Threshold For Action Potential Generation Research Articles

Piracetam-induced changes in the functional activity of neurons as a possible mechanism for the effects of nootropic agents.

Studies were carried out on the effects of piracetam (4-20 mM) on the electrical activity of identified neurons in the isolated central nervous system of the pond snail in conditions of single-electrode intracellular stimulation and recording. Piracetam-induced changes were seen in 60-70% of the neurons studied. Different parameters showed different sensitivities to piracetam: the most frequent changes were in the action potential generation threshold, the slope and shape of the steady-state voltage-current characteristics of neuron membranes, and the appearance of piracetam-induced transmembrane ion currents. Nifedipine and cadmium ions, both of which are calcium channel blockers, generally reversed or weakened the effects of piracetam on the changes seen in test cells. This indicates that the effects of piracetam result from its action on calcium channels; selective changes in calcium channels may determine which piracetam-induced effects appear at the cellular level. It is hypothesized that the piracetam-sensitive cellular plasticity mechanisms may make a significant contribution to its nootropic action at the behavioral level.

Subthreshold response to white-noise current input in a tapering cable model of a neuron

The expectation (mean) and variance of the depolarization in the absence of a threshold for action-potential generation is obtained in a neuron model represented by a tapering equivalent cable with a random (white noise) synaptic input current at a point along the dendrites. The results show that the introduction of a taper in the equivalent-cable representation of the neuron produces larger values for both the expectation and variance which are neither insensitive nor symmetrical with respect to the location of the input. It is also shown that taper extends the invariance of the variability in the steady-state somatic response to proximally located random inputs, implying that only small changes in the noisiness of the somatic response occur for a random input located in the dendrites.

GABAA receptor activation and the excitability of nerve terminals in the rat posterior pituitary.

1. The activation of GABAA receptors in nerve terminal membranes gates a Cl- channel. Experiments were conducted to determine how the activation of this receptor influences membrane potentials, action potentials and voltage-activated Na+ and K+ channels. 2. When activation of the GABAA receptor produced only conductance changes and no voltage changes, action potentials changed only slightly. The threshold for action potential generation increased by 15%. GABA reduced the broadening of action potentials caused by high frequency stimulation by only 7%. These results indicate that membrane shunting by GABA-gated Cl- channels plays a relatively minor role. 3. By recording changes in the current through K+ channels in cell-attached patches, the activation of GABAA receptors was shown to depolarize the nerve terminal membrane from rest by 14 mV. The GABAB receptor agonist baclofen produced no change in resting membrane potential as measured by this same technique. 4. In whole-terminal recordings under current clamp, with pipettes containing various Cl- concentrations, the GABA-induced depolarization increased with Ecl. The variation with Ecl provided a basis for evaluating the contributions of leak and K+ current in the balance of currents that determines the magnitude of the GABA-induced depolarization. 5. Based on the GABA-induced voltage change and an evaluation of the other currents of significance in the relevant voltage range, an estimate was obtained for ECl of -48 mV to give an estimate for the intracellular Cl- ion concentration of 20 mM. 6. Under conditions allowing both conductance and voltage to change during Cl- channel gating, GABA prevented action potential responses to current injection. Comparable depolarizations produced by adjusting a steady holding current also blocked action potential responses. 7. A depolarization from -60 to -45 mV under voltage clamp inactivated approximately 90% of the Na+ channels and activated a small amount of K+ current. This suggests that inactivation of Na+ channels makes a major contribution to the inhibition of action potentials by GABA. 8. These results are consistent with the hypothesis that GABA inhibits neurosecretion by retarding impulse propagation into the terminal arborization. These results support a depolarization block mechanism for the inhibition of secretion, in which depolarization inactivates Na+ channels sufficiently to block action potentials.

A mechanism underlying dopamine D1 and D2 receptor-mediated inhibition of dopaminergic neurones in the ventral tegmental area in vitro.

1. An intracellular recording study was performed to elucidate the mechanism underlying D1 and D2 receptor-mediated inhibition of neuronal activities of dopaminergic neurones in the ventral tegmental area (VTA) using slice preparations of the rat brain. 2. VTA neurones were classified into type I and type II neurones according to the shape of the action potential, which correspond to dopaminergic and non-dopaminergic neurones, respectively. 3. Addition of dopamine (10 microM) and quinpirole (1-100 microM) to the bath hyperpolarized the membrane of the type I neurones concomitantly with an increase in membrane conductance and an inhibition of action potentials which occurred spontaneously and were elicited by depolarizing pulses applied to the cell. However, quinpirole (10 microM) had no effect on the threshold for action potentials induced by a depolarizing pulse. 4. These quinpirole (10 microM)-induced effects were antagonized by simultaneous application of domperidone (5 microM), a D2 receptor antagonist. 5. The amplitude of quinpirole (10 microM)-induced hyperpolarization was decreased by increasing the potassium concentration in the perfusing fluid or simultaneous application of tetraethylammonium (10 microM). 6. SKF 38393 (10 or 100 microM), a D1 receptor agonist, had no effect on the resting membrane potential or action potential firing induced by a depolarizing pulse applied to the cell. However, when SKF 38393 (10 microM) was applied simultaneously with quinpirole (10 microM), the threshold for action potential generation was elevated by 5-6 mV, although there was no enhancement of hyperpolarization induced by quinpirole. 7. The elevation of the threshold for action potentials induced by SKF 38393 in the presence of quinpirole was antagonized by simultaneous application of SCH 23390 (5 microM), a D1 receptor antagonist.8. Dopamine (10 microM), quinpirole (10 or 100 microM) and SKF 38393 (10 or 100 microM) had no effect on the resting membrane potential or spontaneously occurring action potentials in type II neurones.9. These findings suggest that activation of dopamine D2 receptors of dopaminergic neurones in the VTA increases potassium conductance, thereby hyperpolarizing the membrane and eventually inhibiting neuronal activities. They also suggest that simultaneous activation of both D1 and D2 receptors enhances the D2 receptor-mediated inhibitory effects by elevation of the threshold for action potential generation.

Proctolin activates an inward current whose voltage dependence is modified by extracellular Ca2+

The pentapeptide proctolin modulates the activity of the rhythmic pattern generators in the crustacean stomatogastric nervous system. Proctolin strongly excites the lateral pyloric and the inferior cardiac neurons of the stomatogastric ganglion (STG), causing them to fire extended high-frequency bursts of action potentials (Hooper and Marder, 1987; Nusbaum and Marder, 1989a,b). We now report that proctolin depolarizes these cells maximally at membrane potentials close to the threshold for action potential generation. In voltage clamp, proctolin evokes an inward current, carried at least partially by Na+, that shows strong outward rectification. Removal of extracellular Ca2+ markedly increases the amplitude of the proctolin-evoked current and linearizes its current-voltage curve. The properties of the proctolin current make it ideally suited to contribute to the activity-dependent modulation of the pyloric network of the STG.

Dopamine D2 receptor activation depolarizes rat supraoptic neurones in hypothalamic explants.

1. Intracellular current and voltage clamp recordings were obtained from rat supraoptic nucleus neurones in superfused hypothalamic explants in order to evaluate their response to dopamine and to D1 and D2 agonists. 2. With one exception, exposure to dopamine (10-200 microM) depolarized supraoptic neurones. When tested for an effect on twenty-one spontaneously active supraoptic neurones, dopamine enhanced the firing of all eleven continuous-firing (possibly oxytocin-secreting) neurones and prolonged the burst in all ten phasic-firing (vasopressin-secreting) neurones. 3. In sixty-seven of sixty-eight neurones where current injection was used to maintain membrane potential below threshold for action potential generation, current clamp data revealed that exposure to dopamine (10-200 microM) was followed in 10-17 s by a gradual 3-7 mV membrane depolarization that lasted for 4-15 min and was accompanied by a 12-23% reduction in input resistance. Exposure to quinpirole, a D2 agonist (10-200 microM), induced a similar response with comparable onset, duration and change in input resistance. In contrast, tests on sixteen cells indicated little or no response to a D1 agonist SKF38393. 4. Under voltage clamp, dopamine was noted to induce an inward current, accompanied by a 7.5-40% increase in membrane conductance over the corresponding time course. 5. Voltage-current plots for dopamine-induced depolarizations were linear in the range -50 to -110 mV. Dopamine and quinpirole depolarizations had extrapolated mean reversal potentials of -25 +/- 10 mV (mean +/- S.D.) and -20 +/- 15 mV respectively. This approximated the mean reversal potential of -20 +/- 8 mV measured from the dopamine-induced inward current using single-electrode voltage clamp. 6. The actions of dopamine were selectively antagonized by two D2 receptor antagonists, sulpiride and spiperone, but neither influenced membrane depolarizations induced by equimolar concentrations of noradrenaline. Dopamine-induced depolarizations also persisted following selective blockade of alpha 1-adrenergic receptors by prazosin; under these conditions, noradrenaline induced membrane hyperpolarization. 7. Following complete substitution of external Na+ with Tris, the reversal potential for the dopamine-induced response was shifted to -70 +/- 9.8 mV. This value was consistently less negative than the estimated potassium equilibrium potential. 8. The depolarization action of dopamine persisted in media containing tetrodotoxin and with an external calcium concentration ([Ca2+]o) of 0 mM-Ca2+ with 6 mM-Mg2+ or Mn2+, but was abolished following intracellular injection of [1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), a Ca2+ chelator.(ABSTRACT TRUNCATED AT 400 WORDS)

Dentate hilar cells with dendrites in the molecular layer have lower thresholds for synaptic activation by perforant path than granule cells

Neurons in the dentate hilus or area CA3c of rat hippocampal slices were recorded intracellularly with electrodes containing the fluorescent dye Lucifer yellow. Stimulation of perforant path fibers in the molecular layer of the fascia dentata strongly excited most hilar neurons, with a much lower threshold for action potential generation than granule cells and area CA3c pyramidal cells that were recorded in the same area of the slice. Examination of dye-filled hilar neurons with a confocal microscope showed that hilar cells with a low threshold were morphologically heterogeneous: some were spiny "mossy" cells, and others were aspiny interneurons. However, all hilar cells with low thresholds possessed dendrites that penetrated the granule cell layer and passed into the molecular layer, often reaching the outer molecular layer. The few hilar cells that had a threshold similar to, or greater than, granule cells did not possess visible dendrites in the molecular layer. The results suggest that the circuitry of the dentate region allows for (1) excitation of both granule cells and hilar cells by perforant path stimuli, and (2) strong excitation of most hilar cells when most granule cells are subthreshold for action potential generation. Given that hilar neurons project to many different sites in the ipsilateral and contralateral fascia dentata (Blackstad, 1956; Zimmer, 1971; Swanson et al., 1978; Laurberg and Sørensen, 1981), it is quite likely that hilar neurons are involved in the processing of information passing from entorhinal cortex to hippocampus.

Effects of axotomy on synaptic transmission and structure in frog sympathetic ganglia.

Axotomy was carried out on frog sympathetic neurons of the two last lumbar chain ganglia. At different times thereafter, synaptic transmission was analysed electrophysiologically by intracellular microelectrodes and compared with synaptic density, measured by electron microscopy in the same ganglia. For this purpose, modifications in synaptic transmission were estimated first, by the numbers of B and C sympathetic neurons exhibiting subthreshold excitatory postsynaptic potentials in response to 10 Hz orthodromic stimulation of preganglionic fibres, and second, by the amplitude and number of excitatory postsynaptic potentials occurring over 5-10 s periods of 10 Hz stimulation. By distinguishing two types of morphological relationships between the pre- and postsynaptic elements, two contact indices were defined: a synaptic index (ratio of the number of synapses encountered to the number of perikarya explored) and a simple contact index corresponding to the same type of contacts, but without any membrane differentiation. Both the electrophysiological and morphological results showed that the first effects were detectable 4 days after axotomy, and that the main alterations in synaptic transmission and density occurred at 2 weeks. In addition, while in normal ganglia the excitatory postsynaptic potentials of B and C neurons reached the threshold for action potential generation in response to 10 Hz stimulation, about 29% of the axotomized neurons had subthreshold excitatory postsynaptic potentials 1 week after section. At 2 weeks, this proportion reached 65%, and the synaptic and simple contact indices, at 90% and 60% respectively, were significantly lower than the control ganglion indices. At longer times after axotomy, there was a discrepancy between the morphological and electrophysiological results: at 1 month, the synaptic index seemed to rise as the decline in the efficacy of synaptic transmission became more marked. The amplitude of the subthreshold excitatory postsynaptic potentials recorded in B neurons was 5.5 +/- 2.8 mV (mean +/- SD, n = 18); this value was significantly lower by about 50% than that measured 1 week after axotomy. In addition, the number of excitatory postsynaptic potentials in B neurons reached an average maximum of 83 +/- 29 for 100 stimuli applied at 10 Hz. Similar results were obtained for C neurons. Two months after axotomy, the physiological and morphological parameters of synaptic efficacy began to recover and return to normal values, but had not reached them by 4 months. These observations show that some synaptic transmission remains possible, even with a much reduced number of synaptic complexes. It is suggested that after axotomy, simple contacts also might be involved in synaptic transmission.

A reevaluation of excitatory amino acid-mediated synaptic transmission in rat dentate gyrus

1. Intracellular recordings were made from granule cells in combined slices of the hippocampus and parahippocampal cortex from adult Wistar rats. The neurons had a mean resting membrane potential (EM) of -85.1 +/- 0.7 (SE) mV, input resistance (Rin) of 30.9 +/- 1.5 M omega and action potential (AP) amplitude of 79.9 +/- 1.06 mV measured from the threshold potential. The threshold for AP generation was approximately 40 mV positive to EM. 2. The passive current-voltage relationship showed anomalous rectification, with Rin increasing by 34% on average at modest depolarizations. With large excursions of the EM (by +/- 30 mV or more), there was a marked fall in Rin. 3. With extracellular recording, a monophasic, positive-going field potential of 5-15 mV was evoked by stimulation of the perforant path (PP). Intracellularly, an excitatory postsynaptic potential (EPSP) of up to 40 mV in size was recorded. It was unusual to evoke an AP on orthodromic stimulation. Perfusion with picrotoxin (PTX, up to 20 microM) had small and variable effects on the EPSP, which implies that GABAergic inhibition does not play a major role. 4. Tonic depolarization reduced the EPSP. Hyperpolarization either had no effect or again decreased the EPSP. 5. The role of excitatory amino acid (EAA) receptor subtypes in mediation of the EPSP was investigated. Perfusion with the non-N-methyl-D-aspartate (NMDA) receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) caused a dose-dependent reduction of the EPSP with a shift to the right of the input-output relationship. The ED50 for this effect was approximately 2 microM. 6. In the presence of 5-10 microM CNQX a small component of the EPSP usually remained, which could be blocked by the NMDA receptor antagonist +/- 2-amino-5-phosphonovaleric acid (APV, 20-50 microM). This depolarizing component was markedly enhanced during perfusion with Mg2(+)-free medium. It increased in size and duration when the membrane was depolarized and decreased with hyperpolarization. These properties are consistent with the mediation of this potential via NMDA receptors. 7. These results indicate that NMDA receptors contribute to transmission at the synapse between the PP and the granule cell. This was confirmed by demonstrating that APV caused a small reduction in the size of the untreated EPSP and a shortening of the recovery phase.(ABSTRACT TRUNCATED AT 400 WORDS)

GABAA-receptor-activated current in dorsal root ganglion neurons freshly isolated from adult rats

1. gamma-Aminobutyric acid (GABA) is the major inhibitory neurotransmitter affecting dorsal root ganglion (DRG) neurons. This study compares properties of current activated by the GABAA receptor in two populations of DRG neurons. DRG neurons were isolated from adult rat with the use of enzymatic and mechanical means. Within hours of being isolated, neurons were recorded from with the use of the whole-cell variant of the patch-clamp technique. 2. One population of neurons exhibited an afterdepolarizing potential (ADP), a low threshold for action-potential generation (-45 to -50 mV), a short-duration action potential (less than 2 ms) that was abolished in the presence of 1-2 microM tetrodotoxin (TTX), and an insensitivity to 50 nM capsaicin. The second population of neurons exhibited a high threshold for action-potential generation (less than -40 mV), a shoulder on the falling phase of the action potential, insensitivity of action-potential generation to TTX (1-2 microM), and a depolarizing response to application of 50 nM capsaicin. 3. Sensitivity to GABA (over the range of 1-1,000 microM) was comparable for the two populations of neurons. 4. GABA-activated current was greater in ADP neurons than in non-ADP-type neurons of a comparable diameter (30-50 microns). The mean +/- SE amplitude of current activated by 10 microM GABA in ADP neurons was 0.310 +/- 0.050 nA (range = 0.110-0.460 nA, n = 8), and 0.037 +/- 0.016 nA (range = 0.010-0.130 pA, n = 7) in non-ADP neurons. Ten microM GABA elicited cell firing in ADP neurons but not in non-ADP neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibitory modulation of neuronal voltage-dependent sodium current by Phe-Met-Arg-Phe-amide

The tetrodotoxin (TTX)-sensitive, voltage-gated Na +-current ( I Na) in a cluster of peptidergic neurons, involved in egg laying, in the CNS of the mollusc Lymnaea stagnalis, is modulated by the neuropeptide FMRFa (Phe-Met-Arg-Phe-NH 2). Application of FMRFa reversibly reduced the isolated I Na in a dose-dependent fashion. The physiological consequence is that the threshold for action potential generation is increased, causing an arrest of ongoing firing activity. The inhibitory action of FMRFa reported here is the first known example of modulation of the voltage-gated I Na by a putative neurotransmitter in intact nerve cells. This finding underlines the importance of modulation of ionic currents as a mechanism of regulation of neuronal excitability and includes the voltage dependent Na current in the range of currents subject to transmitter modulation.

Action of vasopressin analog on neuronal excitability in Helix lucorum.

1. The vaospressin analog in most cases increased the excitability of command neurons of the defensive reflex inHelix lucorum. 2. This effect is partially reversihle (it is abolished by rinsing out of the substance), but at least in the first stages, it is also cumulative on repeated applications of desglycine-arginine-vasopressin. 3. The mechanism of this action of desglycine-arginine-vasopressin may consist of activation of a pacemaker mechanism and also of a change in the threshold of action potential generation.

Inhibitory control of local excitatory circuits in the guinea-pig hippocampus.

1. Exposure to the gamma-aminobutyric acid antagonist, picrotoxin, causes the discharge of hippocampal pyramidal cells to become synchronized. Synaptic mechanisms underlying the development of synchrony were investigated by recording from pairs of cells in the CA3 region of guinea-pig hippocampal slices. 2. Picrotoxin suppressed unitary inhibitory synaptic events. It appeared not to affect monosynaptic excitatory connections. Picrotoxin revealed latent excitatory connections in seven out of twenty-one dual recordings from burst-firing cells. 3. Post-synaptic events revealed by picrotoxin were elicited rarely by single action potentials. They were evoked with mean latencies of at least 8 ms and with more than 30% failures of transmission by bursts of three or more action potentials. They were suppressed by increasing extracellular Ca2+. They were considered to be mediated by polysynaptic excitatory pathways. 4. Polysynaptic excitatory post-synaptic potentials (e.p.s.p.s) had a smooth rising phase with time-to-peak of 15-40 ms and a falling phase of similar duration. Their amplitude was 2-3 mV at membrane potentials close to -70 mV. This shape was similar to that of summed e.p.s.p.s evoked by a burst of three to six action potentials at monosynaptic connections between CA3 cells. 5. One cell could evoke excitatory synaptic events in more than one follower cell, suggesting that axon collaterals mediating recurrent excitation were divergent. More than one polysynaptic excitatory pathway could exist between two cells. 6. We examined the role of recurrent excitatory synapses in the development of synchrony. As inhibition was suppressed by picrotoxin, simultaneous excitatory synaptic events appeared in recordings from pairs of cells. They occurred rhythmically at intervals of 0.5-3 s and grew in amplitude with time. Synchronous neuronal discharges were observed when the threshold for action potential generation was exceeded. 7. Firing induced in one cell could sometimes evoke a sequence of post-synaptic events in another cell as inhibition was suppressed. Initially, no connection was detected. On adding picrotoxin, a polysynaptic e.p.s.p. was revealed and with time longer-latency components were recruited to the synaptic event. The amplitude of later components grew until firing threshold was reached. 8. We suggest that synchronous firing develops due to the loss of inhibitory control over the spread of firing between CA3 pyramidal cells via divergent, polysynaptic, recurrent excitatory pathways.

Electrophysiological studies in cultured mouse CNS neurones of the actions of an agonist and an inverse agonist at the benzodiazepine receptor.

The action of agents which bind with the benzodiazepine (BZ) receptor has been investigated by use of intracellular recordings from dissociated mouse neurones grown in tissue culture. The agents tested were midazolam (an agonist at the BZ receptor) and methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM-an inverse agonist at the BZ receptor). These were applied to the neurone under study by one of the following methods: iontophoresis; pressure application of known concentrations from blunt pipettes; directly in the perfusing medium. On only very few occasions did midazolam or DMCM have a direct effect on the membrane potential (EM) or conductance (GM) of the impaled neurone. For the neurones where direct effects were present, there was no consistent pattern of response. Neither substance affected the threshold for action potential generation. The effect of midazolam and DMCM on responses evoked by iontophoretic application of gamma-aminobutyric acid (GABA) was also investigated. Three parameters were used to quantify GABA responses: the depolarization (VGABA); the increase in GM (gGABA) measured with constant current pulses; using voltage clamp, the GABA current (IGABA). The GABA response should be quantified by a parameter which is linearly related to the number of GABA-operated channels which are conducting at any instant. VGABA does not fulfil this criterion. gGABA is an appropriate parameter, but is difficult to determine for large responses where the membrane is nearly short circuited. IGABA measured during voltage clamp fulfils this criterion. Midazolam (greater than 10(-6) M) reliably potentiated GABA responses with a parallel shift to the left of the dose-response curve. This is in agreement with biochemical studies where BZs increase the affinity of the GABA receptor for its ligand. The effect of DMCM on GABA responses was more variable. In the majority of cases GABA responses were reduced by DMCM. The threshold dose for this depression was usually around 10(-6) M, but was sometimes as low as 10(-8) M. Dose-response curves of IGABA or gGABA showed the inhibition to be of a non-competitive nature. The maximum inhibition achieved was around 70%. For a given neurone, and at doses which did not necessarily cause a reduction of the response to GABA, DMCM could antagonize the potentiating action of midazolam on GABA responses. A possible interpretation is that more than one BZ site per receptor complex must be occupied by a BZ agonist (or inverse agonist) before the functional changes for the complex as a whole can occur. Desensitization to GABA was increased by midazolam.

Chlormethiazole‐mode of action

Studies in mice demonstrated that the anticonvulsant profile of chlormethiazole differs from that of diazepam and the barbiturates. Chlormethiazole protects animals from convulsions induced by a wide variety of chemoconvulsants known to block the action of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), such as bicuculline, picrotoxin, isoniazid and pentetrazol, thus confirming and extending earlier studies on its broad anticonvulsant characteristics. Chlormethiazole is particularly potent against isoniazid-induced convulsions, which are probably induced by reductions of GABA levels in the brain. Chlormethiazole was found to have a weak action on benzodiazepine receptor binding, GABA receptor binding and kainic acid receptor binding. Chlormethiazole inhibited picrotoxin binding at very high concentrations, but lowered the functional effects of picrotoxin at much lower concentrations than those affecting picrotoxin binding. Moreover, chlormethiazole failed to change GABA or glutamate levels in the brain and did not affect glutamic acid decarboxylase (GAD) activities in the rat brain. Muscimol (a GABAA agonist) enhanced the anticonvulsant activity of chlormethiazole against picrotoxin but not against bicuculline-induced convulsions. Muscimol enhanced the anticonvulsant potency of diazepam against both chemoconvulsants. These data suggest that the anticonvulsant activity of chlormethiazole is not mediated directly through changes in GABA or glutamate levels or by a direct (agonist) action at the GABA or benzodiazepine receptor complex. These findings suggest that chlormethiazole may enhance GABA transmission beyond the GABA receptors, hypothetically at the level of the GABA receptor coupled ionophore (e.g. the chloride ion channel). Applied micro-iontophoretically, chlormethiazole was found to potentiate the inhibitory responses to GABA, muscimol and glycine, but not to acetylcholine. The potentiation of glycine-mediated inhibition is unique for chlormethiazole and does not occur with any other known anticonvulsant (barbiturates, benzodiazepine, phenytoin or sodium valproate). Studies in primary cultures, derived from spinal cord neurones, showed that chlormethiazole produces hyperpolarization together with an increase in the threshold for action potential generation. Further in vitro studies indicated that chlormethiazole acts on some types of Ca2+-dependent chloride ion channels.(ABSTRACT TRUNCATED AT 400 WORDS)

Effect of a volatile anesthetic upon presynaptic excitability in mammalian hippocampus.

Changes in presynaptic terminal axon excitability produced by enflurane in the rat hippocampal slice preparation were investigated by stimulation of Schaeffer collateral terminal axons and by recording single unit antidromic action potentials. Stimulating pulses were preceded by conditioning hyperpolarizing or depolarizing current pulses. A plot of net threshold for action potential generation against the conditioning pulse yields an "accommodation curve;" changes in this curve can be used to assess the mechanism by which changes in excitability are produced. Enflurane, at a concentration equivalent to approximately equal to 1.3 times the minimum alveolar concentration, reduced excitability of terminal axons and increased accommodation in a manner consistent with a possible change in the inactivation of gNa.

Membrane properties of rat locus coeruleus neurones

Intracellular recordings were made from neurones in the locus coeruleus contained within a slice cut from rat pons and maintained in vitro. Most neurones fired action potentials spontaneously at frequencies of between 1 and 5 Hz; this did not arise from spontaneous synaptic input but appeared to result from endogenous properties of the membrane conductances. Under voltage clamp at potentials near threshold for action potential generation (− 55 mV) there was a persistent inward calcium current. This current became less with membrane hyperpolarization and was abolished at about −70 mV. Two potassium currents were observed. The first had properties similar to that generally described as the “fast” potassium current ( I K,A ); it flowed transiently (for about 200 ms) when the membrane potential passed from about −65 to −45 mV, and was blocked by 4-aminopyridine. The second was a calcium-activated potassium current ( I K,Ca ); it flowed for several seconds following a burst of calcium action potentials. Spontaneous and evoked action potentials had both tetrodotoxin-sensitive and tetrodotoxin-resistant components. The latter was apparently due to calcium entry. The potential changes occurring during the spontaneous firing of locus coeruleus neurones could be reconstructed qualitatively from the ionic conductances observed. The membrane properties of the locus coeruleus neurones were remarkably uniform; however, about 5% of cells impaled within the region of the locus coeruleus were electrophysiologically distinct. These atypical cells had short duration action potentials, did not fire spontaneously and had large spontaneous depolarizing synaptic potentials.

Cellular mechanisms for modulation of posture by octopamine and serotonin in the lobster

Injection of the monoamines octopamine and serotonin into the hemolymph of lobsters causes them to assume opposing static postures: octopamine causes a tonic extension of all extremities, while serotonin induces a tonic flexion. These amines have been shown previously to act as circulating neurohormones and to cause contraction of isolated exoskeletal muscles. The opposing postures elicited by amine injection do not appear to arise from peripheral actions, however, since serotonin and octopamine have similar effects on both flexor and extensor muscles. Instead, the amines are present within the central nervous system where they are probably released synaptically to activate coordinated patterns of tonic motor activity generating the observed postures. These motor patterns have been recorded from abdominal ganglia and involve the synchronized activation and inhibition of multiple excitatory and inhibitory motoneurons innervating the postural flexor and extensor muscles. These effects can be observed in an isolated abdominal ganglion. To begin a search for the cellular targets of amine action within the central nervous system, we recorded from identified postural motoneurons in abdominal ganglia. In two motoneurons that act to produce postural extension, octopamine reduced and serotonin raised the apparent threshold for action potential generation recorded in the somata. The amines do not significantly affect the resting potential or input resistance of the motoneurons measured in the cell bodies. Although the amine effect on motoneuron excitability could be due to actions directly on the motoneurons, our evidence suggests that at least part of the effect results from activation or inhibition of premotor interneurons: (1) an increase in EPSP frequency is observed in the motoneurons during octopamine superfusion, while a small decrease is observed during serotonin superfusion; (2) the amine effects on action potential threshold are abolished by cobalt ion. These results suggest that octopamine and serotonin have dual effects on posture: within the central nervous system, they act specifically to generate coordinated motor patterns for flexion or extension; as circulating neurohormones, they act nonspecifically to enhance the responses of exoskeletal muscles to motoneuronal activation.

Effect of substance P on non-cholinergic fast and slow post-stimulus depolarization in the guinea-pig ileum

Membrane potentials were recorded with intracellular electrodes from the circular muscle cells of the guinea-pig ileum, in vitro, in the presence of atropine (1.4 × 10 −6 M) at 30°C. Under such conditions, the preparations did not show any spontaneous activity. Exposure of the tissue to substance P (10 −7−10 −6 M) for a short period of time ( < 5 min) caused depolarization and action potentials. At this time the membrane resistance appeared to be decreased and after transmural nerve stimulation the fast post-stimulus depolarization, which followed the inhibitory junction potential (IJP), triggered action potentials. After prolonged exposure of the tissue to substance P (10 min), the contractile activity of the preparation subsided, the membrane potential was decreased by 6.4 ± 1.1 mV and the membrane resistance was increased; following transmural nerve stimulation, the amplitude of the IJP was increased, the fast post-stimulus depolarization was abolished and the later slow post-stimulus depolarization was enhanced and in some preparations reached threshold for action potential generation. In the latter preparations in the absence of transmural nerve stimulation, the membrane potential occasionally showed regular oscillations of approximately 10 mV in amplitude every 5–8 s. When the tissue was hyperpolarized by anodal current, after prolonged exposure to substance P, the fast post-stimulus depolarization did not return indicating that its abolition was not a secondary effect due to the depolarization of the tissue by substance P.

Plasticity of the receptive field of a molluscan polyfunctional neuron.

1. The structure of the synaptic response of the LPa3 neuron to a single stimulation of any point on the mantle consists of a two-component SPSP. The multicomponent SPSP composition is due to the presence of parallel channels. A signal from one point of the mantle reaches the LPa3 neuron via two independent paths. 2. The receptive field of LPa3 connected with the mantle is nonuniform in its plastic properties. From the same point on the mantle a signal can be transmitted in parallel along a channel of high plasticity and a channel of low plasticity. 3. Simultaneous activation of several parallel channels ensures that the threshold of action potential generation be attained.