Ephaptic feedback in identified synapses in mollusk neurons.

The possible existence of intrasynaptic ephaptic feedback in the invertebrate CNS was studied. Intracellular recordings were made of excitatory postsynaptic potentials and currents arising on activation of the recently described monosynaptic connection between identified neurons in the snail CNS. In the presence of ephaptic feedback, tetanization of the postsynaptic neuron with hyperpolarizing impulses should activate presynaptic calcium channels, thus increasing the amplitude of excitatory postsynaptic potential, while sufficiently strong postsynaptic hyperpolarization applied during generation of the excitatory postsynaptic current should induce "supralinear" increases in its amplitude, as has been observed previously in rat hippocampal neurons. The first series of experiments involved delivery of 10 trains of hyperpolarizing postsynaptic impulses (40-50 mV, duration 0.5 sec, frequency 1 Hz, train duration 45 sec); significant changes in the amplitude of excitatory postsynaptic were not seen. In the second series of experiments, changes in the amplitude of the excitatory postsynaptic current were studied during hyperpolarization of the postsynaptic neuron. At a potential of -100 mV, the amplitude of the excitatory postsynaptic current increased significantly more than predicted by its "classical" linear relationship with membrane potential. This "supralinear" increase in the amplitude of the excitatory postsynaptic potential can be explained by the operation of ephaptic feedback and is the first evidence for this phenomenon in CNS synapses of invertebrates.

[Ephaptic feedback in identified synapses of terrestrial snails]. / Efapticheskaia obratnaia sviaz' v identsifitsirovannom sinapse nazemnogo molliuska.

A hypothesis for the existence of the intrasynaptic ephaptic feedback (EFB) in the invertebrate central nervous sytem was tested. Excitatory postsynaptic potentials (EPSPs) and currents (EPSCs) evoked by the activation of the recently described monosynaptic connection between the identified snail neurons were recorded intracellularly. In case of the EFB presence, the postsynaptic tetanization with hyperpolarization pulses could activate presynaptic Ca2+ channels and enhance the EPSP amplitude, whereas a steady postsynaptic hyperpolarization should induce a "supralinear" increase in EPSC amplitudes as it has been found in the rat hippocampus. In the first series of the experiments, 10 trains of hyperpolarizing pulses (40-50 mV, 1 Hz, pulse duration 0.5 s, train duration 45 s) were delivered postsynaptically. No significant changes in EPSP amplitudes were found. In the second series of the experiments, the EPSC amplitudes were measured during varying postsynaptic hyperpolarization. At the membrane potential 100 mV, the EPSP amplitude was significantly higher than theoretically predicted from the classical linear dependence. Such a "supralinear" effect of postsynaptic depolarization can be explained by the presence of the EFB. This finding is the first evidence for the EFB existence in the invertebrate central nervous system.

Postsynaptic depolarisation enhances transmitter release and causes the appearance of responses at "silent" synapses in rat hippocampus.

Recent data indicate that most "silent" synapses in the hippocampus are "presynaptically silent" due to low transmitter release rather than "postsynaptically silent" due to "latent" receptors of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid type (AMPARs). That synapses bearing only N-methyl-d-aspartate (NMDAR) receptors do exist is suggested by the decreased number of transmission failures during postsynaptic depolarisation and by the presence of NMDA-mediated excitatory postsynaptic currents (EPSCs) in synapses silent at rest. We tested whether these effects could be due to potentiated transmitter release at depolarised postsynaptic potentials rather than removal of Mg(2+) block from NMDARs. Using whole-cell recordings of minimal EPSCs from CA1 and CA3 neurones of hippocampal slices we confirmed decreased incidence of failures at +40 mV as compared with -60 mV. This effect was associated with a gradual increase of EPSC amplitude after switching to +40 mV and with a decrease of paired-pulse facilitation. In initially silent synapses, potentiation of pharmacologically isolated AMPAR-mediated EPSCs was still observed at +40 mV and this persisted after stepping back to -60 mV. All above effects were blocked when the cell was dialysed with the Ca(2+) chelator BAPTA (20 mM). These observations are difficult to reconcile with the "latent AMPAR" hypothesis and suggest an alternative explanation, namely that the reduction in failure rates at positive potentials is due to potentiation of transmitter release following Ca(2+) influx through NMDARs. Our results suggest that silent synapses can be mainly "presynaptically" rather than "postsynaptically silent" and thus increased transmitter release rather than insertion of AMPARs is a major mechanism of early long-term potentiation maintenance.

Stimulation of the lateral hypothalamus provokes the initiation of robust long-term potentiation of the thalamo-cortical input to the barrel field of the adult, freely moving rat.

Long-term potentiation in the thalamo-cortical input to the somatosensory cortex barrel field has been reported to be inducible in vitro only during a narrow critical period of the first postnatal week. Here we explored whether this is due to inability of adult synapses to express LTP or lack of appropriate conditions for LTP induction in slice preparations. We recorded thalamo-cortical field potentials (FPs) from the barrel field of chronically prepared adult rats. In the first series, several parameters of conditioning tetanization of thalamus (T) have been tried. Statistically significant LTP of 135-150% relative to the baseline was observed only in rare cases (3/18) so that the mean changes were not statistically significant. In the second series, five trains of 100 Hz stimulation of T were paired with a "reinforcing" stimulation of the lateral hypothalamus (LH). In most cases (9/13), thalamo-cortical FPs were potentiated. The mean post-tetanic amplitude was 238 +/- 42% (+/- SEM) relative to the baseline (n = 13). The potentiation persisted for >1 h and typically even further increased when tested 24-48 h later. LTP magnitude strongly correlated with the initial paired-pulse ratio (PPR, coefficient of correlation r = 0.98) so that the LTP magnitude was larger (333 +/- 107, n = 6) in cases with PPR > 1.3. The mean PPR tended to decrease after LTP (from 2.05 to 1.65). Altogether the results suggest that LTP is inducible in the thalamo-cortical input to the barrel field of normal adult rats. The dependence of the LTP magnitude upon the initial PPR suggests that inputs with low initial release probability undergo larger LTP. Together with the tendency to a decrease in the PPR this suggests an involvement of presynaptic mechanisms in the maintenance of neocortical LTP.

Stimulation of the lateral hypothalamus provokes the initiation of robust long-term potentiation of the thalamo-cortical input to the barrel field of the adult, freely moving rat.

Long-term potentiation in the thalamo-cortical input to the somatosensory cortex barrel field has been reported to be inducible in vitro only during a narrow critical period of the first postnatal week. Here we explored whether this is due to inability of adult synapses to express LTP or lack of appropriate conditions for LTP induction in slice preparations. We recorded thalamo-cortical field potentials (FPs) from the barrel field of chronically prepared adult rats. In the first series, several parameters of conditioning tetanization of thalamus (T) have been tried. Statistically significant LTP of 135-150% relative to the baseline was observed only in rare cases (3/18) so that the mean changes were not statistically significant. In the second series, five trains of 100 Hz stimulation of T were paired with a "reinforcing" stimulation of the lateral hypothalamus (LH). In most cases (9/13) thalamo-cortical FPs were potentiated. The mean post-tetanic amplitude was 238 +/- 42% (+/- SEM) relative to the baseline (n = 13). The potentiation persisted for > > 1 hr and typically even further increased when tested 24-48 hr later. LTP magnitude strongly correlated with the initial paired-pulse ratio (PPR, coefficient of correlation r = 0.98) so that LTP magnitude was larger (333 +/- 107, n = 6) in cases with PPR > 1.3. The mean PPR tended to decrease after LTP (from 2.05 to 1.65). Altogether the results suggest that LTP is inducible in the thalamo-cortical input to the barrel field of normal adult rats. The dependence of LTP magnitude upon the initial PPR suggests that inputs with low initial release probability undergo larger LTP. Together with the tendency to a decrease in the PPR this suggests an involvement of presynaptic mechanisms in the maintenance of neocortical LTP.

Low-frequency depression of synaptic responses recorded from rat visual cortex.

To characterize the low-frequency depression (LFD) of synaptic transmission in the visual cortex, we recorded field potentials and minimal excitatory postsynaptic potentials (EPSPs) from layer II/III following intracortical stimulation at various frequencies in cortical slices of rats. Field potentials were stable at 0.017 Hz, but showed an amplitude depression at 0.033-0.1 Hz at stimulus intensity of 1.5 times the threshold for induction of the postsynaptic component and at 0.1-0.2 Hz at intensity of 1.2 times the threshold. The LFD was input-specific and its magnitude correlated with the stimulus frequency. An interruption of stimulation for 15 min yielded a nearly complete recovery from LFD. Minimal EPSPs tested at 0.1-1.7 Hz often showed LFD with similar features. However, some inputs were stable or even facilitated during repeated stimulation. At 0.1 and 0.2 Hz, >50% of inputs were stable, whereas 10% and 25% were depressed, respectively. At 0.5 and 1.7 Hz, LFD was observed in >60% and 80% of inputs, respectively. The magnitude of LFD strongly varied across inputs. In 3 of the 41 inputs analyzed, LFD was so strong that these inputs became virtually silent. Occurrence of responses to the second pulse in the paired-pulse paradigm when the first response was absent and recovery of depressed EPSPs following stimulus interruption or shift to a lower frequency suggest that these synapses were presynaptically silent due to a lowered probability of transmitter release. Altogether, the results indicate that testing intervals of <10 or even < or =30 s cannot be regarded as completely neutral. At the single-cell level, frequency-dependent changes were strongly heterogeneous across different inputs. LFD and its spontaneous recovery may underlie the previously described "post-rest" potentiation, and should be taken into account when considering information processing in cortical networks.

Quantal analysis suggests strong involvement of presynaptic mechanisms during the initial 3 h maintenance of long-term potentiation in rat hippocampal CA1 area in vitro.

Long-term potentiation (LTP) is the most prominent model to study neuronal plasticity. Previous studies using quantal analysis of an early stage of LTP in the CA1 hippocampal region (<1 h after induction) suggested increases in both the mean number of transmitter quanta released by each presynaptic pulse (m, quantal content) and postsynaptic effect of a single quantum (v, quantal size). When LTP was large, it was m that increased predominantly suggesting prevailing presynaptic contribution. However, LTP consists of several temporary phases with presumably different mechanisms. Here we recorded excitatory postsynaptic potentials from CA1 hippocampal slices before and up to 3.5 h after LTP induction. A new version of the noise deconvolution revealed significant increases in m with smaller and often not statistically significant changes in v. The changes in m were similar for both early (<1 h) and later (1-3 h) post-tetanic periods and correlated with LTP magnitude. The coefficient of variation of the response amplitude and the number of failures decreased during both early and late post-tetanic periods. The results suggest that both early (<0.5 h) and later LTP components (0.5-3 h) are maintained by presynaptic changes, which include increases in release probabilities and the number of effective release sites. In addition initially silent synapses can be converted into effective ones due to either pre- or postsynaptic rearrangements. If this occurs, our data indicate that the number and the efficacy of the receptors in the new transmission sites are approximately similar to those in the previously effective sites.

Long-term potentiation of the AMPA and NMDA components of minimal postsynaptic currents in rat hippocampal field Ca1.

The aim of the present work was to study the potentiation of the AMPA and NMDA components of minimal excitatory postsynaptic currents (EPSC) evoked by activation of restricted numbers of synapses. EPSC of neurons in field CA1 in hippocampal slices were recorded in whole-call patch-clamp conditions selected such that both (AMPA and NMDA) components were present, and these were measured in parallel using computational methods in combination with pharmacological receptor blockade. There was a quite strong correlation between the amplitudes of the AMPA and NMDA components and this was regarded as evidence that they were generated by the same synapses. In cases producing this correlation, both components showed essentially equal long-term potentiation lasting from 5 min to 2 h after afferent tetanization. The data did not support the postsynaptic hypothesis and were in better agreement with the concept that the major mechanism for the persistence of the initial phase of long-term potentiation (up to 1-2 h) is based on increases in the quantity of transmitter released presynaptically.

Presynaptic R-type calcium channels contribute to fast excitatory synaptic transmission in the rat hippocampus.

The possibility that R-type calcium channels contribute to fast glutamatergic transmission in the hippocampus has been assessed using low concentrations of NiCl(2) and the peptide toxin SNX 482, a selective antagonist of the pore-forming alpha(1E) subunit of R-type calcium channel. EPSPs or EPSCs were recorded in the whole-cell configuration of the patch-clamp technique mainly from CA3 hippocampal neurons. Effects of both NiCl(2) and SNX 482 were tested on large (composite) EPSCs evoked by mossy and associative-commissural fiber stimulation. NiCl(2) effects were also tested on minimal EPSPs-EPSCs. Both substances reduced the amplitude of EPSPs-EPSCs. This effect was associated with an increase in the number of response failures of minimal EPSPs-EPSCs, an enhancement of the paired-pulse facilitation ratios of both minimal and composite EPSCs, and a reduction of the inverse squared coefficient of variation (CV(-2)). The reduction of CV(-2) was positively correlated with the decrease in EPSC amplitude. The inhibitory effect of NiCl(2) was occluded by SNX 482 but not by omega-conotoxin-MVIIC, a broad-spectrum antagonist thought to interact with N- and P/Q-type calcium channels, supporting a specific action of low concentrations of NiCl(2) on R-type calcium channels. Together, these observations indicate that both NiCl(2) and SNX 482 act at presynaptic sites and block R-type calcium channels with pharmacological properties similar to those encoded by the alpha(1E) gene. These channels are involved in fast glutamatergic transmission at hippocampal synapses.

[Long-term potentiation of AMPA- and NMDA-components of minimal postsynaptic currents in CA1 area of the rat hippocampus]. / Dolgovremennaia potentsiatsiia AMPA- i NMDA-komponent minimal'nykh postsinapticheskikh tokov v oblasti CA1 gippokampa krys.

Excitatory postsynaptic currents (EPSC) were recorded from pyramidal neurons of the rat hippocampal slices as well as the AMPA and NMDA components. A strong enough correlation between the amplitudes of the components provided a reliable evidence of their generation by the same synapse. Both components revealed similar LTP following afferent tetanisation. The data obtained do not support postsynaptic mechanisms of the LTP maintenance but suggest that increased presynaptic release represents a basic mechanism of the early LTP maintenance.

Differences in amplitude-voltage relations between minimal and composite mossy fibre responses of rat CA3 hippocampal neurons support the existence of intrasynaptic ephaptic feedback in large synapses.

Computer simulations and electrophysiological experiments have been performed to test the hypothesis on the existence of an ephaptic interaction in purely chemical synapses. According to this hypothesis, the excitatory postsynaptic current would depolarize the presynaptic release site and further increase transmitter release, thus creating an intrasynaptic positive feedback. For synapses with the ephaptic feedback, computer simulations predicted non-linear amplitude-voltage relations and voltage dependence of paired-pulse facilitation. The deviation from linearity depended on the strength of the feedback determined by the value of the synaptic cleft resistance. The simulations showed that, in the presence of the intrasynaptic feedback, recruitment of imperfectly clamped synapses and synapses with linear amplitude-voltage relations tended to reduce the non-linearity and voltage dependence of paired-pulse facilitation. Therefore, the simulations predicted that the intrasynaptic feedback would particularly affect small excitatory postsynaptic currents induced by activation of electrotonically close synapses with long synaptic clefts. In electrophysiological experiments performed on hippocampal slices, the whole-cell configuration of the patch-clamp technique was used to record excitatory postsynaptic currents evoked in CA3 pyramidal cells by activation of large mossy fibre synapses. In accordance with the simulation results, minimal excitatory postsynaptic currents exhibited "supralinear" amplitude-voltage relations at hyperpolarized membrane potentials, decreases in the failure rate and voltage-dependent paired-pulse facilitation. Composite excitatory postsynaptic currents evoked by activation of a large amount of presynaptic fibres typically bear linear amplitude-voltage relationships and voltage-independent paired-pulse facilitation. These data are consistent with the hypothesis on a strong ephaptic feedback in large mossy fibre synapses. The feedback would provide a mechanism whereby signals from large synapses would be amplified. The ephaptic feedback would be more effective on synapses activated in isolation or together with electrotonically remote inputs. During synchronous activation of a large number of neighbouring inputs, suppression of the positive intrasynaptic feedback would prevent abnormal boosting of potent signals.

Intrasynaptic ephaptic feedback in central synapses.

Electrophysiological laboratory studies on rat visual cortex and hippocampus slices are reviewed. The aim was to confirm the existence of positive feedback in central synapses operating by an electrical (ephaptic) mechanism, as suggested by Byzov. Byzov's hypothesis holds that artificial hyperpolarization of the postsynaptic membrane potential should increase the amplitude of the excitatory postsynaptic current (EPSC) and potential (EPSP) in some central synapses not only by means of increases in the electromotive force (EMF). but also by means of increases in transmitter release from the presynaptic apparatus. Some experiments showed that hyperpolarization altered the parameters of presynaptic transmitter release, i.e., the quantity of "failed" responses N0, the coefficient of variation CV, and the quantum composition m of minimal EPSC and EPSP. The effect was particularly marked for EPSP in giant synapses formed by mossy fibers on neurons in field CA3. "Supralinear" functions were observed for these synapses in the relationship between EPSC amplitude and membrane potential in conditions of hyperpolarization of membrane potentials and in the relationship between presynaptic paired-stimulus facilitation and membrane potential. All of these "non-classical" effects disappeared when summed rather than minimal EPSC were evoked. The results are in agreement with computer experiments based on the Byzov model and are regarded as support for Byzov's hypothesis. Regardless of their explanation, the data obtained here demonstrate a new feedback mechanism for central synapses, which allows the postsynaptic neuron to control the efficiency of some synapses via changes in membrane potential. This mechanism can significantly increase the efficiency of large ("perforated") synapses and explains the increase in the number of this type of synapse after various experimental manipulations, such as those inducing long-term potentiation or forming conditioned reflexes.

Postsynaptic hyperpolarization increases the strength of AMPA-mediated synaptic transmission at large synapses between mossy fibers and CA3 pyramidal cells.

In chemical synapses information flow is polarized. However, the postsynaptic cells can affect transmitter release via retrograde chemical signaling. Here we explored the hypothesis that, in large synapses, having large synaptic cleft resistance, transmitter release can be enhanced by electrical (ephaptic) signaling due to depolarization of the presynaptic release site induced by the excitatory postsynaptic current itself. The hypothesis predicts that, in such synapses, postsynaptic hyperpolarization would increase response amplitudes "supralinearly", i.e. stronger than predicted from the driving force shift. We found supralinear increases in the amplitude of minimal excitatory postsynaptic potential (EPSP) during hyperpolarization of CA3 pyramidal neurons. Failure rate, paired-pulse facilitation, coefficient of variation of the EPSP amplitude and EPSP quantal content were also modified. The effects were especially strong on mossy fiber EPSPs (MF-EPSPs) mediated by the activation of large synapses and identified pharmacologically or by their kinetics. The effects were weaker on commissural fiber EPSPs mediated by smaller and more remote synapses. Even spontaneous membrane potential fluctuations were associated with supralinear MF-EPSP increases and failure rate reduction. The results suggest the existence of a novel mechanism for retrograde control of synaptic efficacy from postsynaptic membrane potential and are consistent with the ephaptic feedback hypothesis.

Silent synapses in the developing hippocampus: lack of functional AMPA receptors or low probability of glutamate release?

At early developmental stages, silent synapses have been commonly found in different brain areas. These synapses are called silent because they do not respond at rest but are functional at positive membrane potentials. A widely accepted interpretation is that N-methyl-d-aspartate (NMDA) but not alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors are functionally expressed on the subsynaptic membrane. Here we show that, in both CA3 and CA1 hippocampal regions, AMPA-mediated synaptic responses can be detected already at early stages of postnatal development. However, some synapses appear silent because of a very low probability of glutamate release. They can be converted into functional ones by factors that enhance release probability such as paired-pulse stimulation, increasing the temperature or cyclothiazide (CTZ), a drug that blocks AMPA receptor desensitization and increases transmitter release. Conversely, conducting synapses can be switched off by increasing the frequency of stimulation. Although we cannot exclude that "latent AMPA receptors" can become functional after activity-dependent processes, our results clearly indicate that, in the neonatal hippocampus, a proportion of glutamatergic synaptic connections are presynaptically rather than postsynaptically silent.

[Intrasynaptic ephaptic feedback in central synapses]. / Vnutrisinapticheskaia éfapticheskaia obratnaia sviaz' v tsentral'nykh sinapsakh.

A review is given of experiments performed in the author's laboratory on slices from the rat visual cortex and hippocampus. The aim was to test the existence of the positive feedback in central synapses according to a mechanism of electrical (ephatic) linking proposed by A. L. Byzow. The hypothesis predicts that, in a subset of central synapses, artificial postsynaptic membrane potential (MP) hyperpolarization should increase the amplitude of the excitatory postsynaptic current (EPSC) and potential (EPSP) not only due to a deviation from the equilibrium potential but also due to increased presynaptic transmitter release. In a part of the experiments, we found changes in several traditional parameters of transmitter release during hyperpolarization: number of response failures, coefficient of variation of response amplitude and quantal content of minimal EPSC/EPSP. The effects were especially prominent for the giant mossy fibre-CA3 synapses. For them, "supralinear" amplitude-voltage relations at hyperpolarized membrane potentials and voltage--dependent paired--pulse facilitation ratios were found. All these "non-classical" effects disappeared when composite, rather than minimal, EPSCs were evoked. These data were consistent with simulation experiments performed on the Byzov's synaptic model with the ephaptic feedback and therefore they strengthen the hypothesis. Independent of their interpretation, the data reveal a novel feedback mechanism. The mechanism provides a possibility for the central postsynaptic neurone to control the efficacy of a subset of synapses via postsynaptic MP modifications. The mechanism can essentially increase the efficacy of large ("perforated") synapses. It explains the significance of the increased number of such synapses following experimental challenges such as leading to induction of the long-term potentiation or to behavioural conditioning.

Intracellular studies of the interaction between paired-pulse facilitation and the delayed phase of long-term potentiation in the hippocampus.

The mechanisms of the early (up to 1 h) and late (up to 3 h) phases of long-term potentiation were studied by analyzing the interaction between long-term potentiation and presynaptic paired-pulse facilitation. "Minimal" excitatory postsynaptic potentials were measured in pyramidal neurons in field CA1 of rat hippocampal slices in conditions of paired-pulse stimulation of the radial layer. In most neurons, paired-pulse facilitation decreased after induction of long-term potentiation, and this reduction lasted throughout the recording period (up to 3.5 h). Changes in paired-pulse facilitation correlated with the extent of long-term facilitation and with the initial level of paired-pulse facilitation, and the extent of facilitation depended on the initial level of paired-pulse facilitation. This latter relationship was different for the early and late phases of long-term potentiation and increased with time. Overall, the data obtained here demonstrate a significant role for presynaptic mechanisms in maintaining both the early and late phases of long-term potentiation. It is suggested that the basic mechanism of the early phase of potentiation is an increase in the probability that transmitter will be released, which also leads to an increase in the number of effective release sites, due to transformation of "presynaptically quiet" synapses into effective synapses. It is proposed that the development of the late phase is based on simultaneous pre- and postsynaptic structural transformations which increase the number of synaptically active zones.

Long-term synaptic changes induced by intracellular tetanization of CA3 pyramidal neurons in hippocampal slices from juvenile rats.

Minimal excitatory postsynaptic potentials were evoked in CA3 pyramidal neurons by activation of the mossy fibres in hippocampal slices from seven- to 16-day-old rats. Conditioning intracellular depolarizing pulses were delivered as 50- or 100-Hz bursts. A statistically significant depression and potentiation was induced in four and five of 13 cases, respectively. The initial state of the synapses influenced the effect: the amplitude changes correlated with the pretetanic paired-pulse facilitation ratio. Afferent (mossy fibre) tetanization produced a significant depression in four of six inputs, and no significant changes in two inputs. Quantal content decreased or increased following induction of the depression or potentiation, respectively, whereas no significant changes in quantal size were observed. Compatible with presynaptic maintenance mechanisms of both depression and potentiation, changes in the mean quantal content were associated with modifications in the paired-pulse facilitation ratios, coefficient of variation of response amplitudes and number of response failures. Cases were encountered when apparently "presynaptically silent" synapses were converted into functional synapses during potentiation or when effective synapses became "presynaptically silent" when depression was induced, suggesting respective changes in the probability of transmitter release. It is concluded that, in juvenile rats, it is possible to induce lasting potentiation at the mossy fibre-CA3 synapses by purely postsynaptic stimulation, while afferent tetanization is accompanied by long-lasting depression. The data support the existence not only of a presynaptically induced, but also a postsynaptically induced form of long-term potentiation in the mossy fibre-CA3 synapse. Despite a postsynaptic induction mechanism, maintenance of both potentiation and depression is likely to occur presynaptically.