Voltage-gated currents in identified rat olfactory receptor neurons

Whole-cell recording techniques were used to characterize voltage-gated membrane currents in neonatal rat olfactory receptor neurons (ORNs) in cell culture. Mature ORNs were identified in culture by their characteristic bipolar morphology, by retrograde labeling techniques, and by olfactory marker protein (OMP) immunoreactivity. ORNs did not have spontaneous activity, but fired action potentials to depolarizing current pulses. Action potentials were blocked by tetrodotoxin (TTX), which contrasts with the TTX-resistant action potentials in salamander olfactory receptor cells (e.g., Firestein and Werblin, 1987). Prolonged, suprathreshold current pulses evoked only a single action potential; however, repetitive firing up to 35 Hz could be elicited by a series of brief depolarizing pulses. Under voltage clamp, the TTX-sensitive sodium current had activation and inactivation properties similar to other excitable cells. In TTX and 20 mM barium, sustained inward current were evoked by voltage steps positive to -30 mV. This current was blocked by Cd (100 microM) and by nifedipine (IC50 = 368 nM) consistent with L-type calcium channels in other neurons. No T-type calcium current was observed. Voltage steps positive to -20 mV also evoked an outward current that did not inactivate during 100-msec depolarizations. Tail current analysis of this current was consistent with a selective potassium conductance. The outward current was blocked by external tetraethylammonium but was unaffected by Cd or 4-aminopyridine (4-AP) or by removal of external calcium. A transient outward current was not observed. The 3 voltage-dependent conductances in cultured rat ORNs appear to be sufficient for 2 essential functions: action potential generation and transmitter release. As a single odorant-activated channel can trigger an action potential (e.g., Lynch and Barry, 1989), the repetitive firing seen with brief depolarizing pulses suggests that ORNs do not integrate sensory input, but rather act as high-fidelity relays such that each opening of an odorant-activated channel reaches the olfactory bulb glomeruli as an action potential.     

Local stimulation induced GABAergic response in rat striatal slice preparations: Intracellular recordings on QX-314 injected neurons

Gamma-aminobutyric acid (GABA)ergic responses evoked by electrical stimulation in the neostriatal slice preparation were studied in neurons injected intracellularly with Na-conductance blocker QX-314. Local stimulation elicited depolarizing postsynaptic potentials (DPSPs) in the QX-314-injected neurons when the membrane potential was morenegative than −60 mV. When DPSPs were minimized by depolirizing current injection in the QX-314-injected neuron, hyperpolarization was clearly observed following local stimulation. The maximum duration of the hyperpolarizing response to strong local stimulation was about 130 ms. The hyperpolarizing response was blocked by the addition of bicuculine or picrotoxin to the Ringer solution. Intracellular Cl^- injections produced changes in the pattern of the local stimulations-induced responses; the initial depolarizing response was followed by a relatively large amplitude long duration depolarization. The polarity of the long duration of depolarizing response could not be reversed by depolarizing currents which were normally sufficient to reverse the polarity of DPSPs in the neurons without Cl^- injection. The application of pentobarbital enhanced the amplitude and the duration of the hyperpolarizing responses. The revealed potential of the pentobarbital-enhanced response was estimated to be −60 mV. On the basis of their reversal potential, sensitivity to injected Cl^-, sensitivity to GABA blockers picrotoxin and bicuculine, and the effect of pentobarbital, these hyperpolarizing responses are shown to be GABAergic Cl^- mediated inhibitory postsynaptic potentials (IPSPs).     

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.     

Whole-cell recording from honeybee olfactory receptor neurons: ionic currents, membrane excitability and odourant response in developing workerbee and drone

Whole-cell recording techniques were used to characterize ionic membrane currents and odourant responses in honeybee olfactory receptor neurons (ORNs) in primary cell culture. ORNs of workerbee (female) and drone (male) were isolated at an early stage of development before sensory axons connect to their target in the antennal lobe. The results collectively indicate that honeybee ORNs have electrical properties similar, but not necessarily identical to, those currently envisaged for ORNs of other species. Under voltage clamp at least four ionic currents could be distinguished. Inward currents were made of a fast transient, tetrodotoxin-sensitive sodium current. In some ORNs a cadmium-sensitive calcium current was detected. ORNs showed heterogeneity in their outward currents: either outward currents were made of a delayed rectifier type potassium current, which was partially blocked by tetraethyl ammonium or quinidine, or were composed of a delayed rectifier type and a transient calcium-dependent potassium current, which was cadmium-sensitive and abolished by removal of external calcium. The proportion of each of the two outward currents, however, was different within the ORNs of the two sexes suggesting a gender-specific functional heterogeneity. ORNs showed heterogeneity in action potential firing properties: depolarizing current steps elicited either one action potential or, as in most of the cells, it led to repetitive spiking. Action potentials were tetrodotoxin-sensitive suggesting they are carried by sodium. Odourant stimulation with different mixtures and pure substances evoked depolarizing receptor potentials with superimposed action potentials when spike threshold was reached. In summary, honeybee ORNs are remarkably mature at early stages in their development.     

An aminopyridine-sensitive, early outward current recorded in vivo in neurons of the precruciate cortex of cats using single-electrode voltage-clamp techniques

Studies were performed in cortical neurons to determine if voltage- and time-dependent membrane currents could be recognized and characterized in the dynamic, in vivo state. Intracellular measurements made in neurons of the precruciate cortex of awake cats with single-electrode voltage-clamp (SEVC) techniques disclosed an early outward current to depolarizing command steps in 124 of 137 cells studies. The voltage-dependent properties of the early outward current closely resembled those of A-currents studied in vitro in verterbrate and invertebrate neurons. The current was activated rapidly at onset latencies of less than two ms, fell to flat plateau levels within 60–120 ms during sustained depolarization, and was reduced or eliminated in 22 of 23 cells following intracellular administration of 3- or 4-aminopyridine. The magnitude of outward current in response to depolarizing commands was increased by preceding steady hyperpolarization and reduced by preceding steady depolarization. (The steady potentials were of 9.8 s duration and ±40 mV apart from the holding potentials.) Since return to the holding potentials occurred 80 ms before the onset of the command steps, the changes in membrane properties that were induced lasted beyond cessation of the steady polarizing stimuli themselves. Spiking did not prevent recognition of the early outward current as judged from its appearance before and after intracellular application of QX-314 to reduce spike activity. Apart from fast inward currents associated with spike potentials, the early outward current was the most conspicuous and characteristic membrane current noted in these recordings. An additional current component that was noted but not characterized in these studies was a slow, depolarization-induced inward current that could be reduced by intracellular injection of QX-314.     

Corticosterone Reduces Synaptic Inhibition in Rat Hippocampal and Neocortical Neurons in vitro

We examined the effect of corticosterone (10(-7) to 10(-5) M) on membrane properties and postsynaptic potentials, by means of intracellular recordings from neocortical and hippocampal CA1 pyramidal neurons of the intact adult rat in vitro. Corticosterone reduced both the early and the late components of the orthodromically-evoked inhibitory postsynaptic potential in both structures. The glucocorticoid receptor antagonist RU 38486 (10(-6) M) prevented this effect in the hippocampus. In hippocampal, but not in neocortical pyramidal neurons, corticosterone reduced a depolarizing membrane transient evoked by a depolarizing current step and increased the threshold for eliciting action potentials evoked by depolarizing current pulses. Corticosterone did not detectably alter the afterhyperpolarization following repetitive neuronal discharges evoked by current injection, in either the neocortex or in the hippocampus. Excitatory postsynaptic potentials, action potentials, membrane potential and membrane input resistance were also unchanged. The decrease in synaptic inhibition together with the reduction of electrical excitability in the hippocampus, would imply a modulation of response characteristics in pyramidal neurons such that repeated synaptic inputs become more efficient and low frequency input is blunted.     

Electrophysiologic properties of red nucleus neurons in the rat brain slices]

Intracellular recordings from the red nucleus (RN) neurons were made in experiments on the rat brain slices. Passive membrane properties (input resistance and membrane time constant) of the RN neurons were evaluated. Phenomena of potential-dependent rebound depolarization and time-dependent inward rectification were revealed by means of passing hyperpolarizing current pulses through the recorded cells. Injections of depolarizing currents caused repetitive firing of neurons with frequencies directly depending on the intensity of injected currents. Repetitive firing was also characterized by a fast frequency adaptation during injections of currents of different intensities. Stimulation of a region of slices presumably corresponding to the decussation of brachium conjunctivum evoked mainly monosynaptic EPSPs with a "fast"-rise time in the RN neurons, which suggests activation of the synaptic input from the cerebellar nucleus interpositus. Stimulation of the same region sometimes evoked EPSP-IPSP mixtures or pure IPSPs in the RN neurons.     

Electrical membrane properties of rat subthalamic neurons in an in vitro slice preparation

The electrical membrane properties of subthalamic (STH) neurons and their response characteristics to stimulation of the internal capsule (IC) were studied in an in vitro slice preparation. Most STH neurons recorded exhibited spontaneous repetitive firing. The input resistance of STH neurons was 146 +/- 48 M omega and showed both an anomalous and a delayed rectification when the membrane was hyperpolarized or depolarized by current injections. In neurons with the membrane potential less negative than 65 mV, depolarizing current pulses generated repetitive firing with the maximum frequency of up to 500 Hz. Two types of tetrodotoxin (TTX)-resistant cobalt-sensitive potentials, slow depolarizing potential and slow action potential, were observed in STH neurons. The slow depolarizing potential had a long duration (over 500 ms in some cases) and was able to trigger repetitive firing. The slow action potential had a duration of about 30 ms and triggered a burst of firing. The slow action potential was seen only when the neurons were hyperpolarized to more negative than 65 mV by a current injection. Electrical stimulation of IC evoked monosynaptic inhibitory postsynaptic potentials (IPSPs) in most of the neurons examined. The polarity of IPSPs was reversed in the depolarizing direction by intracellular injection of Cl-. Bath application of bicuculline markedly suppressed IPSPs and unmasked monosynaptic excitatory postsynaptic potentials (EPSPs). The EPSP was able to trigger a slow depolarization with repetitive firing or a slow action potential with burst of firing when the neuron was hyperpolarized by a continuous current injection. The results demonstrated that STH neurons in an in vitro preparation have spontaneous discharges, high input resistance, capability to generate high-frequency firing, and Ca potentials. The pattern of responses of STH neurons to synaptic inputs is dependent on their membrane potentials.     

Electrophysiological properties of neuroendocrine cells of the intact rat pars intermedia: multiple calcium currents

Intracellular recordings for current and voltage clamping were obtained from 130 neuroendocrine cells of the pars intermedia (PI) in intact pituitaries maintained in vitro. Spontaneous and evoked action potentials were blocked by TTX or by intracellular injection of a local anesthetic, QX-222. After potassium (K+) currents were blocked by tetraethylammonium (TEA), 4-aminopyridine, and intracellular cesium (Cs+), 2 distinct calcium (Ca2+) spikes were observed which were differentiated by characteristic thresholds, durations, and amplitudes. Both Ca2+ spikes were blocked by cobalt (Co2+) but were unaffected by TTX or QX-222. The low-threshold spike (LTS) had a smaller amplitude and inactivated when membrane potential was depolarized past -40 mV or when evoked at a fast rate (greater than 0.5 Hz). The high-threshold spike (HTS) typically had a larger amplitude and longer duration, was not inactivated at potentials which inactivated the LTS, and could be evoked at rates of up to 10 Hz. Single-electrode voltage-clamp analysis revealed that 3 distinct components of the Ca2+ current were present in most cells. From a negative holding potential (-90 mV), 2 separate peak inward currents were observed; a low-threshold transient current, similar to a T-type Ca2+ current, activated at -40 mV, whereas a large-amplitude inactivating current activated above -20 mV. This large inactivating Ca2+ current was significantly inactivated at a holding potential of -40 mV or by brief prepulses to positive potentials, and was similar to an N-type Ca2+ current. A sustained Ca2+ current (L-type) was observed which was not altered by different holding potentials.(ABSTRACT TRUNCATED AT 250 WORDS)     

Long-term potentiation in the hippocampus using depolarizing current pulses as the conditioning stimulus to single volley synaptic potentials

The conditions responsible for the associative properties of long-term potentiation (LTP) were examined in the CA1 region of the hippocampal slice preparation. Intracellularly recorded EPSPs resulting from single-volley stimulation at low frequency (0.15-0.1 Hz) in the stratum radiatum or oriens were paired with depolarizing current pulses (50-100 msec) injected through the recording microelectrode. It is shown that these EPSPs, when paired with pulses of sufficient magnitude, become potentiated. This potentiation generally reached a peak after 20-30 pairing events and could outlast the conditioning period by more than 1 hr. It was specific to the paired input, was blocked by 2-amino-5-phosphonovalerate (APV) and was largely blocked by prior homosynaptic tetanization (and vice versa). In experiments performed with picrotoxin (PTX) in the bath, EPSPs were potentiated using 2-4 nA current pulses, with somewhat higher values in normal solution. The effective current pulses, in both normal and PTX solution, produced a repetitive spike discharge of 7-11 spikes (per 100 msec), and within this range, higher frequencies were associated with larger potentiations. However, since similar degrees of EPSP potentiation were observed following blockade of spike activity by intracellular QX-314, spike activity was not the primary conditioning factor. For the potentiation to appear, the EPSP had to occur together with the current pulse or precede it by less than about 100 msec. No potentiation was observed when the EPSP immediately succeeded the pulse. The results suggest that the cooperativity aspect of LTP is related to a need for sufficient postsynaptic depolarization.(ABSTRACT TRUNCATED AT 250 WORDS)     

Thalamic bursting mechanism: an inward slow current revealed by membrane hyperpolarization

Hyperpolarization of the ventrolateral thalamic cell membrane reveals a slow inward current which is not normally observed at the resting membrane potential. The response evoked by depolarizing synaptic potentials or depolarizing current pulses from a hyperpolarized potential consists of a burst of action potentials superimposed upon a slow voltage response, in contrast to the single active response evoked without the background polarization. We propose that such behavior is caused by a slow inward current that is activated at subthreshold potentials and inactivated or masked at resting potential.     

Angiotensin II Induces Inward Currents in Subfornical Organ Neurones of Rats

The action of angiotensin II on subfornical organ (SFO) neurones was studied using whole-cell current and voltage-clamp recordings in rat slice preparations. In the current-clamp mode, membrane depolarization in response to angiotensin II was accompanied by an increased frequency of action potentials and an increased membrane conductance. In the voltage-clamp mode, angiotensin II elicited inward currents in a dose-dependent manner. The net angiotensin II-induced inward currents were voltage-independent, with a mean reversal potential of -29.8 +/- 6.2 mV. Amplitudes of the angiotensin II-induced inward currents were decreased during perfusion with a low sodium medium. The angiotensin II-induced inward currents were blocked by the AT1 antagonist losartan, and were partially blocked by the AT2 antagonist PD-123319. Neurones which were sensitive to angiotensin II were found in the peripheral region of the SFO, whereas neurones in the central region were less sensitive to angiotensin II. These results suggest that angiotensin II induces inward currents, with opening of nonselective cation channels through mainly AT1 receptors in a subpopulation of SFO neurones of rats.     

Ca2+ involvement in the action potential generation of myenteric neurones in the rat oesophagus

Intracellular recordings were used to study the physiological behaviour of rat oesophageal myenteric neurones, which are embedded in striated muscle. Injection of depolarizing pulses evoked action potentials with a clear 'shoulder' in all neurones. This shoulder disappeared under low Ca2+/high Mg2+ conditions. Tetrodotoxin (TTX; 1 micromol L-1) did not impede spike firing, whereas under combined TTX and low Ca2+/high Mg2+ conditions the action potentials were completely abolished, indicating that TTX- resistant action potentials are mediated by a Ca2+ current. Further experiments with omega-conotoxin GVIA (100 nmol L-1) revealed that these Ca2+ currents enter the cell via N-type voltage-activated Ca2+ channels (see also accompanying paper). Tetraethylammonium (10 mmol L-1) caused broadening of the action potentials, which probably resulted from prolonged Ca2+ influx due to blockade of the delayed rectifier K+ channel. Although Ca2+ appears to be involved in the spike generation of all rat oesophageal myenteric neurones, only a minority (14%) shows a slow afterhyperpolarization. Thus, no strict correlation exists between the presence of a shoulder and a slow afterhyperpolarization. Furthermore, morphological identification of 25 of the impaled neurones revealed that there was no strict correlation between morphology and electrophysiological behaviour. Consequently, rat oesophageal myenteric neurones appear to differ in several aspects from myenteric neurones in smooth muscle regions of the gastrointestinal tract.     

Multisite optical recording of excitability in the enteric nervous system

A multisite optical recording technique consisting of an array of 464 photodiodes was used to measure dynamic changes in transmembrane potentials (Vm) of guinea-pig and mouse enteric neurones stained with the voltage-sensitive dye Di-8-ANEPPS. Optical recordings of Vm changes in enteric neurones which were evoked by depolarizing current pulses or synaptic activation mirrored the Vm changes measured intracellularly in the same neurone. Action potentials had fractional change in fluorescence of -0.09 +/- 0.06% and their peak to peak noise level was 20 +/- 14% of the action potential amplitude. Optical recordings after electrical stimulation of interganglionic nerve strands revealed slow EPSPs, nicotinergic supra- and subthreshold fast EPSPs as well as propagation of action potentials along interganglionic strands. Local application of acetylcholine onto a single ganglion induced reproducibly and dose dependently action potential discharge demonstrating the feasibility of neuropharmacological studies. The optical mapping made it possible to record action potentials simultaneously in a large number of neurones with high spatiotemporal resolution that is unattainable by conventional techniques. This technique presents a powerful tool to study excitability spread within enteric circuits and to assess differential activation of enteric populations in response to a number of stimuli which modulate neuronal activity directly or through synaptic mechanisms.     

Electrical membrane properties of rat substantia nigra compacta neurons in an in vitro slice preparation

The electrical membrane properties of rat substantia nigra pars compacta (SNC) neurons were studied in an in vitro slice preparation. Some of the recorded neurons were intracellularly labeled with HRP and were found to have morphological characteristics resembling the presumed SNC dopaminergic neurons, as reported by others. The input resistance of SNC neurons at resting membrane potential ranged between 70 and 250 M omega. The membrane resistance showed strong anomalous rectification when the membrane was hyperpolarized by current injection. The anomalous rectification was decreased by the addition of tetraethylammonium bromide (TEA) to the bathing Ringer solution. Injection of depolarizing current or termination of hyperpolarizing current induced slow depolarizing potentials. Their amplitude was dependent on the membrane potential and the current intensity. In neurons treated with tetrodotoxin (TTX) and TEA, slow action potentials were triggered from the slow depolarizing potentials. Both the slow depolarizing potential and slow action potential were TTX resistant and abolished by superfusion of Ca2+-free medium. Long duration hyperpolarizations were observed following the injection of depolarizing current pulses. The hyperpolarization was abolished by the superfusion of Ca2+-free medium or decreased by addition of TEA to the Ringer solution indicating an involvement of a Ca2+-dependent K+-conductance in generation of the hyperpolarization. The long duration hyperpolarization was also observed following action potentials. The spike after hyperpolarization consisted of an initial short duration fast component and a long lasting component. The amplitude of both components seems to be reduced but not abolished by TEA (up to 10 mM). When hyperpolarizing current pulses were applied to neurons that were held either continuously depolarized or were superfused with Ca2+-free medium, the pattern of the membrane potential after the offset of current pulses consisted of an initial fast and a later slow ramp-shaped phase. The latter was associated with a membrane conductance increase and interpreted to be due to an early K+ current. This early K+ current was relatively resistant to TEA. Injections of strong depolarizing currents triggered action potentials with multiple inflections on their rising phase. The amplitudes of action potentials changed abruptly during current application. These data indicate that SNC neurons have multiple generation sites for action potential.     

Intracellular injection of apamin reduces a slow potassium current mediating afterhyperpolarizations and IPSPs in neocortical neurons of cats

Electrophysiologic effects of intracellularly injected apamin, a Ca2+-dependent K+ channel blocker, were investigated in neurons of the motor cortex of awake cats. Single-electrode voltage clamp techniques were used to measure changes in membrane currents including those that were synaptically activated. All changes occurred within 2-4 min after pressure injection of apamin with partial recovery observed within 8-15 min. Apamin selectively abolished an outward current that mediated a slow afterhyperpolarization (AHP) following intracellular depolarizing current pulses and action potentials without influencing the time course of the action potentials or an associated fast AHP component. In addition apamin increased the number and frequency of spike discharges evoked by the depolarizing current pulses and produced a small increase in the rate of background firing activity. The baseline resting potential and input resistance were essentially unchanged by apamin. Apamin also diminished a late, slowly decaying component of inhibitory postsynaptic potentials (IPSPs) and currents (IPSCs) elicited by stimulation of the ventrolateral thalamus or the pyramidal tract. The apamin-induced changes were concomitant with a decrease of the decay time constant of both IPSPs and IPSCs and a positive shift in their reversal potential. The results suggest that the late, slowly decaying component of these inhibitory postsynaptic responses is generated by an apamin-sensitive Ca2+-dependent K+ conductance which is also responsible for the slow AHP.     

Electrophysiology of sipatrigine: a lamotrigine derivative exhibiting neuroprotective effects

Sipatrigine (BW619C89), a derivative of the antiepileptic agent lamotrigine, has potent neuroprotective properties in animal models of cerebral ischemia and head injury. In the present study we investigated the electrophysiological effects of sipatrigine utilizing intracellular current-clamp recordings obtained from striatal spiny neurons in rat corticostriatal slices and whole-cell patch-clamp recordings in isolated striatal neurons. The number of action potentials produced in response to a depolarizing current pulse in the recorded neurons was reduced by sipatrigine (EC(50) 4.5 microM). Although this drug preferentially blocked action potentials in the last part of the depolarizing current pulse, it also decreased the frequency of the first action potentials. Sipatrigine also inhibited tetrodotoxin-sensitive sodium (Na(+)) current recorded from isolated striatal neurons. The EC(50) for this inhibitory action was 7 microM at the holding potential (V(h)) of -65 mV, but 16 microM at V(h) = -105, suggesting a dependence of this pharmacological effect on the membrane potential. Moreover, although the inhibitory action of sipatrigine on Na(+) currents was maximal during high-frequency activation (20 Hz), it could also be detected at low frequencies. The amplitude of excitatory postsynaptic potentials (EPSPs), recorded following stimulation of the corticostriatal pathway, was depressed by sipatrigine (EC(50) 2 microM). This inhibitory action, however, was incomplete; in fact maximal concentrations of this drug reduced EPSP amplitude by only 45%. Sipatrigine produced no increase in paired-pulse facilitation, suggesting that the modulation of a postsynaptic site was the main pharmacological effect of this agent. The inhibition of voltage-dependent Na(+) channels exerted by sipatrigine might account for its depressant effects on both repetitive firing discharge and corticostriatal excitatory transmission. The modulation of Na(+) channels described here, as well as the previously observed inhibition of high-voltage-activated calcium currents, might contribute to the neuroprotective efficacy exerted by this compound in experimental models of in vitro and in vivo ischemia.     

Participation of low-threshold calcium spikes in excitatory synaptic transmission in guinea pig medial frontal cortex

We studied the activation of low-threshold calcium spikes (LTS) by excitatory postsynaptic potentials in pyramidal neurons from guinea pig medial frontal cortex with intracellular recording. We used extracellular bicuculline and phaclofen and intracellular QX-314 to block inhibitory synaptic potentials and sodium currents. Postsynaptic potentials were evoked by stimulation of layer I. We found that large (> 10-15 mV) excitatory synaptic potentials evoked from membrane potentials more negative than -75 mV were able to trigger LTS. The activation of LTS resulted in an increase of the rising slope or amplitude of the synaptic potentials depending on the size of the excitatory postsynaptic potential (EPSP). We used 100 microM NiCl2 to confirm the presence of LTS as part of the EPSPs. The N-methyl-D-aspartate (NMDA) and non-NMDA components of the excitatory synaptic potentials were isolated using (+/-)2-amino-5-phosphonovaleric acid (APV; 50 microM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microM); both components could, independently, trigger an LTS. With recordings made with K+ acetate-filled electrodes, we show that the activation of LTS was critical to allow excitatory synaptic potentials to reach the threshold of action potential firing; also, this amplification of synaptic responses produced the firing of more than a single action potential by the postsynaptic cell. These results demonstrate that in cortical pyramidal neurons the activation of low-threshold calcium spikes results in the amplification of synaptic responses.     

Repetitive action potentials induced in chloride-free solution: Effect of denervation

Isometric mechanical activity and action potentials registered with intracellular microelectrodes were studied in innervated and denervated fibers of the soleus muscle of the rat in normal and chloride-free solutions. The chloride-free solution promoted in both innervated and denervated fibers an increment in the resting membrane potential. The innervated muscles showed long mechanical relaxation and repetitive action potentials after a single depolarizing pulse. On the contrary, denervated muscles were resistant to show mechanical and electrical changes in the chloride-free medium. Spontaneous and evoked action potentials from innervated muscle fibers were abolished by tetrodotoxin. The evoked action potentials generated in denervated fibers had a slower time course and were resistant to tetrodotoxin. After 7 to 10 days of denervation the input resistance was increased by about 30%. Substitution of chloride with sulfate resulted in a 150% increase in input resistance of innervated muscle fibers and 80% in denervated preparations. Alterations in the ionic conductances, a decrease in the maximum rate of rise of the action potentials, and changes in the sodium current kinetics could be the main factors for the absence of repetitive action potentials in denervated fibers exposed to the chloride-free medium.     

Properties of depolarizing plateau potentials in aminopyridine-induced ictal seizure foci of cat motor cortex

The mechanisms of generation of self-sustained depolarizing plateau potentials (DPs) were studied in intracellular recordings in aminopyridine-induced ictal seizure foci in the motor cortex of the cat. In some experiments single-electrode voltage clamp techniques were used and intracellular pressure injection of aminopyridine (Ap), phorbol esters (PhEs) and tetraethylammonium (TEA) was carried out. After several ictal episodes, DPs with bursts of action potentials or with spike inactivation developed gradually in the clonic and interictal phases, without synchronism with surface ictal seizure potentials. In many cases DPs were followed by hyperpolarizing afterpotentials and neuronal inhibition. In bursting neurons DPs originated from the augmented depolarizing envelope of bursts of action potentials. In non-bursting neorons DPs were initiated from summated depolarizing afterpotentials and slow spikes with high threshold, resembling Ca-spikes. In a few neurons DPs were triggered by enlarged excitatory postsynaptic potentials. It was possible to evoke DPs by injections of depolarizing current pulses into single neurons of the Ap-focus, or by intracellular injection of AP, PhEs or TEA. We conclude that DPs are not causal cellular bases of the ictal paroxysmal discharges, rather they occur as consequences of abnormal neuronal activity. It is suggested that DPs are intrinsic regenerative membrane events induced by a transient dominance of voltage-dependent inward currents (carried primarily by calcium ions although sodium ions may contribute) by simultaneous decreases in concurrent outward potassium currents.