unitary conductance or single channel) level in cells less than 10 micron in diameter, a feat difficult to achieve with 'conventional' fine-tipped micropipettes. In this paper, several methologies used for culturing neuronal and non-neuronal cells in the laboratory are described. A comparison between the two modes of voltage clamp using blunt-tipped 'patch'-microelectrodes, the switching (discontinuous) and the non-switching (continuous) modes, of the Axoclamp-2A amplifier is made. Some results on membrane currents obtained from neuronal and non-neuronal cells using the single electrode whole-cell 'tight-seal' voltage clamp is illustrated. The possible existence of two inactivating K+ currents, one dependent on Ca++ the other is not, is discussed.