Oxidative stress induced by cumene hydroperoxide produces synaptic depression and transient hyperexcitability in rat primary motor cortex neurons.

Pyramidal neurons of the motor cortex are selectively degenerated in Amyotrophic Lateral Sclerosis (ALS). The mechanisms underlying neuronal death in ALS are not well established. In the absence of useful biomarkers, the early increased neuronal excitability seems to be the unique characteristic of ALS. Lipid peroxidation caused by oxidative stress has been postulated as one of the possible mechanisms involved in degeneration motor cortex pyramidal neurons. This paper examines the effect of lipid peroxidation on layer V pyramidal neurons induced by cumene hydroperoxide (CH) in brain slices from wild type rats. CH induces a synaptic depression of pyramidal neurons in a time dependent manner, already observable on GABAergic synaptic transmission after 5min application of the drug. Altogether, our whole-cell patch-clamp recording data suggest that the functional changes induced by CH upon pyramidal neurons are due to pre- and postsynaptic mechanisms. CH did not alter mEPSCs or mIPSCs, but decreased the frequency, amplitude, and decay rate of spontaneous EPSCs and IPSCs. These effects may be explained by a presynaptic mechanism causing a decrease in action potential-dependent neurotransmitter release. Additionally, CH induced a postsynaptic inward current that underlies a membrane depolarization. Depressing the input flow from the inhibitory premotor interneurons causes a transient hyperexcitability (higher resistance and lower rheobase) in pyramidal neurons of the motor cortex by presumably altering a tonic inhibitory current. These findings, which resemble relevant cortical pathophysiology of ALS, point to oxidative stress, presumably by lipid peroxidation, as an important contributor to the causes underlying this disease.

Protein phosphatase-mediated regulation of protein kinase C during long-term depression in the adult hippocampus in vivo.

The neural substrates of learning and memory are thought to involve use-dependent long-term changes in synaptic function, including long-term depression (LTD) of synaptic strength. One biochemical event hypothesized to contribute to the maintenance and expression of LTD is decreased protein phosphorylation, caused by a decrease in protein kinase activity and/or an increase in protein phosphatase activity. We tested whether the activity of protein kinase C (PKC) decreases after the induction of LTD in area CA1 of the adult hippocampus in vivo, and then investigated the mechanism responsible for the LTD-associated alteration in PKC activity. We found that LTD was associated with a significant decrease in both autonomous and cofactor-dependent PKC activity. The decrease in PKC activity was prevented by NMDA receptor blockade and was not accompanied by a decrease in the level of either PKCalpha, beta, gamma, or zeta. Western blot analysis with phosphospecific antibodies revealed that phosphorylation of Ser-657 on the catalytic domain of PKCalpha (Ser-660 on PKCbetaII) was decreased significantly after the induction of LTD, and that this dephosphorylation was prevented by the protein phosphatase inhibitor okadaic acid. The decrease in autonomous and cofactor-dependent PKC activity likewise was prevented by okadaic acid. These findings suggest that LTD in the adult hippocampus in vivo involves a decrease in PKC activity that is mediated, at least in part, by dephosphorylation of the catalytic domain of PKC by protein phosphatases activated after LTD-inducing stimulation. Our findings are consistent with the idea that protein dephosphorylation contributes to the expression of LTD.

Impairment of long-term potentiation and associative memory in mice that overexpress extracellular superoxide dismutase.

Reactive oxygen species, including superoxide, generally are considered neurotoxic molecules whose effects can be alleviated by antioxidants. Different from this view, we show that scavenging of superoxide with an antioxidant enzyme is associated with deficits in hippocampal long-term potentiation (LTP), a putative neural substrate of memory, and hippocampal-mediated memory function. Using transgenic mice that overexpress extracellular superoxide dismutase (EC-SOD), a superoxide scavenger, we found that LTP was impaired in hippocampal area CA1 despite normal LTP in area CA3. The LTP impairment in area CA1 could be reversed by inhibition of EC-SOD. In addition, we found that EC-SOD transgenic mice exhibited impaired long-term memory of fear conditioning to contextual cues despite exhibiting normal short-term memory of the conditioning experience. These findings strongly suggest that superoxide, rather than being considered exclusively a neurotoxic molecule, should also be considered a signaling molecule necessary for normal neuronal function.

The extracellular signal-regulated kinase cascade is required for NMDA receptor-independent LTP in area CA1 but not area CA3 of the hippocampus.

Activation of extracellular signal-regulated kinase (ERK) has been shown to be necessary for NMDA receptor-dependent long-term potentiation (LTP). We studied the role of ERK in three forms of NMDA receptor-independent LTP: LTP induced by very high-frequency stimulation (200 Hz-LTP), LTP induced by the K(+) channel blocker tetraethylammonium (TEA) (TEA-LTP), and mossy fiber (MF) LTP (MF-LTP). We found that ERK was activated in area CA1 after the induction of both 200 Hz-LTP and TEA-LTP and that this activation required the influx of Ca(2+) through voltage-gated Ca(2+) channels. Inhibition of the ERK signaling cascade with either PD 098059 or U0126 prevented the induction of both 200 Hz-LTP and TEA-LTP in area CA1. In contrast, neither PD 098059 nor U0126 prevented MF-LTP in area CA3 induced by either brief or long trains of high-frequency stimulation. U0126 also did not prevent forskolin-induced potentiation in area CA3. However, incubation of slices with forskolin, an activator of the cAMP-dependent protein kinase (PKA) cascade, did result in increases in active ERK and cAMP response element-binding protein (CREB) phosphorylation in area CA3. The forskolin-induced increase in active ERK was inhibited by U0126, whereas the increase in CREB phosphorylation was not, which suggests that in area CA3 the PKA cascade is not coupled to CREB phosphorylation via ERK. Overall, our observations indicate that activation of the ERK signaling cascade is necessary for NMDA receptor-independent LTP in area CA1 but not in area CA3 and suggest a divergence in the signaling cascades underlying NMDA receptor-independent LTP in these hippocampal subregions.

Synapse-specific compartmentalization of signaling cascades for LTP induction in CA3 interneurons.

Inhibitory interneurons with somata in strata radiatum and lacunosum-molecular (SR/L-M) of hippocampal area CA3 receive excitatory input from pyramidal cells via the recurrent collaterals (RCs), and the dentate gyrus granule cells via the mossy fibers (MFs). Here we demonstrate that Hebbian long-term potentiation (LTP) at RC synapses on SR/L-M interneurons requires the concomitant activation of calcium-impermeable AMPARs (CI-AMPARs) and N-methyl-d-aspartate receptors (NMDARs). RC LTP was prevented by voltage clamping the postsynaptic cell during high-frequency stimulation (HFS; 3 trains of 100 pulses delivered at 100 Hz every 10s), with intracellular injections of the Ca(2+) chelator BAPTA (20mM), and with the NMDAR antagonist D-AP5. In separate experiments, RC and MF inputs converging onto the same interneuron were sequentially activated. We found that RC LTP induction was blocked by inhibitors of the calcium/calmodulin-dependent protein kinase II (CaMKII; KN-62, 10 µM or KN-93, 10 µM) but MF LTP was CaMKII independent. Conversely, the application of the protein kinase A (PKA) activators forskolin/IBMX (50 µM/25 µM) potentiated MF EPSPs but not RC EPSPs. Together these data indicate that the aspiny dendrites of SR/L-M interneurons compartmentalize synapse-specific Ca(2+) signaling required for LTP induction at RC and MF synapses. We also show that the two signal transduction cascades converge to activate a common effector, protein kinase C (PKC). Specifically, LTP at RC and MF synapses on the same SR/LM interneuron was blocked by postsynaptic injections of chelerythrine (10 µM). These data indicate that both forms of LTP share a common mechanism involving PKC-dependent signaling modulation.

Dendritic morphology and its effects on the amplitude and rise-time of synaptic signals in hippocampal CA3 pyramidal cells.

Detailed anatomical analysis and compartmental modeling techniques were used to study the impact of CA3b pyramidal cell dendritic morphology and hippocampal anatomy on the amplitude and time course of dendritic synaptic signals. We have used computer-aided tracing methods to obtain accurate three-dimensional representations of 8 CA3b pyramidal cells. The average total dendritic length was 6,332 +/- 1,029 microns and 5,062 +/- 1,397 microns for the apical and basilar arbors, respectively. These cells also exhibited a rough symmetry in their maximal transverse and septotemporal extents (311 +/- 84 microns and 269 +/- 106 microns). From the calculated volume of influence (the volume of the neuropil from which the dendritic structures can receive input), it was found that these cells show a limited symmetry between their proximal apical and basilar dendrites (2.1 +/- 1.2 x 10(6) microns 3 and 3.5 +/- 1.1 x 10(6) microns 3, respectively). Based upon these data, we propose that the geometry of these cells can be approximated by a combination of two cones for the apical arbor and a single cone for the basilar arbor. The reconstructed cells were used to build compartmental models and investigate the extent to which the cellular anatomy determines the efficiency with which dendritic synaptic signals are transferred to the soma. We found that slow, long lasting signals show only approximately a 50% attenuation when they occur in the most distal apical dendrites. However, synaptic transients similar to those seen in fast glutamatergic transmission are transferred much less efficiently, showing up to a 95% attenuation. The relationship between the distance along the dendrites and the observed attenuation for a transient is described simply by single exponential functions with parameters of 195 and 147 microns for the apical and basilar arbors respectively. In contrast, there is no simple relation that describes how a transient is attenuated with respect to these cells' stratified inputs. This lack of a simple relationship arises from the radial orientation of the proximal apical and basilar dendrites. When combined, the anatomical and modeling data suggest that a CA3b cell can be approximated in three dimensions as the combination of three cones. The amplitude and time-course for a synaptic transient can then be predicted using two simple equations.

Synaptic targets of the intrinsic axon collaterals of supragranular pyramidal neurons in monkey prefrontal cortex.

The principal axons of supragranular pyramidal neurons in the cerebral cortex travel through the white matter and terminate in other cortical areas, whereas their intrinsic axon collaterals course through the gray matter and form both local and long-distance connections within a cortical region. In the monkey prefrontal cortex (PFC), horizontally oriented, intrinsic axon collaterals from supragranular pyramidal neurons form a series of stripe-like clusters of axon terminals (Levitt et al. [1993] J Comp Neurol 338:360-376; Pucak et al. [1996] J Comp Neurol 376:614-630). The present study examined the synaptic targets of the intrinsic axon collaterals arising from supragranular pyramidal neurons within the same stripe (local projections). Approximately 50% of the within-stripe axon terminals in monkey PFC area 9 targeted dendritic spines. In contrast, for both the intrinsic axon collaterals that travel between stripes (long-range projections), and the axon terminals that project to other PFC areas (associational projections), over 92% of the postsynaptic structures were dendritic spines (Melchitzky et al. [1998] J Comp Neurol 390:211-224). The other 50% of the within-stripe terminals synapsed with dendritic shafts. Dual-labeling studies confirmed that these within-stripe terminals contacted gamma-aminobutyric acid-immunoreactive dendritic shafts, including the subpopulation that contains the calcium-binding protein parvalbumin. The functional significance of the differences in synaptic targets between local and long-range intrinsic axon collaterals was supported by whole-cell, patch clamp recordings in an in vitro slice preparation of monkey PFC. Specifically, the small amplitude responses observed in layer 3 pyramidal neurons during long-range, low-intensity stimulation were exclusively excitatory, whereas local stimulation also evoked di/polysynaptic inhibitory responses. These anatomic and electrophysiological findings suggest that intrinsic connections of the PFC differ from other cortical regions and that within the PFC, feedback (within-stripe) inhibition plays a greater role in regulating the activity of supragranular pyramidal neurons than does feedforward inhibition either between stripes or across regions.

Morphology of developing rat genioglossal motoneurons studied in vitro: changes in length, branching pattern, and spatial distribution of dendrites.

The aim of this study is to describe the postnatal change in dendritic morphology of those motoneurons in the hypoglossal nucleus that innervate the genioglossus muscle. Forty genioglossal (GG) motoneurons from four age groups (1-2, 5-6, 13-15, and 19-30 postnatal days) were labeled by intracellular injection of neurobiotin in an in vitro slice preparation of the rat brainstem and were reconstructed in three-dimensional space. The number of primary dendrites per GG motoneuron was approximately 6 and remained unchanged with age. The development of these motoneurons from birth to 13-15 days was characterized by a simplification of the dendritic tree involving a decrease in the number of terminal endings and dendritic branches. Motoneurons lost their 6th-8th order branches, in parallel with an elongation of their terminal dendritic branches maintaining the same combined dendritic length. The elongation of terminal branches was attributed to both longitudinal growth and the apparent lengthening caused by resorption of distal branches. The elimination of dendritic branches tended to increase the symmetry of the tree, as revealed by topological analysis. Later, between 13-15 days and 19-30 days, there was a reelaboration of the dendritic arborization returning to a configuration similar to that found in the newborn. The length of terminal branches was shorter at 19-30 days, while the length of preterminal branches did not change, suggesting that the proliferation of branches at 19-30 days takes place in the intermediate parts of terminal branches. The three-dimensional distribution of dendrites was analyzed by dividing space into six equal volumes (hexants). This analysis revealed that GG motoneurons have major components of their dendritic tree oriented in the lateral, medial, and dorsal hexants. Further two-dimensional polar analysis (consisting of eight sectors) revealed a reconfiguration of the tree from birth up to 5-6 days involving resorption of dendrites in the dorsal, dorsomedial, and medial sectors and growth in the lateral sector. Later in development (between 13-15 days and 19-30 days), there was growth in all sectors, but of a greater magnitude in the dorsomedial, medial, and dorsolateral sectors.

Heterosynaptic correlates of long-term potentiation induction in hippocampal CA3 neurons.

Previous studies have demonstrated that tetanization of hippocampal mossy fibers induces a long-term potentiation of non-tetanized (heterosynaptic) non-mossy fiber afferents (Schaffer collateral/commissural and fimbrial fibers). Tetanization of these non-mossy fiber afferents, in contrast, does not induce mossy fiber long-term potentiation, but induces a long-term depression of mossy fiber responses (Bradler and Barrionuevo, Synapse 4, 132-142, 1989). The synaptic activity necessary to evoke these heterosynaptic alterations of efficacy is not known. Specifically, the dependence of heterosynaptic efficacy on the activation of N-methyl-D-aspartate receptors has not been assessed. In addition, the capability of different afferents to CA3 neurons to support alterations in heterosynaptic efficacy remains largely unknown. In the present study, heterosynaptic alterations of efficacy in the rat did not require the activation of N-methyl-D-aspartate receptors. Mossy fibers supported N-methyl-D-aspartate receptor-independent heterosynaptic long-term depression, and N-methyl-D-aspartate receptor-independent long-term potentiation. In contrast, non-mossy fiber afferents expressed N-methyl-D-aspartate receptor-independent heterosynaptic long-term potentiation induced by a mossy fiber tetanus, and an N-methyl-D-aspartate receptor-independent long-term depression, in addition to N-methyl-D-aspartate receptor-dependent homosynaptic long-term potentiation. The possibility that non-N-methyl-D-aspartate receptor activity in non-tetanized afferents is necessary for heterosynaptic long-term potentiation induction is discussed. Heterosynaptic long-term depression was induced in the absence of homosynaptic long-term potentiation, suggesting that these concomitant forms of synaptic plasticity rely on different mechanisms.

The multifarious hippocampal mossy fiber pathway: a review.

The hippocampal mossy fiber pathway between the granule cells of the dentate gyrus and the pyramidal cells of area CA3 has been the target of numerous scientific studies. Initially, attention was focused on the mossy fiber to CA3 pyramidal cell synapse because it was suggested to be a model synapse for studying the basic properties of synaptic transmission in the CNS. However, the accumulated body of research suggests that the mossy fiber synapse is rather unique in that it has many distinct features not usually observed in cortical synapses. In this review, we have attempted to summarize the many unique features of this hippocampal pathway. We also have attempted to reconcile some discrepancies that exist in the literature concerning the pharmacology, physiology and plasticity of this pathway. In addition we also point out some of the experimental challenges that make electrophysiological study of this pathway so difficult.Finally, we suggest that understanding the functional role of the hippocampal mossy fiber pathway may lie in an appreciation of its variety of unique properties that make it a strong yet broadly modulated synaptic input to postsynaptic targets in the hilus of the dentate gyrus and area CA3 of the hippocampal formation.

Transient and persistent increases in protein phosphatase activity during long-term depression in the adult hippocampus in vivo.

The neural substrates of learning and memory most likely involve activity-dependent long-term changes in synaptic strength, including long-term potentiation and long-term depression. A critical element in the cascade of events hypothesized to underlie such changes in synaptic function is modification of protein phosphorylation. Long-term depression is thought to involve decreases in protein phosphorylation, which could result from reduction in protein kinase activity and/or enhancement in protein phosphatase activity. We present here direct evidence that long-term depression in the hippocampus in vivo is associated with an increase in the activity of the serine/threonine phosphatases 1 and 2A. The increase in activity of phosphatase 1 was transient, whereas that of phosphatase 2A lasted > 65 min after the induction of long-term depression. Blockade of long-term depression prevented the observed increases in phosphatase activity, as did selective inhibition of phosphatase 1 and 2A. Induction of long-term depression had no effect on the level of either phosphatase, which suggests that our results reflect increases in the intrinsic activity of these two enzymes. Our findings are consistent with a model of synaptic plasticity that implicates protein dephosphorylation by serine/threonine phosphatases in the early maintenance and/or expression of long-term depression of synaptic strength.

Release of D,L-threo-beta-hydroxyaspartate as a false transmitter from excitatory amino acid-releasing nerve terminals.

This study examined whether preaccumulated D,L-threo-beta-hydroxyaspartate (tHA), a competitive substrate for the high-affinity excitatory amino acid (EAA) transporter, is released as a false transmitter from EAA-releasing nerve terminals. Potassium-stimulation (50 mM for 1 min) evoked significant release of the endogenous EAAs (aspartate and glutamate) from superfused neocortical minislices. Endogenous EAA release was largely calcium-dependent and was inhibited by tetanus toxin, a neurotoxin which specifically blocks vesicular exocytosis. In parallel experiments, minislices were pre-incubated with 500 microM tHA. Potassium (50 mM) evoked significant release of tHA and this release was also calcium-dependent and reduced by tetanus toxin. Pre-accumulation of tHA did not affect the release of endogenous glutamate whereas the release of endogenous aspartate was significantly attenuated. These data suggest that tHA selectively accumulates in a vesicular aspartate pool and is released upon depolarization as a false transmitter from EAA nerve terminals.

Synaptosomal and vesicular accumulation of L-glutamate, L-aspartate and D-aspartate.

We examined the vesicular accumulation of the excitatory amino-acid (EAA) neurotransmitters, L-glutamate and L-aspartate, together with the non-metabolisable EAA analogue D-aspartate. Synaptosomes derived from whole brain were incubated in various concentrations of [3H]-amino acids under conditions to facilitate vesicular turnover. Synaptosomes were then lysed in hypotonic medium and vesicles immunoprecipitated with monoclonal anti-synaptophysin antibodies coupled to sepharose beads. Using this method, saturable vesicular accumulation was observed for [3H]-L-glutamate, [3H]-L-aspartate, and [3H]-D-aspartate but not for the excitatory amino acid receptor ligands [3H]-AMPA or [3H]-kainate. Vesicular accumulation (t(1/2)=7.45 min) was markedly slower than synaptosomal accumulation (t(1/2)=1.03 min) and was substantially reduced at 4 degrees C. Maximal accumulation of [3H]-L-glutamate, [3H]-L-aspartate, and [3H]-D-aspartate was estimated to be 98, 68, and 112 pmol/mg of synaptosomal protein, respectively, and uptake affinities 1.6, 3.4, and 2.1 mM, respectively. Maximal accumulation of [3H]-L-glutamate was non-competitively inhibited by both 100 microM unlabeled L-aspartate and 100 microM D-aspartate, suggesting that all are accumulated into a common vesicular pool by different transporters.

Potassium-induced long-term potentiation in rat hippocampal slices.

We observed that a transient increase in extracellular potassium concentration (50 mM for 40 s) was sufficient to induce long-term potentiation (LTP) of synaptic transmission in area CA1 of the hippocampal slice. Potassium-induced potentiation of the Schaffer collateral/commissural synapses demonstrated several features characteristic of tetanus-induced LTP: (1) population excitatory post-synaptic potential (EPSP) amplitudes were enhanced to a similar magnitude (on average 70% above baseline) which (2) lasted for more than 20 min; (3) induction was blocked by bath application of the specific N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino-5-phosphonovalerate (D-APV), and (4) was attenuated by reduction of the concentration of calcium in the extracellular medium. Induction of either potassium-induced LTP or tetanus-induced LTP occluded the subsequent expression of the other. Finally, exposure to high potassium in the absence of electrical stimulation was sufficient to induce LTP. Taken together, these data indicate that brief depolarizing stimuli other than tetanus can induce LTP. Because potassium-induced LTP is not restricted to the subset of afferents examined electrophysiologically, such a method could facilitate analyses of the biochemical events underlying both the induction and expression of LTP.

Neurophysiological changes in the in vitro rat hippocampus following chronic lithium administration.

The effect of chronic lithium exposure on the electrophysiological responses of the Schaffer collateral-commissural (SCC) input to the CA1 pyramidal neurons in the in vitro hippocampus was investigated. Experimental animals were intubated intragastrically with lithium carbonate (150 mg/kg) for 3-4 weeks. This treatment produced lithium levels in serum and hippocampus of 0.3-1.3 mM. During the recording period, the hippocampal slice retained a stable lithium concentration of 53% of the initial value. Chronic lithium exposure had a depressive effect on input/output relationships, paired-pulse facilitation and strength of orthodromic inhibition. The antidromic inhibition was virtually intact. No obvious differences were found between control and lithium slices in amplitude, latency or waveforms, of synaptic and antidromic extracellular potentials. These findings are compatible with a major action of lithium on the excitability of SCC axons and synaptic terminals.

Major differences between long-term potentiation and ACPD-induced slow onset potentiation in hippocampus.

We examined the effects of a long-lasting application of the selective metabotropic glutamate receptor (mGluR) agonist 1S-3R, 1-amino cyclopentane-1,3-dicarboxylic acid (ACPD) on synaptic potentials recorded from the CA1 and CA3 subfields in hippocampal slices maintained in a superfusion slice chamber. In 25% of the slices, ACPD generated an slow onset potentiation (SOP) of population EPSPs (pEPSPs) in CA1. In contrast to long-term potentiation (LTP) induced by a tetanic train, SOP was accompanied by an increase in the magnitude of the presynaptic fiber volley. Potentiation of the isolated afferent volley suggests that the expression of SOP is due to a recruitment of additional presynaptic fibers by the test stimulus caused by the persistent block of K+ channels by ACPD.

Vincamine: a psychogeriatric agent blocking synaptic potentiation in hippocampus.

The action of vincamine on the physiology of the CA1 region of the in vitro hippocampal slice preparation was investigated. At concentrations of 1, 10 and 100 microM, a five-minute perfusion with vincamine did not affect the synaptically-mediated activation of pyramidal neurons evoked by stimulation of the Schaffer-commissural fiber system. The effect of vincamine on the excitability of the pyramidal neurons was investigated by studying its effect on the antidromically-elicited field potential and the input-output relation of Schaffer-commissural fiber input. No effect on either of the two parameters was seen at a concentration of 100 microM of vincamine. Vincamine did, however, attenuate both the post-tetanic (PTP) and long-term potentiation (LTP) evoked by repetitive stimulation of the Schaffer-commissural fiber system. At a concentration of 100 microM of vincamine, PTP was significantly reduced and LTP was almost completely suppressed.

Dopamine modulation of neuronal function in the monkey prefrontal cortex.

We developed a brain slice preparation that allowed us to apply whole-cell recordings to examine the electrophysiological properties of identified synapses, neurons, and local circuits in the dorsolateral prefrontal cortex (DLPFC) of macaque monkeys. In this article, we summarize the results from some of our recent and current in vitro studies in the DLPFC with special emphasis on the modulatory effects of dopamine (DA) receptor activation on pyramidal and nonpyramidal cell function in superficial layers in DLPFC areas 46 and 9.

Afterdischarges elicited by electrical thalamic stimulation in the hemicerebellectomized baboon.

Electrical stimulation of the ventrolateral (VL) nucleus of the thalamus was performed on hemicerebellectomized baboons. Stimulation was delivered in volleys of 40 to 60 Hz lasting 5 to 25 sec, and at intensities ranging from 1 to 6 mA (width of shock 0.5 to 5 msec). Similar parameters of stimulation produced asymmetrical motor and electrocortical responses. When the normal VL nucleus was stimulated, these responses were tonic or tonic-clonic. When the VL nucleus deprived of its cerebellar afferents was stimulated, the tonic-clonic responses were followed by afterdischarges appearing in the EEG which persisted after cessation of stimulation. The role of the cerebellum in the lowering of the excitability threshold of some structures, particularly the VL nucleus, is discussed, as are the systems likely to play a role in producing various clinical responses.