Intrinsic Electrophysiological Properties Research Articles

Adenosine promotes burst activity in guinea-pig geniculocortical neurones through two different ionic mechanisms.

1. The mechanisms of action of adenosine were examined in relay neurones of the dorsal lateral geniculate nucleus (LGND) using in vitro intracellular recording techniques in guinea-pig thalamic slices. 2. Adenosine hyperpolarized LGND relay neurones due to an increase in membrane potassium conductance. The K+ currents generated by near maximal stimulation of adenosine and GABAB receptors were non-additive. 3. Blockage of membrane K+ conductances by barium unmasked a second response to adenosine; an outward shift of the current versus voltage relationship negative to -65 mV associated with an increase in membrane input resistance. The beta-adrenoceptor agonist isoprenaline elicited an inward current in the same voltage range, which was inhibited and replaced by an outward current during activation of adenosine receptors. The effects of adenosine were due to a decrease in amplitude and rate of rise of the hyperpolarization-activated cation current, Ih. Maximal reduction by 66% of Ih amplitude occurred near the range of half-activation. 4. Both responses to adenosine were mimicked by the selective A1 receptor agonists N6-cyclopentyladenosine or N6-cyclohexyladenosine, and reversibly blocked by the selective A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). 5. The decrease in Ih by adenosine may be mediated by an inhibition of adenylyl cyclase activity and hence a decrease in the intracellular level of cyclic AMP, since local application of the adenylyl cyclase inhibitor 2',3'-dideoxyadenosine imitated the decrease in Ih. Local application of the adenylyl cyclase stimulant forskolin or 8-bromo-cyclic AMP resulted in an enhancement in Ih, and forskolin inhibited the action on Ih evoked by N6-cyclopentyladenosine. 6. The adenosine-induced effects interacted with the intrinsic electrophysiological properties of LGND neurones in that (i) the hyperpolarization due to an increase in K+ conductance inhibited single spike firing and promoted calcium-mediated burst discharges, and (ii) the decrease in Ih inhibited the dampening effect on Ca(2+)-mediated rebound activity of beta-adrenergic receptor stimulation. 7. It is suggested that during increased levels of extracellular adenosine the response of LGND relay neurones to activating brainstem influences will be depressed, and a pattern of Ca(2+)-mediated burst firing will be favoured.

Serotonin hyperpolarizes cholinergic low-threshold burst neurons in the rat laterodorsal tegmental nucleus in vitro.

Serotonergic suppression of cholinergic neuronal activity implicated in the regulation of rapid eye movement sleep and its associated phenomenon, pontogeniculooccipital waves, has long been postulated, but no direct proof has been available. In this study, intracellular and whole-cell patch-clamp recording techniques were combined with enzyme histochemistry to examine the intrinsic electrophysiological properties and response to serotonin (5-HT) of identified cholinergic rat laterodorsal tegmental nucleus neurons in vitro. Sixty-five percent of the recorded neurons demonstrated a prominent low-threshold burst, and of these, 83% were cholinergic. In current-clamp recordings 64% of the bursting cholinergic neurons tested responded to the application of 5-HT with a membrane hyperpolarization and decrease in input resistance. This effect was mimicked by application of the selective 5-HT type 1 receptor agonist carboxamidotryptamine maleate. Whole-cell patch-clamp recordings revealed that the hyperpolarizing response was mediated by an inwardly rectifying K+ current. Application of 5-HT decreased excitability and markedly modulated the discharge pattern of cholinergic bursting neurons: during a 5-HT-induced hyperpolarization these neurons exhibited no rebound burst after hyperpolarizing current input and a burst in response to depolarizing current input. In the absence of 5-HT, the relatively depolarized cholinergic bursting neurons responded to an identical hyperpolarizing current input with a burst and did not produce a burst after depolarizing current input. These data provide a cellular and molecular basis for the hypothesis that 5-HT modulates rapid eye movement sleep phenomenology by altering the firing pattern of bursting cholinergic neurons.

Synaptic transmission between individual pyramidal neurons of the rat visual cortex in vitro

Synaptic transmission between pairs of neurons in layer 2/3 of in vitro slices from the rat visual cortex was studied by dual intracellular recording. The intrinsic electrophysiological properties of these neurons suggested that they were pyramidal cells. More than 1/3 of the total number of synaptically connected neurons were stained by intracellular iontophoresis of biocytin, and all had pyramidal morphology. Postsynaptic potentials (PSPs) were evoked by single action potentials (elicited by current injection) in presynaptic cells. Measurements of PSP latency, amplitude, and shape were made on spike-triggered averages. Forty-eight synaptic connections were found out of a possible total of 549, equivalent to a probability of about 0.09. For these 48 connections, the distance between the 2 impalement sites ranged from 50 to 340 microns. All PSPs were depolarizing at rest (-74 +/- 5 mV, mean +/- SD), and all 14 of the PSPs that were also recorded with the postsynaptic cell depolarized to around action potential threshold (about -55 mV) remained depolarizing. PSPs had short latencies (1.2 +/- 0.6 msec, mean +/- SD), suggesting that they were mediated by monosynaptic pathways. Peak amplitudes of the averaged PSPs varied widely (range, 0.05-2.08 mV), but the majority were less than 0.5 mV. PSPs decayed exponentially with time constants that were correlated with, but slightly longer than, the membrane time constants of the postsynaptic cells measured using injected current pulses. Four cell pairs were connected reciprocally, and 6 examples of convergent input were found in which a single cell was postsynaptic to more than 1 presynaptic cell. Trial-to-trial fluctuations in PSP amplitude were analyzed for 16 synaptic connections. The amplitude of the PSP evoked by a presynaptic action potential fluctuated more than could be accounted for by the background noise.

The intrinsic electrophysiological properties of mammalian neurons: insights into central nervous system function.

This article reviews the electroresponsive properties of single neurons in the mammalian central nervous system (CNS). In some of these cells the ionic conductances responsible for their excitability also endow them with autorhythmic electrical oscillatory properties. Chemical or electrical synaptic contacts between these neurons often result in network oscillations. In such networks, autorhythmic neurons may act as true oscillators (as pacemakers) or as resonators (responding preferentially to certain firing frequencies). Oscillations and resonance in the CNS are proposed to have diverse functional roles, such as (i) determining global functional states (for example, sleep-wakefulness or attention), (ii) timing in motor coordination, and (iii) specifying connectivity during development. Also, oscillation, especially in the thalamo-cortical circuits, may be related to certain neurological and psychiatric disorders. This review proposes that the autorhythmic electrical properties of central neurons and their connectivity form the basis for an intrinsic functional coordinate system that provides internal context to sensory input.

The electroencephalographic pattern during electroconvulsive therapy: V. Observations on the origins of phase III delta energy and the mechanism of action of ECT.

The generation of the spike-wave activity of Phase III of ECT seizures is attributed to the recurrence of synchronized, prolonged periods of intense inhibitory current flow (hyperpolarization), and associated rebound spike bursts, produced by the inhibitory circuit relationships and intrinsic electrophysiological properties of thalamic neurons. An anatomical and neurophysiological model of the development of generalized, synchronous 3-Hz spike-wave seizure activity is proposed which outlines the origin, maintenance, slowing, and termination of this fundamental seizure rhythm. Phase III inhibitory current flow (delta energy) and/or spike bursts may bring about therapeutic benefit by initiating a chain of agonist-independent and agonist-dependent events which results in long-term augmentation of serotonergic and noradrenergic neurotransmission and diminution of cholinergic neurotransmission in the forebrain. A specific anatomical and functional model of the mechanism of action of ECT is proposed, in which: (1) adrenergic and cholinergic pathways in the forebrain are assumed to be massively stimulated during ECT seizures, whereas serotonergic pathways are assumed to be inhibited during these seizures; (2) the beneficial effects of ECT are considered to be more dependent upon ECT-induced changes in 5-HT neurotransmission than upon alteration of noradrenergic function; (3) these beneficial effects involve up-regulation of 5-HT2 and down-regulation of M1- and M2-muscarinic receptor densities by both agonist-independent and agonist-dependent mechanisms, coupled with functional augmentation of noradrenergic neurotransmission; and (4) these effects may be brought about by Phase III inhibitory current flow- and/or spike burst-induced alteration of the function of second-messenger generator systems.

Low-threshold calcium spikes in hypothalamic neurons recorded near the paraventricular nucleus in vitro

From guinea-pig hypothalamic slices, intracellular studies demonstrate the existence of neurons responding to depolarizing current pulses by bursts of fast spikes riding on slow depolarizing potentials, when activated at the resting potential or from hyperpolarized levels (44 cells). Slow depolarizing potentials have a mean amplitude of 17.6 mV and a mean duration of 65.2 msec. They are also produced at the termination of hyperpolarizing current pulses. The ionic basis for these slow potentials have been investigated. Fast spikes constituting the burst discharge are blocked by TTX but the slow component is unaffected, being blocked by Co ++ and enhanced by Ba ++. Taken together, these results show that the slow depolarizing potentials are generated by a low-threshold Ca ++ conductance which is de-inactivated by membrane hyperpolarization. When the neurons are spontaneously active, they exhibit bursts arising from slow depolarizing potentials reminiscent of those evoked by direct stimulation. They also show longer episodes of repetitive firing. Twelve neurons were intracellularly stained and were found in the periphery of the paraventricular nucleus (PVN), in close proximity to the groups of neurophysin-positive neuroendocrine neurons present in the lateral part of this nucleus. Injected neurons have the morphology of reticular cells, judging by their few multipolar, rectilinear and sparsely branched dendrites. Some of their processes are directed towards PVN. Because of their intrinsic electrophysiological properties and their possible relationships with PVN, the population of cells described in the present study may play a role in functions relating to the PVN.

Feasibility of electrical recordings from unconnected vertebrate CNS neurons cultured in a three-dimensional extracellular matrix

Single, i.e. unconnected spinal or cerebral neurons from chick embryos rapidly regenerate morphologically identifiable axons and dendrites when cultured in a three-dimensional (3-D) extracellular matrix (ECM) consisting of a hydrated native collagen lattice. We now show that it is possible to study the intrinsic electrophysiological properties of such neurons as early as 2 days in culture. Cells were plated at 10 5 cells/dish using Medium 199 containing 10% fetal calf serum but no other supplements or antibiotics-antimycotics. A patch-clamp/whole-cell voltage clamp system was used to record (1) single-channel currents from cell-attached patches, (2) transmembrane potentials during the injection of rectangular currents, and (3) whole-cell currents during voltage clamp. After 2–4 days in culture, isolated cerebral and spinal neurons exhibited single-channel currents. Within 7 days, rectangular currents injected through the recording electrode evoked action potentials. These results demonstrate that unconnected CNS neurons quickly display at least some properties of excitability when cultured in a 3-D ECM. This culture system should facilitate investigation of intrinsic electrical properties of single CNS neurons, and how extrinsic factors including neurotransmitters, hormones, pharmacological agents and contacts with other cells influence electrical activity.

Interactions between the Autonomic Nervous System and Tachycardias in Man

We have endeavoured to present an integrated and dynamic overview of the interaction between the autonomic nervous system and cardiac arrhythmias. Our purpose was not to be comprehensive, detailed, or all inclusive but to present a clinically relevant summary of some of the areas where autonomic tone may be pertinent. Clearly, most arrhythmias are not the result of a primary autonomic nervous system disorder. However, subtle and overt alterations in autonomic tone, interacting with rate, pressure, cardiac volume, contractility, and the intrinsic myocardial electrophysiologic properties likely exert a critical influence on many cardiac arrhythmias. We wish to emphasize that autonomic effects may be temporarily concealed or overwhelmed during certain states or by the administration of drugs. However, the modulating role of neural tone and neurotransmitters is almost always present, contributing significantly to the outcome of therapy.

Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties

1. Reliable methods for establishing fetal mouse spinal cord (SC) and dorsal root ganglion (DRG) cells in long term (greater than 1 mo) dissociated cell cultures are described. These cells have been studied by morphologic and intracellular electrophysiologic techniques. 2. Cells studied electrophysiologically can be relocated after preparation for electron microscopy and examined in thin sections. The electron microscope shows that the surface membranes of these cells were directly accessible to the culture medium. The surfaces of SC cells were studded with synaptic boutons, whereas the DRG cell surfaces generally had none. 3. Current-voltage relationships and linear electrotonic properties of the neurons are described. Delayed and anomalous rectification were seen in both cell types. The length of SC cell dendrites was about one characteristic electrotonic length, while little or no contribution of the relatively sparse DRG cell processes was seen in the transient responses of the DRG cells. 4. Postspike and posttetanic hyperpolarizations in DRG cells were due to a surface membrane conductance increase; this was probably primarily an increase in K+ conductance. Post-activation hyperpolarization in SC cells was primarily due to activation of an electrogenic Na+ pump.

The action of acetylcholine and L-glutamic acid on rat caudate neurons

Acetylcholine plays a crucial role in the regulation of neural functions, including dopamine release, synaptic activity, and intrinsic electrophysiological properties of the nucleus accumbens (NAc) shell. Although the effects of acetylcholine on the action potential properties of NAc medium spiny (MS) neurons have been reported, how intrinsic acetylcholine released from NAc cholinergic interneurons regulates the neural activity of MS neurons is still an open issue. To explore the cholinergic effects on the subthreshold responses and action potential properties of MS neurons in the NAc shell, we first tested the effects of carbachol, a non-selective cholinergic agonist, on MS neuronal activity. Then, we tested the effects of the activation of cholinergic interneurons on the electrophysiological properties of MS neurons via multiple whole-cell patch-clamp recordings. Bath application of carbachol induced resting membrane potential depolarization accompanied by an increase in the voltage response to negative current injection. These increases were blocked by the pre-application of pirenzepine, an M1 muscarinic receptor antagonist. In spite of the facilitative effect on voltage responses of negative current injection, carbachol diminished the characteristic slowly-depolarizing ramp potentials, which respond to positive current pulse injection. Thus, carbachol increased the rheobase and shifted the frequency–current curve toward the right. Repetitive spike firing of a cholinergic interneuron following positive current injection induced a similar increase in the rheobase, which delayed the action potential initiation in 38.9% MS neurons. In contrast to the bath application of carbachol, cholinergic interneuronal stimulation had little effect on the resting membrane potential in MS neurons. These results suggest that the acetylcholine released from a cholinergic interneuron is sufficient to suppress the repetitive spike firing of the adjacent MS neurons, although the depolarization of the resting membrane potential may require simultaneous activation of multiple cholinergic interneurons.