Kinetics of GABAB autoreceptor-mediated suppression of GABA release in rat insular cortex

Release of GABA is controlled by presynaptic GABA receptor type B (GABA(B)) autoreceptors at GABAergic terminals. However, there is no direct evidence that GABA(B) autoreceptors are activated by GABA release from their own terminals, and precise profiles of GABA(B) autoreceptor-mediated suppression of GABA release remain unknown. To explore these issues, we performed multiple whole-cell, patch-clamp recordings from layer V rat insular cortex. Both unitary inhibitory and excitatory postsynaptic currents (uIPSCs and uEPSCs, respectively) were recorded by applying a five-train depolarizing pulse injection at 20 Hz. In connections from both fast-spiking (FS) and non-FS interneurons to pyramidal cells, the GABA(B) receptor antagonist CGP 52432 had little effect on the initial uIPSC amplitude. However, uIPSCs, responding to later pulses, were effectively facilitated. This CGP 52432-induced facilitation was prominent in the fourth uIPSCs, which were evoked 150 ms after the first uIPSC. The facilitation of uIPSCs was accompanied by an increase in the paired-pulse ratio. In addition, analysis of the coefficient of variation suggests the involvement of presynaptic mechanisms in CGP 52432-induced uIPSC facilitation. Paired-pulse stimulation (interstimulus interval = 150 ms) of presynaptic FS cells revealed that the second uIPSC was also facilitated by CGP 52432, which had little effect on the amplitude and interevent interval of miniature IPSCs. In contrast, uEPSCs, responding to all five stimulations of a presynaptic pyramidal cell, were less affected by CGP 52432. These results suggest that a single presynaptic action potential is sufficient to activate GABA(B) autoreceptors and to suppress GABA release in the cerebral cortex.     

D(2)-like dopamine receptors differentially regulate unitary IPSCs depending on presynaptic GABAergic neuron subtypes in rat nucleus accumbens shell

In the nucleus accumbens (NAc), a medium spiny (MS) neuron receives GABAergic inputs from two major sources: fast-spiking (FS) neurons and other, adjacent MS neurons. These two types of inhibitory synapses are considered to play different roles in output activities, i.e., FS→MS connections suppress output from the NAc whereas MS→MS connections contribute to lateral inhibition. In the present study, we focused on the electrophysiological properties of unitary inhibitory postsynaptic currents (uIPSCs) obtained from MS→MS connections and FS→MS connections and examined the effects of quinpirole, a dopamine D(2)-like receptor agonist, on uIPSCs with multiple whole cell patch-clamp recording. Application of quinpirole (1 μM) reliably suppressed the amplitude of uIPSCs by 29.6% in MS→MS connections, with increases in paired-pulse ratio and failure rate. The suppressive effects of quinpirole on uIPSCs were mimicked by 1 μM PD128907, a D(2/3) receptor agonist, whereas quinpirole-induced suppression of uISPCs was blocked by preapplication of 1 μM sulpiride or 10 μM nafadotride, both D(2/3) receptor antagonists. On the other hand, quinpirole (1 μM) had divergent effects on FS→MS connections, i.e., quinpirole increased uIPSC amplitude in 38.1% of FS→MS connections and 23.8% of FS→MS connections were suppressed by quinpirole. Analysis of coefficient of variation in uIPSC amplitude implied the involvement of presynaptic mechanisms in quinpirole-induced effects on uIPSCs. These results suggest that activation of D(2)-like receptors facilitates outputs from MS neurons in the NAc by reducing lateral inhibition during a dormant period of FS neuron activities.     

Burst firing induces a rebound of synaptic strength at unitary neocortical synapses

High-frequency activity produces transient depression at many synapses but also, as recently demonstrated, may accelerate the recovery from use-dependent depression. We have examined the possible consequences of this synaptic mechanism in neocortical excitatory synapses by recording simultaneously from presynaptic pyramidal neurons and their postsynaptic targets. Brief bursts of high-frequency spikes produced a strong depression of the amplitude of unitary excitatory postsynaptic currents (uEPSCs). However, when burst firing was combined with low-frequency ongoing activity, we found that the strong synaptic depression was followed by a transient rebound of synaptic strength. This rebound overshot the low-frequency baseline values and lasted 1-2 s. These results suggest that in the presence of ongoing activity, neocortical synapses may functionally facilitate following burst firing.     

Role for the subthreshold currents ILeak and IH in the homeostatic control of excitability in neocortical somatostatin-positive inhibitory neurons

Cortical circuitry reconfigures in response to chronic (1-3 days) changes in activity levels. To understand this process, we must know the role played by inhibitory neurons because they crucially influence network properties by controlling action potential generation and synaptic integration. Using pharmacological blockade of activity in neocortical organotypic slice cultures, we examined the activity-dependent regulation of membrane excitability in a specific inhibitory neuron subtype: the somatostatin-positive (SOM+) neuron. Chronic action potential blockade (TTX, 2.5 days) resulted in increased excitability in SOM+ neurons. This result is consistent with a homeostatic process to maintain the average firing rate of SOM+ neurons at a particular level. Excitability changes were not ascribed to changing cell size or alterations in voltage-dependent sodium current. Instead, the excitability increase was largely the result of a decrease in the density of two subthreshold currents: a passive leak current (ILeak) and H-current (IH). The downregulation of these currents increased excitability mostly through a decrease in membrane input conductance. The coadaptation of ILeak and IH enabled a change in input conductance while helping to preserve membrane potential. Evidence indicated that ILeak was probably mainly mediated by K+. At earlier culture ages, this adaptation was superimposed on developmental changes, whereas at older ages, the same types of induced alterations occurred but with no developmental component. Together with other studies, these data indicate that both inhibitory and excitatory neurons increase membrane excitability with chronic reduction in activity, but through different mechanisms.     

Synaptic inhibition of pyramidal cells evoked by different interneuronal subtypes in layer v of rat visual cortex

Properties of GABA(A) receptor-mediated unitary inhibitory postsynaptic currents (uIPSCs) in pyramidal (P) cells, evoked by fast spiking (FS) and low-threshold spike (LTS) subtypes of interneurons in layer V of rat visual cortex slices were examined using dual whole cell recordings. uIPSCs evoked by FS cells were larger and faster rising than those evoked by LTS cells, consistent with the known primary projections of FS and LTS cell axons to perisomatic and distal dendritic areas of layer V pyramidal cells, respectively, and the resulting electrotonic attenuation for LTS-P synaptic events. Unexpectedly, the decay time constants for LTS-P and FS-P uIPSCs were not significantly different. Modeling results were consistent with differences in the underlying GABA(A) receptor-mediated conductance at LTS-P and FS-P synapses. Paired-pulse depression (PPD), present at both synapses, was associated with an increase in failure rate and a decrease in coefficient of variation, indicating that presynaptic mechanisms were involved. Furthermore, the second and first uIPSC amplitudes during PPD were not inversely correlated, suggesting that PPD at both synapses is independent of previous release and might not result from depletion of the releasable pool of synaptic vesicles. Short, 20-Hz trains of action potentials in presynaptic interneurons evoked trains of uIPSCs with exponentially decreasing amplitudes at both FS-P and LTS-P synapses. FS-P uIPSC amplitudes declined more slowly than those of LTS-P uIPSCs. Thus FS and LTS cells, with their differences in firing properties, synaptic connectivity with layer V P cells, and short-term synaptic dynamics, might play distinct roles in regulating the input-output relationship of the P cells.     

Target-cell-specific facilitation and depression in neocortical circuits

In neocortical circuits, repetitively active neurons evoke unitary postsynaptic potentials (PSPs) whose peak amplitudes either increase (facilitate) or decrease (depress) progressively. To examine the basis for these different synaptic responses, we made simultaneous recordings from three classes of neurons in cortical layer 2/3. We induced repetitive action potentials in pyramidal cells and recorded the evoked unitary excitatory (E)PSPs in two classes of GABAergic neurons. We observed facilitation of EPSPs in bitufted GABAergic interneurons, many of which expressed somatostatin immunoreactivity. EPSPs recorded from multipolar interneurons, however, showed depression. Some of these neurons were immunopositive for parvalbumin. Unitary inhibitory (I)PSPs evoked by repetitive stimulation of a bitufted neuron also showed a less pronounced but significant difference between the two target neurons. Facilitation and depression involve presynaptic mechanisms, and because a single neuron can express both behaviors simultaneously, we infer that local differences in the molecular structure of presynaptic nerve terminals are induced by retrograde signals from different classes of target neurons. Because bitufted and multipolar neurons both formed reciprocal inhibitory connections with pyramidal cells, the results imply that the balance of activation between two recurrent inhibitory pathways in the neocortex depends on the frequency of action potentials in pyramidal cells.     

新生儿皮质第一层PV免疫反应Cajal-Retzius和非Cajal-Retzius神经元的分布

目的　研究Cajal Retzius(CR)和非Cajal Retzius(NCR)神经元在新生儿大脑 13个皮质区的分布、相应密度和可能的亚型。方法　采用免疫组织化学方法检测新生儿大脑皮质神经元数目及分布。结果　 (1)PV免疫反应阳性CR细胞见于所有的新皮质区 ,这些区域在第一层深部也发现PV ir水平纤维丛。 (2 )许多PV irCR细胞显示退行性变的明显标志。 (3)除大的CR细胞外 ,小的PV irNCR神经元也出现在新皮质区。它们包括不同的形态学种类 ,可以区分为几种亚型。结论　NCR细胞高密度地出现于第一感觉区 3、1、17和 4 1区。由于PV irNCR细胞密度在各区的差异 ,PV irCR与PV irNCR的比率在第一感觉区相对较低。目前对出生早期大鼠躯体感觉皮质的研究报道第一层存在复杂的相关的神经元间自发作用的时空模式。一个很大的可能性就是区域多样的神经元活性可能在不同脑区皮质环路中起主要作用。     

Microcircuits of excitatory and inhibitory neurons in layer 2/3 of mouse barrel cortex

Synaptic interactions between nearby excitatory and inhibitory neurons in the neocortex are thought to play fundamental roles in sensory processing. Here, we have combined optogenetic stimulation, whole cell recordings, and computational modeling to define key functional microcircuits within layer 2/3 of mouse primary somatosensory barrel cortex. In vitro optogenetic stimulation of excitatory layer 2/3 neurons expressing channelrhodopsin-2 evoked a rapid sequence of excitation followed by inhibition. Fast-spiking (FS) GABAergic neurons received large-amplitude, fast-rising depolarizing postsynaptic potentials, often driving action potentials. In contrast, the same optogenetic stimulus evoked small-amplitude, subthreshold postsynaptic potentials in excitatory and non-fast-spiking (NFS) GABAergic neurons. To understand the synaptic mechanisms underlying this network activity, we investigated unitary synaptic connectivity through multiple simultaneous whole cell recordings. FS GABAergic neurons received unitary excitatory postsynaptic potentials with higher probability, larger amplitudes, and faster kinetics compared with NFS GABAergic neurons and other excitatory neurons. Both FS and NFS GABAergic neurons evoked robust inhibition on postsynaptic layer 2/3 neurons. A simple computational model based on the experimentally determined electrophysiological properties of the different classes of layer 2/3 neurons and their unitary synaptic connectivity accounted for key aspects of the network activity evoked by optogenetic stimulation, including the strong recruitment of FS GABAergic neurons acting to suppress firing of excitatory neurons. We conclude that FS GABAergic neurons play an important role in neocortical microcircuit function through their strong local synaptic connectivity, which might contribute to driving sparse coding in excitatory layer 2/3 neurons of mouse barrel cortex in vivo.     

Contrasting of synaptic signals by simultaneous modification of excitatory and inhibitory inputs

Conditions facilitating long-term contrasting of interneuronal connections were studied using a mathematical model of posttetanic Ca²⁺-dependent postsynaptic processes in pyramidal neurons of hippocampal field CA₃. These studies demonstrated that modified inhibition selectively facilitates long-term potentiation of the efficiency of one of the interneuronal connections when the presynaptic neuron discharges at a given frequency for a short time, while connections formed from the same postsynaptic cell with other presynaptic neurons undergo long-term depression. The mechanism underlying this contrasting may involve long-term depression of the efficiency of disynaptic inhibitory transmission to the rhythmically stimulated input, even when the efficiency of monosynaptic excitatory transmission at the same input is low and undergoes minimal potentiation. When the “common” inhibitory neuron is simultaneously activated by various presynaptic cells, heterosynaptic potentiation of inhibitory transmission can simultaneously develop at the other inputs of the postsynaptic cell, without change in the efficiency of excitatory transmission, which leads to long-term depression of the efficiency of the connections between other excitatory neurons and the postsynaptic cell. Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, Moscow. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti, Vol. 46, No. 6, pp. 1076–1087, November–December, 1996.     

Presynaptic inactivation of action potentials and postsynaptic inhibition of GABAA currents contribute to KA-induced disinhibition in CA1 pyramidal neurons

Kainate-type glutamate ionotropic receptors (KAR) mediate either depression or potentiation of inhibitory transmission. The mechanisms underlying the depressant effect of KAR agonists have been controversial. Under dual patch-clamp recording techniques in synaptically coupled pairs of CA1 interneurons and pyramidal neurons in hippocampal slices, micromolar concentrations of KAR agonists, kainic acid (KA, 10 microM) and ATPA (10 microM), induced inactivation of action potentials (APs) in 58 and 50% of presynaptic interneurons, respectively. Inactivation of interneuronal APs might have significantly contributed to KA-induced decreases in evoked inhibitory postsynaptic currents (eIPSCs) that are obtained by stimulating the stratum radiatum. With controlled interneuronal APs, KAR agonists induced a decrease in the potency (mean amplitude of successful events) and mean amplitude (including failures) of unitary inhibitory postsynaptic currents (uIPSCs) without significantly changing the success rate (P(s)) at perisomatic high-P(s) synapses. In contrast, KAR agonists induced a decrease in both the P(s) and potency of uIPSCs at dendritic high-P(s) synapses. KAR agonists induced an inhibition of GABA(A) currents by activating postsynaptic KARs in pyramidal neurons; this was more prominent at dendrites than at soma. Both the exogenous GABA-induced current and the amplitude of miniature IPSCs (mIPSCs) were attenuated by KAR agonists. Thus the postsynaptic KAR-mediated inhibition of GABA(A) currents may contribute to the KAR agonist-induced decrease in the potency of uIPSCs and KA-induced disinhibition.     

Network activity evoked by neocortical stimulation in area 36 of the guinea pig perirhinal cortex.

The perirhinal cortex is a key structure involved in memory consolidation and retrieval. In spite of the extensive anatomical studies that describe the intrinsic and extrinsic associative connections of the perirhinal cortex, the activity generated within such a network has been poorly investigated. We describe here the pattern of synaptic interactions that subtend the responses evoked in area 36 of the perirhinal cortex by neocortical and local stimulation. The experiments were carried out in the in vitro isolated guinea pig brain. The synaptic perirhinal circuit was reconstructed by integrating results obtained during intracellular recordings from layer II-III neurons with simultaneous current source density analysis of laminar profiles performed with 16-channel silicon probes. Both neocortical and local stimulation of area 36 determined a brief monosynaptic excitatory potential in layer II-III neurons, followed by a biphasic synaptic inhibitory potential possibly mediated by a feed-forward inhibitory circuit at sites close to the stimulation electrode and a late excitatory postsynaptic potential (EPSP) that propagated at distance within area 36 along the rhinal sulcus. During a paired-pulse stimulation test, the inhibitory postsynaptic potential (IPSP) and the late EPSP were abolished in the second conditioned response, suggesting that they are generated by poli-synaptic circuits. Current source density analysis of the field responses demonstrated that 1) the monosynaptic activity was generated in layers II-III and 2) the sink associated to the disynaptic responses was localized within the superficial layer of area 36. We conclude that the neocortical input induces a brief monosynaptic excitation in area 36 of the perirhinal cortex, that is curtailed by a prominent inhibition and generates a recurrent excitatory associative response that travels at distance within area 36 itself. The results suggest that the perirhinal cortex network has the potentials to integrate multimodal incoming neocortical information on its way to the hippocampus.     

Carbenoxolone modifies spontaneous inhibitory and excitatory synaptic transmission in rat somatosensory cortex

Gap junction (GJ) coupling between neocortical GABAergic interneurons plays a critical role in the synchronization of activity in cortical networks in physiological and pathophysiological states, e.g., seizures. Past studies have shown that GJ blockers exert anticonvulsant actions in both in vivo and in vitro models of epilepsy. However, the precise mechanisms underlying these antiepileptic effects have not been fully elucidated. This is due, in part, to a lack of information of the influence of GJ blockade on network activity in the absence of convulsant agents or enhanced neuronal excitation. One key question is whether GJ blockers act on excitatory or inhibitory systems, or both. To address this issue, we examined the effects of the GJ blocker carbenoxolone (CarbX, 150 microM) on spontaneous inhibitory postsynaptic currents (sIPSCs) and excitatory postsynaptic currents (sEPSCs) in acute slices of rat somatosensory cortex. Results showed that CarbX decreased the amplitude and frequency of sIPSCs by 30.2% and 25.7%, respectively. CarbX increased the mean frequency of sEPSCs by 24.1%, but had no effect on sEPSC amplitude. During blockade of GABAA-mediated events with picrotoxin (20 microM), CarbX induced only a small increase in sEPSC frequency that was not statistically different from control, indicating CarbX enhancement of sEPECs was secondary to the depression of synaptic inhibition. These findings suggest that in neocortex, blockade of GJs leads to an increase in spontaneous excitation by uncoupling GABAergic interneurons, and that electronic communication between inhibitory cells plays a significant role in regulating tonic synaptic excitation.     

Distinct local circuits between neocortical pyramidal cells and fast-spiking interneurons in young adult rats

Connections between layer V pyramidal cells and GABAergic fast-spiking interneurons (pyramidal-FS) were studied by paired recordings combined with morphological analyses in acute neocortical slices from 28- to 52-day-old rats. Pairs of spikes elicited in pyramidal cells at a stimulation rate of 0.2 Hz induced unitary excitatory postsynaptic currents (EPSCs) in FS interneurons that displayed facilitation (48%), depression (38.5%), or neither depression nor facilitation (13.5%). Analyses of the EPSC amplitude distributions indicate that depressing connections always showed multiple functional release sites. On the contrary, facilitating connections consisted either of one or several release sites. At a holding potential of -72 mV, the quantal size (q) and the release probability (p) of facilitating connections with a single release site were -21.9 +/- 7.5 pA and 0.49 +/- 0.19 (SD), respectively. The mean q and the estimated number of release sites (n) at connections showing multiple sites were obtained by decreasing the release probability and did not differ between depressing and facilitating synapses (depressing connections: q = -15.3 +/- 2.5 pA, n = 5.1 +/- 3, facilitating connections: q = -23.9 +/- 9.8 pA, n = 7.8 +/- 5.4). However, the quantal content at facilitating synapses with multiple sites (1.9 +/- 1.5) was significantly different from that at depressing connections (4.1 +/- 3.9). Finally, quantitative morphological analyses revealed that most of the pyramidal cells displaying facilitation can be differentiated from those displaying depression by a more densely branched apical dendritic tree. Therefore two types of morphologically distinct pyramidal cells form excitatory connections with FS interneurons that differ in their short-term plasticity characteristics. Facilitating and depressing connections may provide a differential control of the temporal information processing of FS cells and thus finely regulate the inhibitory effect of these interneurons in neocortical networks of young adult rats.     

Activity- and BDNF-induced plasticity of miniature synaptic currents in ES cell-derived neurons integrated in a neocortical network

In vitro differentiated embryonic stem (ES) cells have been proposed as potential donor cells for cell replacement therapies of neurodegenerative diseases. The functional synaptic integration of such cells appears conceivable because ES cell-derived neurons are well known to establish excitatory and inhibitory synapses. However, long-term synaptic plasticity, a prerequisite of memory formation, has not yet been demonstrated at these synapses. After in vitro differentiation and purification by immunoisolation, we co-cultured ES cell-derived neurons with neocortical explants, which strongly innervated the ES cell-derived target neurons. ES cell-derived neurons exhibited action potential firing similar to primary cultured neocortical neurons. The formation of glutamatergic synapses was indicated by AMPA receptor-mediated miniature excitatory postsynaptic currents (AMPA mEPSCs). In addition, a N-methyl-D-aspartate receptor-mediated, D-2-amino-5-phosphonopentanoic acid-sensitive mEPSC component was observed. We first studied activity-dependent homeostatic plasticity (synaptic scaling) of mEPSCs at glutamatergic synapses. Chronic blockade of action potential activity by TTX resulted in an increase in the amplitudes of AMPA mEPSCs. This indicates that ES cell-derived neurons are capable of a homeostatic regulation of postsynaptic AMPA receptors. In addition, we investigated neurotrophin-induced synaptic plasticity of mEPSCs at glutamatergic synapses. Chronic addition of brain-derived neurotrophic factor (BDNF; 100 ng/ml) to the culture medium resulted in an increase in both the frequency and the amplitudes of AMPA mEPSCs. These results suggest that BDNF induces the formation and/or the functional maturation of presynaptic release sites in parallel with an upregulation of postsynaptic AMPA receptors. Thus BDNF represents a potential co-factor that could improve functional synaptic integration of ES cell-derived neurons into neocortical networks.     

Simultaneous recording of presynaptic spikes and excitatory postsynaptic potentials from monosynaptically connected hippocampal neurons

A technique has been devised to activate single granule cells in the hippocampus, and to record simultaneously spikes from the particular granule cell and excitatory postsynaptic potentials from a monosynaptically connected CA3 neuron. The unitary excitatory postsynaptic potentials (EPSPs) sustained for long observation periods, and increased in size with increases in stimulus frequency and in external Ca2+ concentration. This technique may be useful for quantal analysis of transmission through the synapse between mossy fibers and CA3 neurons.     

Long-term changes in the efficiency of inhibitory transmission in the thalamocortical neuronal networks induced by microstimulation of the cortex

Long-term potentiation (LTP) and long-term depression (LTD) of the efficiency of inhibitory transmission between different elements of a network took place in neuronal networks consisting of AC and MGB cells, as a result of MS of the cortex. LTP of inhibition was an input-specific effect, since it could develop simultaneously with LTP of the efficiency of excitatory transmission to the same cell and do not lead to a decrease in its baseline frequency. LTP (LTD) of inhibition was observed simultaneously with an increase (decrease) in the frequency of baseline impulse activity of the inhibitory neuron or neuron which is presynaptic in relation to an inhibitory neuron; the efficiency of the synaptic effect of one inhibitory neuron on various postsynaptic cells could vary in different directions. The data obtained may suggest the participation of both pre- and postsynaptic mechanisms in the modification of the efficiency of inhibition. Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, Moscow. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 45, No. 3, pp. 538–550, May–June, 1995.     

Analysis of the Excitatory and Inhibitory Components of Postsynaptic Currents Recorded in Pyramidal Neurons and Interneurons in the Rat Hippocampus

Postsynaptic currents recorded from interneurons and pyramidal cells in hippocampal slices by local voltage clamping were found to be the sum of excitatory (EPSC) and inhibitory (IPSC) components. An approach allowing quantitative assessment of the amplitude and time course of EPSC and IPSC without pharmacological blockade of the major postsynaptic receptors involved in generating these currents was developed. The approach is based on the existence of a significant difference between reversion potentials of cationic and anionic currents and the presence of a linear zone in the voltage-current characteristics of responses to excitatory and inhibitory transmitters. Comparison of the results of this calculation-based method with those of classical pharmacological analysis of the excitatory and inhibitory components of postsynaptic currents showed them to be virtually identical, which allows synaptic currents in defined neurons to be studied without altering the state of synaptic connections throughout the brain slice. IPSC was found to make a smaller contribution to the total postsynaptic current recorded in interneurons as compared with pyramidal neurons in rat hippocampal field CA1.__________Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 90, No. 8, pp. 945–956, August, 2004.     

Interactions and Firing Patterns of Rabbit Amygdalar Neurons in Unconditioned Fear Manifested as Freezing

The characteristics of the operation of the amygdalar neural network in unconditioned fear were identified by studying the interactions and nature of the spike activity of individual neurons in the basal and central nuclei of the amygdala in rabbits during freezing (fear), active unconditioned motor reactions (absence of fear), and in response to emotionally significant stimuli, as well as in calm waking. When rabbits froze, there were specific changes in the interactions of close-lying amygdalar neurons as compared with other states; these changes were not seen in the spike activity of individual neurons. Freezing increased the numbers of short-latency (up to 100 msec) excitatory connections and decreased the numbers of long-latency (250–450 msec) inhibitory connections. Neuron interactions were seen at frequencies in the delta2 range (2–4 Hz) more often in this state than in others. When the animals made active motor responses to the stimulus, there were decreases in the numbers of interacting neurons and increases in the numbers of longlatency (200–250 msec) excitatory and short-latency (50–200 msec) inhibitory connections, and a greater proportion of interactions occurred at frequencies of the theta1 range more frequently than in other states. Thus, the balance between the excitatory and inhibitory components of the amygdalar neural network is important for the occurrence of fear.     

Properties of depolarization-induced suppression of inhibitory transmission in cultured rat hippocampal neurons

 Properties of depolarization-induced suppression of inhibitory transmission (”DSI”) in cultured rat hippocampal neurons were examined, by recording inhibitory postsynaptic currents (IPSCs) evoked by single presynaptic neurons. In about 40% of the inhibitory synapses, transient suppression of IPSCs was induced by applying a depolarizing pulse (to 0 mV, > 2 s) to the postsynaptic neuron, which was identified to be γ-aminobutyric acid dependent (GABAergic) in some pairs, and to be glutamatergic in some other pairs. This depolarization-induced suppression of IPSCs, ”DSI”, was Ca²⁺ dependent, and was associated with an increase in the paired-pulse ratio. Phorbol esters had an occluding effect on DSI when applied to the bath solution, but not to the pipette solution used for the postsynaptic neuron. Application of the opioid receptor antagonist naloxone had no effect on DSI. The present study demonstrates that a pair of cultured hippocampal neurons can exhibit DSI similar to that previously reported to occur in hippocampal slices; this suggests that a dissociated cell culture system can provide a useful model system for the study of DSI. Furthermore, it was found that inhibitory neurons, in addition to excitatory ones, can induce DSI, and that a process sensitive to phorbol esters, but not activation of opioid receptors, is involved in DSI. Received: 20 May 1997 / Accepted: 1 September 1997     

Role of protein kinase C in modulation of excitability of CA1 pyramidal neurons in the rat

Biochemical and in situ hybridization studies demonstrated that the levels of protein kinase C variants were significantly increased in the hippocampus of the experimental models of epilepsy in rats. In addition it has been demonstrated that protein kinase C plays an important role in modulating synaptic transmission in the hippocampus. We examined the effects of activating of protein kinase C on the excitability of CA1 pyramidal neurons and synaptic transmission, using whole-cell current-clamp and extracellular field potential recording techniques. Indolactam V (1 microM) a novel protein kinase C activator, increased the excitability of CA1 neurons acting at both pre- and post-synaptic sites. Indolactam V, acting postsynaptically, significantly reduced the threshold for initiation of action potential from -42+/-3.8 mV to -51+/-3.1 mV and selectively inhibited the slow afterhyperpolarizing potential. Indolactam V also altered the neuronal firing properties in response to prolonged depolarizing pulse by eliminating the spike frequency accommodation. Our data indicate that indolactam V potentiated both amplitudes of Shaffer-collateral stimulation evoked excitatory postsynaptic currents and disynaptically evoked inhibitory evoked postsynaptic currents. However, the potentiation of inhibitory evoked postsynaptic currents amplitudes was not observed after blockade of NMDA and AMPA/kainate currents suggesting it was due to excitatory activity driving inhibitory neurons. The results indicate that the potentiation of pharmacologically isolated excitatory postsynaptic currents (215% of control) and amplitudes of population spikes (290% of control) was due to action of indolactam V presynaptically since the agonist reduced the paired-pulse ratio and the potentiating effect was not blocked by dialyzing the postsynaptic neuron through the recording electrode with a specific protein kinase C inactivator calphostin C. These findings suggest that protein kinase C increases the amplitude of epileptiform activity by causing potentiation of excitatory synaptic transmission, increasing the excitability of postsynaptic neurons and reducing negative feed back provided by slow afterhyperpolarizing potential.