Biomedical subjects

L S Benardo

Publications and source records attributed to L S Benardo.

At least 19 recordsLinked to original sources

PubMed2002

Laminar properties of 4-aminopyridine-induced synchronous network activities in rat neocortex.

We examined the effects of 4-aminopyridine (4-AP) on isolated horizontal (superficial, middle and deep) rat neocortical slices in order to study laminar synchronous network behavior directly. Application of 4-AP induced spontaneous synchronized activity in all of these types of slices. In middle and deep layer slices the activities were similar to those of coronal slices, consisting of periodic short- and long-duration discharges. In superficial slices distinct spontaneous rhythmic multiphasic burst discharges were induced. Ionotropic glutamate receptor antagonists blocked the 4-AP-induced synchronous activities in middle and deep layer slices, but those in superficial slices persisted. The GABA(A) receptor antagonist picrotoxin suppressed this spontaneous synchronous activity resistant to 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (a NMDA receptor antagonist) and 6-cyano-7-nitroquinoxaline-2,3-dione (a non-NMDA receptor antagonist), in superficial slices, leaving small, slow spontaneous events. In superficial slices with intact excitatory amino acid transmission, picrotoxin attenuated the 4-AP-induced spontaneous synchronous discharges, even in this highly convulsant environment. By contrast, conventional coronal slices showed robust spontaneous epileptiform discharges under these circumstances. In intact coronal slices focal 4-AP application in superficial layers induced spontaneous inhibitory GABAergic events, while delivery into deep layers led to epileptiform discharges. From these results we conclude that: (1) 4-AP-induced population discharges are driven by glutamatergic transmission in middle and deep layer horizontal slices, and by GABAergic transmission in superficial layers; (2) only superficial layers are capable of supporting synchronized GABAergic activity independent of excitatory amino acid transmission; (3) superficial layers do not sustain epileptiform activity in the absence of deep layer neurons; and (4) synchronized superficial networks can inhibit deep layer neuronal activity.

4-Aminopyridine↗

PubMed2000

Valproate prevents epileptiform activity after trauma in an in vitro model in neocortical slices.

PURPOSE: Epileptogenesis is a hallmark of severe cortical trauma, with up to approximately 80% of patients experiencing seizures in the first 24 hours after penetrating head injury. An in vitro model of traumatic brain injury was developed to investigate hyperexcitability and epileptogenesis and their prevention. We determined whether sodium valproate would prevent epileptiform activity in this experimental model. METHODS: Rat cortical slices were prepared and maintained in vitro using standard methods. Trauma was effected by removing the superficial 450-500 microm of slices. Traumatized slices were exposed to valproate at various time points. Intracellular and extracellular recordings were used to assess evoked activities. RESULTS: In untreated traumatized deep segments, hyperexcitability was manifested by depressed inhibition and often (54%) by epileptiform activity. Preparations exposed to valproate at 30 minutes or later after trauma showed abnormal activity similar to control traumatized slices. Epileptogenicity in deep segments was significantly reduced when slices were exposed to valproate (a) continuously immediately after trauma, (b) after a 20-minute delay, or (c) immediately after trauma for 1 hour and then returned to physiological solution. Finally, slices that were exposed to valproate or pentobarbital beginning 20 minutes after trauma for only 1 hour and then returned to physiological medium showed a significant reduction in abnormal activity. Valproate was found to enhance fast gamma-aminobutyric acid(A)-ergic inhibitory strength. CONCLUSIONS: Valproate significantly reduces epileptiform activity after trauma to the neocortex, likely by restoring the excitation-inhibition balance, perhaps through augmentation of gamma-aminobutyric acid transmission. The timing of this action may have implications for mechanisms of seizure genesis and may suggest a role for rapid treatment.

Animals↗

PubMed1999

Restrictions on inhibitory circuits contribute to limited recruitment of fast inhibition in rat neocortical pyramidal cells.

To further define the operational boundaries on fast inhibition in neocortex, whole cell recordings were made from layer V pyramidal neurons in neocortical slices to evaluate evoked inhibitory postsynaptic currents (IPSCs) and spontaneous miniature IPSCs (mIPSCs). Stimulating electrodes were placed in layers VI and I/II to determine whether simultaneous stimulation of deep and superficial laminae could extend the magnitude of maximal IPSCs evoked by deep-layer stimulation alone. The addition of superficial-layer stimulation did not increase maximal IPSC amplitude, confirming the strict limit on fast inhibition. Spontaneous miniature IPSCs were recorded in the presence of tetrodotoxin. The frequency of spontaneous mIPSCs ranged from 10.0 to 33.1 Hz. mIPSC amplitude varied considerably, with a range of 5. 0-128.2 pA and a mean value of 20.7+/-4.1 pA (n = 12 cells). The decay phase of miniature IPSCs was best fit by a single exponential, similar to evoked IPSCs. The mean time constant of decay was 6.4+/-0.6 ms, with a range of 0.2-20.1 ms. The mean 10-90% rise time was 1.9+/-0.2 ms, ranging from 0.2 to 6.3 ms. Evaluation of mIPSC kinetics revealed no evidence of dendritic filtering. Amplitude histograms of mIPSCs exhibited skewed distributions with several discernable peaks that, when fit with Gaussian curves, appeared to be spaced equidistantly, suggesting that mIPSC amplitudes varied quantally. The mean separation of Gaussian peaks ranged from 6.1 to 7.8 pA. The quantal distributions did not appear to be artifacts of noise. Exposure to saline containing low Ca(2+) and high Mg(2+) concentrations reduced the number of histogram peaks, but did not affect the quantal size. Mean mIPSC amplitude and quantal size varied with cell holding potential in a near-linear manner. Statistical evaluation of amplitude histograms verified the multimodality of mIPSC amplitude distributions and corroborated the equidistant spacing of peaks. Comparison of mIPSC values with published data from single GABA channel recordings suggests that the mean mIPSC conductance corresponds to the activation of 10-20 GABA(A) receptor channels, and that the release of a single inhibitory quantum opens 3-6 channels. Further comparison of mIPSCs with evoked inhibitory events suggests that a single interneuron may form, on average, 4-12 functional synapses with a pyramidal cell, and that 10-12 individual interneurons are engaged during recruitment of maximal population IPSCs. This suggests that inhibitory circuits are much more restricted in both the size of the unit events and effective number of connections when compared with excitatory inputs.

Animals↗

PubMed1998

Removal of superficial inhibition releases hyperexcitability in middle and deep horizontal slices from rat somatosensory neocortex.

Synaptic physiology was studied in horizontal slices of rat somatosensory neocortex. Intrinsic properties of pyramidal neurons from horizontal slices resembled those recorded in cells from coronal slices, but cells in superficial horizontal slices displayed more prominent fast and slow inhibition, while cells from deeper slices showed disinhibition. This disinhibition in deeper horizontal slices resulted in epileptogenesis in 81% of middle and 35% of deep layer horizontal slices. Brief exposure to glutamate antagonists and dantrolene was ineffective in preventing epileptic activity, but limited pentobarbital exposure reduced the proportion of deep slices manifesting epileptiform activity by 75%. Thus, within cortex inhibition dominates superficially and excitation predominates in deeper layers. While the cortex is vulnerable to hyperexcitability when superficial cortex is compromised, enhancing fast inhibition can reset the excitation-inhibition balance, and prevent epileptogenesis.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1998

Synchronous firing of inhibitory interneurons results in saturation of fast GABA(A) IPSC magnitude but not saturation of fast inhibitory efficacy in rat neocortical pyramidal cells.

The kinetic properties of evoked fast inhibitory postsynaptic currents were examined to elucidate factors underlying the limit on the magnitude of fast inhibition in neocortex. Using whole-cell voltage-clamp recordings from layer V pyramidal neurons in slices of rat somatosensory cortex, fast gamma-aminobutyric acid-A (GABA[A])ergic inhibitory postsynaptic currents were selectively recorded by holding cells at potentials equal to excitatory postsynaptic current reversal (approximately 0 mV). As stimulus intensity was increased, the magnitude and duration of the fast inhibitory postsynaptic current increased. Over the range of stimuli applied (2-10 V), fast GABA(A)-mediated inhibitory postsynaptic currents reached a maximum peak conductance of 25.9 +/- 4.2 nS (range 10.5-41.2 nS) at intensities approximately 2-times threshold. As stimulus intensities were increased beyond this point of maximal conductance, the time constant of current decay increased as function of stimulus strength, while rise time remained unaffected. Exposure to nominally magnesium-free solutions did not affect maximal peak conductances of fast inhibitory postsynaptic currents, but did cause an increase in the time constants of current decay by 66.3 +/- 23.6%, resulting in an 85.6 +/- 24.6% increase in the total charge flux carried by single inhibitory postsynaptic currents. This effect may be due to prolonged activation of postsynaptic GABA(A) receptors by excess GABA released in response to increased excitation. Exposure to the GABA uptake blocker, nipecotic acid, similarly prolonged current decay without affecting the maximal peak conductance. Our findings suggest that the limit on recruitment of evoked fast inhibition in neocortical layer V pyramidal cells arises from the saturation of postsynaptic GABA(A) receptors. However, while there is a limit to the peak fast inhibitory postsynaptic conductance which can be recruited with increasing excitation, inhibitory strength may still be modulated by increasing charge flux through the prolongation of fast inhibition.

Animals↗

PubMed1997

Recruitment of GABAergic inhibition and synchronization of inhibitory interneurons in rat neocortex.

Intracellular recordings were obtained from pyramidal and interneuronal cells in rat neocortical slices to examine the recruitment of GABAergic inhibition and inhibitory interneurons. In the presence of the convulsant agent 4-aminopyridine (4-AP), after excitatory amino acid (EAA) ionotropic transmission was blocked, large-amplitude triphasic inhibitory postsynaptic potentials (IPSPs) occurred rhythmically (every 10-40 s) and synchronously in pyramidal neurons. After exposure to the gamma-aminobutyric acid-A (GABA(A)) receptor antagonist picrotoxin, large-amplitude monophasic slow IPSPs persisted in these cells. In the presence of 4-AP and EAA blockers, interneurons showed periodic spike firing. Although some spikes rode on an underlying synaptic depolarization, much of the rhythmic firing consisted of spikes having highly variable amplitudes, arising abruptly from baseline, even during hyperpolarization. The spike firing and depolarizing synaptic potentials were completely suppressed by picrotoxin exposure, although monophasic slow IPSPs persisted in interneurons. This suggests that this subset of interneurons may participate in generating fast GABA(A) IPSPs, but not slow GABA(B) IPSPs. Cell morphology was confirmed by intracellular injection of neurobiotin or the fluorescent dye Lucifer yellow CH. Dye injection into interneurons often (>70%) resulted in the labeling of two to six cells (dye coupling). These findings suggest that GABA(A)ergic neurons may be synchronized via recurrent collaterals through the depolarizing action of synaptically activated GABA(A) receptors and a mechanism involving electrotonic coupling. Although inhibitory neurons mediating GABA(B) IPSPs may be entrained by the excitatory GABA(A) mechanism, they appear to be a separate subset of GABAergic neurons capable of functioning independently with autonomous pacing.

Animals↗

PubMed1997

Epileptogenesis following neocortical trauma from two sources of disinhibition.

Epileptogenesis following neocortical trauma from two sources of disinhibition. J. Neurophysiol. 78: 2804-2810, 1997. Intracellular and field potential recordings were obtained from superficial and deep neurons from both intact coronal rat somatosensory slices, and from slices which had been acutely divided into a superficial strip of cortex ( approximately 450 micron from the pia) and a deep segment. Membrane properties for cells in the traumatized slices were similar to those of their counterparts in intact slices. However, synaptic hyperexcitability developed in the deep segments in which a majority of cells likely underwent dendrotomy. This hyperexcitability was manifested by epileptiform activity in 54% of traumatized slices. Measurements of fastGABAergic inhibitory strength showed these slices were disinhibited. Superficial delivery of tetrodotoxin to the upper 450 micron of intact slices led to disinhibition of fast GABAergic transmission as well as an attendant increase in excitatory postsynaptic potential strength but not epileptogenesis. Pharmacological maneuvers aimed at preventing glutamate-triggered increases in intracellular calcium [glutamate ionotropic antagonists, dantrolene, and bis-(o-aminophenoxy)-N,N,N', N'-tetraacetic acid (BAPTA)-AM] showed that a 1 h treatment in these agents conferred protection against epileptogenesis. These results demonstrate that the seizure-like activity developing in deep dendrotomized cortical segments resulted from two sources of GABAergic disinhibition: the physical removal of important superficial inhibitory circuits and glutamate-triggered increases in intracellular calcium.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1995

Activity-dependent depression of monosynaptic fast IPSCs in hippocampus: contributions from reductions in chloride driving force and conductance.

Whole-cell recordings techniques were used to record pharmacologically isolated fast inhibitory postsynaptic currents (IPSCs) in CA1 pyramidal neurons from rat hippocampal slices. Repetitive extracellular stimulation up to 10 Hz progressively reduced steady-state fast IPSC amplitude. At low stimulation frequencies (up to 1 Hz), this attenuation was characterized by a positive shift of IPSC reversal potential with no change in IPSC conductance. Above 1 Hz stimulation, fast IPSC depression was associated with changes in both reversal potential and IPSC conductance. Use-dependent depression at low frequencies was prevented when cells were chloride-loaded using cesium chloride based intracellular solutions. These findings suggest that activity-dependent depression of fast IPSCs at low stimulus frequencies results entirely from a reduction in chloride driving force, stemming from intracellular chloride accumulation. Activity-dependent changes in fast IPSC conductance occur only at stimulation rates above 1 Hz.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1995

N-methyl-D-aspartate transmission modulates GABAB-mediated inhibition of rat hippocampal pyramidal neurons in vitro.

Slow inhibition was investigated by stimulating inhibitory neurons at the border of stratum radiatum and lacunosum-moleculare with focal microapplications of glutamate, while recording resultant slow inhibitory postsynaptic potentials in CA1 pyramidal neurons in rat hippocampal slices. The slow inhibitory postsynaptic potentials evoked had an average peak amplitude of -2.2 mV, measured at -60 mV. Their peak conductance was 2.5 nS. These events were characterized as slow GABAB inhibitory postsynaptic potentials because they reversed at -90 mV, and were blocked by CGP 35348 (500 microM). Exposure to magnesium-free solutions augmented glutamate-evoked slow inhibitory postsynaptic potentials. Mean peak amplitude and conductance were -3.1 mV and 4.0 nS. Exposure to the N-methyl-D-aspartate antagonist MK-801 (20 microM) allowed separation of the glutamate-triggered slow inhibitory postsynaptic potential into components induced by non-N-methyl-D-aspartate and N-methyl-D-aspartate receptor activation. The N-methyl-D-aspartate component dominated, even under control conditions, and could account for up to 60% of the control slow inhibitory postsynaptic potential. Thus, the activation and recruitment of GABAB-mediated inhibition depend on both non-N-methyl-D-aspartate and N-methyl-D-aspartate-mediated excitation of inhibitory interneurons. Under physiological conditions slow inhibition may act as an important synaptic filtering mechanism, but when N-methyl-D-aspartate-mediated excitation increases, slow inhibition is further recruited, providing an important means to offset excessive excitation.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1995

An in vitro model of persistent epileptiform activity in neocortex.

An in vitro model of persistent epileptiform activity was developed to study the mechanisms involved in epileptogenesis. Extracellular recordings were obtained from rat neocortical slices exposed to magnesium-free solution for 2 h. During exposure to magnesium-free solution spontaneous epileptiform activity consisting of interictal bursting and ictal-like discharges were observed. Interestingly, this activity persisted for hours after the slices were returned to magnesium-containing control solution. The N-methyl-D-aspartate (NMDA) receptor antagonist CPP prevented the development of the epileptiform activity, while the non-NMDA receptor antagonist CNQX abolished the epileptiform discharge that persisted after slices were returned to control solution. These findings suggest there are two distinct phases in the development of epileptic activity in this model, namely, induction (mediated by NMDA receptor activity) and maintenance (supported largely by non-NMDA receptor activity). The similarities and possible parallels between the mechanisms underlying this epileptogenesis and other forms of use-dependent modification of synaptic excitation, such as long-term potentiation, are discussed. This in vitro model of neocortical epileptogenesis may provide insights into the events underlying the development of clinical partial epilepsy.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1995

Recruitment of GABAA inhibition in rat neocortex is limited and not NMDA dependent.

1. The recruitment of evoked fast inhibitory postsynaptic currents (IPSCs) and excitatory postsynaptic currents (EPSCs) was examined using whole cell voltage-clamp recordings from layer V pyramidal neurons in slices of rat somatosensory cortex. Synaptic currents were evoked with graded electrical stimulation to assess the relative activation of IPSCs and EPSCs. Fast GABAA ergic IPSCs were selectively recorded by holding cells at potentials equal to EPSC reversal (approximately 0 mV). EPSCs were likewise isolated by holding cells at IPSC reversal potential (about -75 mV). 2. As stimulus intensities were increased, the magnitude of the postsynaptic currents also increased. Over the range of stimuli applied (2-10 V), EPSCs did not exhibit an upper limit. However, fast gamma-aminobutyric acid-A (GABAA-mediated IPSCs reached a maximum at intensities approximately 2 times threshold. 3. The limit on fast inhibition was unresponsive to alterations in N-methyl-D-aspartate (NMDA)-mediated excitation. Exposure to nominally magnesium-free solutions or to the NMDA antagonist 3-[(RS)-2-carboxypiperazin-4-yl]-propyl-1-phosphonic acid did not affect the fast IPSC maximum. Shifts in the input-output curves for submaximal activation of IPSCs were seen, which were attributed to polysynaptic excitation. 4. Blockade of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate (non-NMDA) receptors with 6-cyano-7-nitro-quinoxaline-2,3-dione (CNQX) completely abolished synaptically driven, fast GABAA-mediated inhibition. These findings suggested that neocortical inhibitory cells could be driven exclusively through non-NMDA transmission. 5. By comparison, in hippocampal CA1 pyramidal neurons maximal fast inhibition was sensitive to both NMDA and non-NMDA receptor blockade. 6. The results in neocortex were corroborated by direct intracellular recordings from layer V-VI interneurons. Non-NMDA receptor blockade with CNQX prevented synaptic activation of action potentials in these cells, even during cotreatment with magnesium-free solution. 7. Together, these results suggest that recruitment of GABA(A) ergic IPSCs in neocortex is ultimately driven via glutamatergic afferents arriving at non-NMDA receptors on interneurons. Properties limiting fast inhibition would favor the propagation of enhanced excitatory activity through the neuronal network.

Animals↗

PubMed1994

Separate activation of fast and slow inhibitory postsynaptic potentials in rat neocortex in vitro.

Synaptic inhibition was investigated by stimulating inhibitory neurones with focal microapplications of glutamate, while recording from layer V pyramidal neurones of rat somatosensory cortical slices. One class of inhibitory postsynaptic potentials (IPSPs) thus elicited was characterized as a fast, chloride-mediated, GABAA IPSP in part by its fast time-to-peak (mean 2.5 ms) and brief duration, but primarily on the basis of its reversal potential at -68 mV, and its blockade by picrotoxin. The average peak amplitude for these fast IPSPs was -1.5 mV, measured at -60 mV. The peak conductance calculated for these events was about 10 nS. The conductance change associated with the maximal fast inhibitory postsynaptic potential resulting from electrical stimulation of afferent pathways ranged up to 116 nS. A second class of IPSP was encountered much less frequently. These glutamate-triggered events were characterized as slow, potassium-mediated GABAB IPSPs partly because of their longer times-to-peak (mean, 45 ms) and duration, but especially because of their extrapolated equilibrium potential at about -89 mV and blockade by 2-hydroxysaclofen. The average peak amplitude for these slow IPSPs was -2.3 mV, measured at -60 mV. The peak conductance for these events was about 8 nS. IPSPs resulting from the excitation of individual inhibitory interneurones were elicited by glutamate microapplication at particular locations relative to recording sites. Both fast and slow IPSPs were generated, but these occurred as separate events, and mixed responses were never seen. Thus, the two mechanistically distinct types of IPSPs which result from GABA interaction at GABAA and GABAB receptors on neocortical neurones may be mediated by separate classes of inhibitory neurones.

Animals↗

PubMed1994

Properties of isolated GABAB-mediated inhibitory postsynaptic currents in hippocampal pyramidal cells.

Whole-cell recording techniques were used to record isolated slow inhibitory postsynaptic currents in CA1 pyramidal neurons from rat hippocampal slices. Application of 6-cyano-7-nitroquinoxaline-2,3-dione and 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid eliminated excitatory synaptic transmission, resulting in a 38% reduction in slow inhibitory postsynaptic current magnitude. Subsequent addition of the GABAA antagonist picrotoxin caused a further decrease in slow inhibitory postsynaptic current amplitude. The remaining, isolated slow inhibitory postsynaptic current was blocked by the GABAB antagonist 2-hydroxysaclofen and when cesium was substituted for intracellular potassium. The kinetics of isolated slow inhibitory postsynaptic currents were characterized by single exponential, fourth power activation, and double exponential inactivation. These slow inhibitory postsynaptic currents had a reversal potential of -85.7 +/- 1.6 mV, and a slope conductance of 935 +/- 277 pS. Single slow inhibitory postsynaptic currents carried a total charge flux of 13.4 +/- 7.6 pC. Repetitive stimulation up to 1 Hz progressively reduced steady-state slow inhibitory postsynaptic current amplitude. This attenuation was characterized by a decrease in slope conductance, but slow inhibitory postsynaptic current reversal potential remained unchanged, as did slow inhibitory postsynaptic current kinetics. These results indicate that, under physiological conditions, both ionotropic glutamate- and GABAA-mediated transmission contribute to slow inhibitory postsynaptic current recruitment. Given this finding, activity-dependent decreases in GABAA transmission could contribute to slow inhibitory postsynaptic current depression, though not exclusively, since isolated slow inhibitory postsynaptic currents also demonstrated this property. The use-dependent depression of isolated slow inhibitory postsynaptic currents may be a consequence of a reduction in transmitter release.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1993

Recruitment of inhibition by enhanced activation of synaptic NMDA responses in the rat cerebral cortex.

Intracellular recordings of layer V neurons from rat neocortical slices were obtained to examine the effects of reducing extracellular magnesium on inhibition. Magnesium-free solutions induced interictal and ictal-like events in cortical neurons. Changes in synaptic events underlying epileptogenesis were studied when extracellular calcium was raised (from 2 to 3-7 mM) since this delayed seizure activity. With increasing time of exposure of cells to magnesium-free solutions, there was a significant increase in the size and duration of both the depolarizing and slow synaptic hyperpolarizing responses, but the fast synaptic hyperpolarization significantly declined in amplitude. When cells were recorded with cesium acetate-filled microelectrodes slow hyperpolarizing responses were blocked, but depolarization of cells to 0 mV allowed an isolated fast hyperpolarizing response to be recorded following synaptic stimulation. The amplitude of this response was unchanged after exposure to magnesium-free solutions. Synaptic responses of cells initially bathed in an N-methyl-D-aspartate (NMDA) antagonist (CPP) were unchanged by subsequent exposure to magnesium-free solutions. CPP exposure by itself caused a decrease in depolarization duration, increase in fast hyperpolarizing amplitude, and decrease in slow hyperpolarization amplitude and duration. When the fast hyperpolarization was viewed in isolation (cesium recording electrodes) at 0 mV, the amplitude of this event was unchanged by exposure to CPP. Given these results stimulus-response characteristics of neocortical neurons were reassessed under control conditions. With higher intensity stimuli larger depolarizing and slow hyperpolarizing responses were evoked, but the fast hyperpolarization showed a decremental response. These effects were reversed when CPP was added. When NMDA activity was enhanced by exposure to magnesium-free solutions or electrical stimulation, the amplitude of excitatory events and slow hyperpolarizations increased, but fast inhibitory responses showed limited capacity for incremental recruitment. This suggests fast inhibition is saturated (maximal) at submaximal levels of excitation, and can be overcome by increasing levels of excitation. Such a process is active under physiological conditions, altering the efficacy of inhibition.

Animals↗

PubMed1993

GABAA receptor-mediated mechanisms contribute to frequency-dependent depression of IPSPs in the hippocampus.

Intracellular recordings from in vitro guinea-pig hippocampal slices were obtained to investigate the mechanisms underlying activity-dependent depression of inhibitory postsynaptic potentials (IPSPs) in CA1 pyramidal neurons. IPSPs were studied following blockade of glutamatergic transmission. Attenuation of both fast and slow IPSP components was apparent at stimulus rates of > 0.2 Hz, but the late IPSP showed depression at even lower rates of stimulation. Overlap of these events made resolving response components during depression difficult. Fast IPSPs were isolated using CsCl-filled microelectrodes, blocking slow potassium-dependent IPSPs and reversing the chloride gradient. Under this condition repetitive stimulation did not induce IPSP depression. Other experiments showed slow IPSP depression at stimulus rates of < 0.5 Hz was relieved in the presence of picrotoxin which antagonizes GABAA events. These results suggest that activity-dependent depression of the fast IPSP results largely from chloride accumulation as a consequence of repetitive activation. Depression of the slow IPSP appears to arise from at least two components: (1) a masking of the slow hyperpolarization resulting from an apparent increase in the depolarizing GABA response due to chloride accumulation; and (2) a novel process involving GABAA receptors, perhaps mediated through presynaptic inhibition.

Amino Acids↗

PubMed1993

Characterization of cholinergic and noradrenergic slow excitatory postsynaptic potentials from rat cerebral cortical neurons.

Intracellular recordings from layer V pyramidal neurons in rat somatosensory neocortical slices were used to investigate the effects of electrically stimulating slices known to contain cholinergic and noradrenergic fibers. Repetitive electrical stimulation ventral to the recording site elicited a series of fast excitatory postsynaptic potentials followed by an inhibitory postsynaptic potential. These potentials were followed by a slow excitatory postsynaptic potential that lasted up to tens of seconds. The slow excitatory postsynaptic potential was more prominent when neurons were depolarized to 5-10 mV below firing threshold and was associated with increased input resistance and generated action potentials. The slow excitatory postsynaptic potential increased the amplitude of membrane potential oscillations and blocked the slow afterhyperpolarization which followed trains of action potentials. The amplitude of the slow excitatory postsynaptic potential was sensitive to extracellular potassium concentration. Blockade of postsynaptic action potentials by QX-314 did not block slow excitatory postsynaptic potentials. Exposure of slices to tetrodotoxin did block slow excitatory postsynaptic potentials, indicating they were dependent on propagated action potentials. Application of antagonists of glutamate and fast GABA responses failed to block slow excitatory postsynaptic potentials. Exposure to atropine or either propranolol or atenolol partially antagonized slow excitatory postsynaptic potentials, but only when atropine was added in combination with one of the other agents was the slow excitatory postsynaptic potential completely blocked. Exposure of slices to eserine, imipramine, or cocaine enhanced slow excitatory postsynaptic potentials. It is concluded that the slow excitatory postsynaptic potential triggered in neocortical slices is a composite of a cholinergic and a noradrenergic slow excitatory postsynaptic potential, and these potentials are capable of altering the firing properties of neurons for tens of seconds.

Acetylcholine↗

PubMed1991

Acetylcholine and norepinephrine mediate slow synaptic potentials in normal and epileptic neocortex.

Slow excitatory postsynaptic potentials (EPSPs) were identified in rat neocortical slices. Such potentials, resistant to blockade of glutamate and gamma-aminobutyric acid-A (GABAA) receptors, were partially antagonized by muscarinic or beta-adrenergic antagonists separately, and completely blocked when these agents were added in combination. Slow EPSPs were enhanced by a cholinesterase inhibitor or catecholamine reuptake blockers. Spontaneous epileptic discharges induced by picrotoxin also triggered slow EPSPs. Such potentials were pharmacologically identical to those induced by electrical stimulation under normal conditions. A non-conventional mechanism for synaptic transmission is postulated to account for triggering of slow EPSPs by epileptic discharges.

Acetylcholine↗

PubMed1986

Oscillatory behavior in inferior olive neurons: mechanism, modulation, cell aggregates.

Inferior olive neurons, in brain slices maintained in vitro, display spontaneous, continuous oscillations of their membrane potential which are consonant with olivary rhythmic activity seen in vivo. This oscillatory behavior was studied with intracellular electrophysiological techniques. The 3-10 Hz rhythmicity of these cells from guinea pigs is tetrodotoxin resistant and dependent on a somatic calcium conductance. The oscillatory behavior can exhibit intrinsic frequency modulation and can be altered by synaptic processes. Synaptic alteration of the oscillatory behavior by afferent sources and extensive electrotonic coupling between cells in local aggregates (shown by Lucifer yellow dye-coupling) provide the substrate for a potent central pattern generator with a well established efferent pathway for control of motor functions.

Animals↗