Postsynaptic factors control the duration of synaptic enhancement in area CA1 of the hippocampus.

In area of CA1 of the hippocampus, at least two phases of long-term potentiation (LTP) can be isolated: an early decremental component referred to as short-term potentiation (STP), which precedes a long-lasting, nondecremental component commonly considered to be stable LTP. Utilizing the hippocampal slice preparation, experiments were performed to determine the physiological factors controlling the conversion of STP to LTP. The duration of NMDA receptor-dependent synaptic enhancement was influenced by several factors, including the degree of postsynaptic NMDA receptor activation and the magnitude and timing of postsynaptic membrane depolarization during synaptic transmission. It was possible to convert STP to LTP by manipulations that increased the influx of calcium into the postsynaptic cell. These results demonstrate that NMDA receptor activation can result in distinct forms of synaptic potentiation and imply that the magnitude of postsynaptic calcium increase is a critical variable controlling the duration of synaptic enhancement.     

Dynamic action of neurometals at the synapse

There are synaptic vesicles that are labeled by Timm's sulfide-silver staining method in the brain, suggesting that synaptic vesicles contain metals such as zinc and copper. Zinc is co-released with glutamate and the importance of zinc signaling in the intracellular compartment, in addition to extracellular compartment, is becoming recognized. Zinc can pass through calcium channels, while blocking them. Calcium signaling plays a critical role for synaptic activity and crosstalk between zinc signaling with calcium signaling through calcium channels may participate in synaptic neurotransmission including synaptic plasticity such as long-term potentiation. Copper released into the synaptic cleft during synaptic excitation may also participate in synaptic neurotransmission. Other metals including copper potentially serve as calcium channel blockers and also influence calcium signaling and zinc signaling via the interaction with metal-binding proteins such as metallothioneins. Homeostasis of metals needs to be controlled spatiotemporally for proper brain function, and their dyshomeostasis is associated with neurological diseases. However, the data on the dynamic action of metals at synapses is limited and their significance poorly understood. This paper summarizes the action of metals in synaptic neurotransmission focused on calcium signaling at glutamatergic synapses.     

Biochemical effects of high-frequency synaptic activity studied with in vitro slices

Brain slices have a number of features that may be of value in the analysis of how physiological events affect neuronal chemistry. This paper discusses this topic and describes slice experiments concerned with the chemical events responsible for long-term potentiation (LTP) of synaptic responses found in hippocampus after brief episodes of high-frequency stimulation. Work with two variants of the slice procedure indicated that LTP is accompanied by an increase in the sodium-independent binding of [3H]glutamate to partially purified synaptic membranes; this effect very likely results from an increase in the numbers of a particular postsynaptic receptor. Stimulation that produces long-term potentiation also causes a significant change in the endogenous phosphorylation of pyruvate dehydrogenase (PDH), a key mitochondrial enzyme. Inasmuch as the phosphorylated state of PDH is strongly correlated with calcium sequestration by mitochondria, it is possible that LTP is triggered by a transient perturbation of the calcium buffering function provided by mitochondria. Low micromolecular levels of calcium increase glutamate binding to purified membranes apparently via the activation of a calcium-sensitive thiol proteinase. This mechanism could account for the increase in glutamate binding found in slices exhibiting LTP. These experiments suggest a possible explanation for long-term potentiation and indicate that slices can be used to detect at least some of the biochemical consequences of repetitive synaptic activity.     

The role of postsynaptic calcium in the induction of long-term potentiation

Long-term potentiation (LTP), a long-lasting, activity-dependent increase in the strength of synaptic transmission, is one of the most intensively studied forms of synaptic plasticity in the mammalian brain. In the CA1 region of the hippocampus, the induction of LTP is likely to require a rise in postsynaptic calcium levels. The main source for this calcium is influx through the NMDA receptor ionophore, although other potential sources include voltage-dependent calcium channels and release from intracellular stores. Dendritic spines, the sites of synaptic contact, may function to isolate and amplify synaptically mediated increases in postsynaptic calcium. Recent evidence indicates that the magnitude of postsynaptic calcium increase is a critical variable controlling the duration of synaptic enhancement. Although a number of calcium-dependent biochemical processes have been implicated in LTP, determining their exact role remains a challenging experimental problem.     

Calcium-Dependent Calcium Decay Explains STDP in a Dynamic Model of Hippocampal Synapses

It is widely accepted that the direction and magnitude of synaptic plasticity depends on post-synaptic calcium flux, where high levels of calcium lead to long-term potentiation and moderate levels lead to long-term depression. At synapses onto neurons in region CA1 of the hippocampus (and many other synapses), NMDA receptors provide the relevant source of calcium. In this regard, post-synaptic calcium captures the coincidence of pre- and post-synaptic activity, due to the blockage of these receptors at low voltage. Previous studies show that under spike timing dependent plasticity (STDP) protocols, potentiation at CA1 synapses requires post-synaptic bursting and an inter-pairing frequency in the range of the hippocampal theta rhythm. We hypothesize that these requirements reflect the saturation of the mechanisms of calcium extrusion from the post-synaptic spine. We test this hypothesis with a minimal model of NMDA receptor-dependent plasticity, simulating slow extrusion with a calcium-dependent calcium time constant. In simulations of STDP experiments, the model accounts for latency-dependent depression with either post-synaptic bursting or theta-frequency pairing (or neither) and accounts for latency-dependent potentiation when both of these requirements are met. The model makes testable predictions for STDP experiments and our simple implementation is tractable at the network level, demonstrating associative learning in a biophysical network model with realistic synaptic dynamics.     

BDNF acutely modulates synaptic transmission and calcium signalling in developing cortical neurons.

Brain-derived neurotrophic factor (BDNF), like other neurotrophins, has long-term effects on neuronal survival and differentiation; furthermore, BDNF has been reported to exert an acute potentiation of synaptic activity and are critically involved in long-term potentiation(LTP). We found that BDNF rapidly induced potentiation of synaptic activity and an increase in the intracellular Ca2+ concentration in cultured cortical neurons. Within minutes of BDNF application to cultured cortical neurons, spontaneous firing rate was dramatically increased as was the frequency and amplitude of excitatory spontaneous postsynaptic currents (EPSCs). Fura-2 recordings showed that BDNF acutely elicited an increase in intracellular calcium concentration ([Ca2+]i). This effect was partially dependent on extracellular Ca2+. In calcium-free perfusion medium a substantial calcium signal remained which disappeared after loading of cortical neurons with 5 microM U-73122. BDNF-induce Ca2+ transients were completely blocked by K252a and partially blocked by Cd2+. The results demonstrate that BDNF can enhance synaptic transmission and induce directly a rise in [Ca2+]i that require two routes: the release of Ca2+ from intracellular calcium stores and influx of extracellular Ca2+ mainly through voltage-dependent Ca2+ channels in cultured cortical neurons.     

Spatially restricted actions of BDNF

In this issue of Neuron, Zhang and Poo present evidence for localized BDNF-induced synaptic potentiation that is accompanied by spatially restricted calcium influx and requires local axonal protein synthesis. These results are consistent with a synapse-specific role for BDNF and provide a potentially novel way to think about cellular mechanisms for potentiation of neurotransmitter release.     

Endocannabinoids potentiate synaptic transmission through stimulation of astrocytes

Endocannabinoids and their receptor CB1 play key roles in brain function. Astrocytes express CB1Rs that are activated by endocannabinoids released by neurons. However, the consequences of the endocannabinoid-mediated neuron-astrocyte signaling on synaptic transmission are unknown. We show that endocannabinoids released by hippocampal pyramidal neurons increase the probability of transmitter release at CA3-CA1 synapses. This synaptic potentiation is due to CB1R-induced Ca(2+) elevations in astrocytes, which stimulate the release of glutamate that activates presynaptic metabotropic glutamate receptors. While endocannabinoids induce synaptic depression in the stimulated neuron by direct activation of presynaptic CB1Rs, they indirectly lead to synaptic potentiation in relatively more distant neurons by activation of CB1Rs in astrocytes. Hence, astrocyte calcium signal evoked by endogenous stimuli (neuron-released endocannabinoids) modulates synaptic transmission. Therefore, astrocytes respond to endocannabinoids that then potentiate synaptic transmission, indicating that astrocytes are actively involved in brain physiology.     

Dendritic Spike Saturation of Endogenous Calcium Buffer and Induction of Postsynaptic Cerebellar LTP

The architecture of parallel fiber axons contacting cerebellar Purkinje neurons retains spatial information over long distances. Parallel fiber synapses can trigger local dendritic calcium spikes, but whether and how this calcium signal leads to plastic changes that decode the parallel fiber input organization is unknown. By combining voltage and calcium imaging, we show that calcium signals, elicited by parallel fiber stimulation and mediated by voltage-gated calcium channels, increase non-linearly during high-frequency bursts of electrically constant calcium spikes, because they locally and transiently saturate the endogenous buffer. We demonstrate that these non-linear calcium signals, independently of NMDA or metabotropic glutamate receptor activation, can induce parallel fiber long-term potentiation. Two-photon imaging in coronal slices revealed that calcium signals inducing long-term potentiation can be observed by stimulating either the parallel fiber or the ascending fiber pathway. We propose that local dendritic calcium spikes, evoked by synaptic potentials, provide a unique mechanism to spatially decode parallel fiber signals into cerebellar circuitry changes.     

Single-cell optogenetic excitation drives homeostatic synaptic depression

Homeostatic processes have been proposed to explain the discrepancy between the dynamics of synaptic plasticity and the stability of brain function. Forms of synaptic plasticity such as long-term potentiation alter synaptic activity in a synapse- and cell-specific fashion. Although network-wide excitation triggers compensatory homeostatic changes, it is unknown whether neurons initiate homeostatic synaptic changes in response to cell-autonomous increases in excitation. Here we employ optogenetic tools to cell-autonomously excite CA1 pyramidal neurons and find that a compensatory postsynaptic depression of both AMPAR and NMDAR function results. Elevated calcium influx through L-type calcium channels leads to activation of a pathway involving CaM kinase kinase and CaM kinase 4 that induces synaptic depression of AMPAR and NMDAR responses. The synaptic depression of AMPARs but not of NMDARs requires protein synthesis and the GluA2 AMPAR subunit, indicating that downstream of CaM kinase activation divergent pathways regulate homeostatic AMPAR and NMDAR depression.     

The role of nitric oxide in hippocampal long-term potentiation.

Long-term potentiation is a long-lasting, use-dependent increase in the strength of synaptic connections. We investigated the role of nitric oxide (NO) in determining the duration of potentiation induced by high frequency stimulation of afferents in the CA1 region of the rat hippocampus. The calcium/calmodulin-dependent production of NO can be initiated by activation of excitatory amino acid receptors and results in increased levels of cGMP in target cells. Here we report that only a relatively short-term potentiation can be induced in the presence of nitro-L-arginine methyl ester (L-NAME), an NO synthase inhibitor. The effects of L-NAME on the duration of potentiation are partially reversed by coadministration of L-arginine, a precursor of neuronal NO, and by dibutyryl cGMP. Hemoglobin, which binds extracellular NO, also shortens the duration of stimulus-induced potentiation. The results suggest a role for NO in the maintenance of activity-dependent synaptic enhancements, possibly via the generation of cGMP.     

The effects of NMDA subunit composition on calcium influx and spike timing-dependent plasticity in striatal medium spiny neurons

Calcium through NMDA receptors (NMDARs) is necessary for the long-term potentiation (LTP) of synaptic strength; however, NMDARs differ in several properties that can influence the amount of calcium influx into the spine. These properties, such as sensitivity to magnesium block and conductance decay kinetics, change the receptor's response to spike timing dependent plasticity (STDP) protocols, and thereby shape synaptic integration and information processing. This study investigates the role of GluN2 subunit differences on spine calcium concentration during several STDP protocols in a model of a striatal medium spiny projection neuron (MSPN). The multi-compartment, multi-channel model exhibits firing frequency, spike width, and latency to first spike similar to current clamp data from mouse dorsal striatum MSPN. We find that NMDAR-mediated calcium is dependent on GluN2 subunit type, action potential timing, duration of somatic depolarization, and number of action potentials. Furthermore, the model demonstrates that in MSPNs, GluN2A and GluN2B control which STDP intervals allow for substantial calcium elevation in spines. The model predicts that blocking GluN2B subunits would modulate the range of intervals that cause long term potentiation. We confirmed this prediction experimentally, demonstrating that blocking GluN2B in the striatum, narrows the range of STDP intervals that cause long term potentiation. This ability of the GluN2 subunit to modulate the shape of the STDP curve could underlie the role that GluN2 subunits play in learning and development.     

Long-lasting increases in intrinsic excitability triggered by inhibition

Although experience-dependent changes in neural circuits are commonly assumed to be mediated by synaptic plasticity, modifications of intrinsic excitability may serve as a complementary mechanism. In whole-cell recordings from spontaneously firing vestibular nucleus neurons, brief periods of inhibitory synaptic stimulation or direct membrane hyperpolarization triggered long-lasting increases in spontaneous firing rates and firing responses to intracellular depolarization. These increases in excitability, termed firing rate potentiation, were induced by decreases in intracellular calcium and expressed as reductions in the sensitivity to the BK-type calcium-activated potassium channel blocker iberiotoxin. Firing rate potentiation is a novel form of cellular plasticity that could contribute to motor learning in the vestibulo-ocular reflex.     

Protein kinase C promotes N-methyl-D-aspartate (NMDA) receptor trafficking by indirectly triggering calcium/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation

Regulation of neuronal NMDA receptor (NMDAR) is critical in synaptic transmission and plasticity. Protein kinase C (PKC) promotes NMDAR trafficking to the cell surface via interaction with NMDAR-associated proteins (NAPs). Little is known, however, about the NAPs that are critical to PKC-induced NMDAR trafficking. Here, we showed that calcium/calmodulin-dependent protein kinase II (CaMKII) could be a NAP that mediates the potentiation of NMDAR trafficking by PKC. PKC activation promoted the level of autophosphorylated CaMKII and increased association with NMDARs, accompanied by functional NMDAR insertion, at postsynaptic sites. This potentiation, along with PKC-induced long term potentiation of the AMPA receptor-mediated response, was abolished by CaMKII antagonist or by disturbing the interaction between CaMKII and NR2A or NR2B. Further mutual occlusion experiments demonstrated that PKC and CaMKII share a common signaling pathway in the potentiation of NMDAR trafficking and long-term potentiation (LTP) induction. Our results revealed that PKC promotes NMDA receptor trafficking and induces synaptic plasticity through indirectly triggering CaMKII autophosphorylation and subsequent increased association with NMDARs.     

Short-term forms of presynaptic plasticity

Synapses exhibit several forms of short-term plasticity that play a multitude of computational roles. Short-term depression suppresses neurotransmitter release for hundreds of milliseconds to tens of seconds; facilitation and post-tetanic potentiation lead to synaptic enhancement lasting hundreds of milliseconds to minutes. Recent advances have provided insight into the mechanisms underlying these forms of plasticity. Vesicle depletion, as well as inactivation of both release sites and calcium channels, contribute to synaptic depression. Mechanisms of short-term enhancement include calcium channel facilitation, local depletion of calcium buffers, increases in the probability of release downstream of calcium influx, altered vesicle pool properties, and increases in quantal size. Moreover, there is a growing appreciation of the heterogeneity of vesicles and release sites and how they can contribute to use-dependent plasticity.     

Optical induction of synaptic plasticity using a light-sensitive channel

We have combined millisecond activation of channelrhodopsin-2 (ChR2), a light-gated ion channel, with two-photon calcium imaging to investigate active synaptic contacts in rat hippocampal slice cultures. Calcium influx was larger during light-induced action potentials than during action potentials induced by somatic current injection, leading to highly reproducible synaptic transmission. Pairing of light stimulation with postsynaptic depolarization induced long-term potentiation, making this technique ideal for genetic and pharmacological dissection of synaptic plasticity.     

Mechanisms for regulating synaptic efficiency in the visual cortex

Brief epochs of pairing of low frequency synaptic activation and postsynaptic depolarization, in vitro, in supragranular neurons of mature guinea-pig visual cortex lead to a transient (20–60 min) synaptic potentiation. This process is due to a true up-regulation of excitatory synapse efficiency onto the activated neuron. The potentiation requires NMDA receptor activation and a postsynaptic calcium signal for induction and it is modifiable by endogenous nitric oxide (NO) production in the mature cortex. In the cortex of young animals (< PND 21), the pairing-induced potentiation is robust and depends on a postsynaptic calcium signal but it is independent of NMDA receptor activation and NO production. The ability of cortical synaptosomes to release endogenous glutamate is enhanced by NMDA receptor activation and this enhancement is NO-dependent. The NO signal, however, does not amplify the glutamate release of all synapses but only those that have activated voltage-gated calcium channels and were presumably more active at the time of the NO signal. Electrophysiological recordings from visual cortical neurons in anesthetized cats with local iontophoresis of compounds that inhibit or facilitate endogenous cortical NO production reveal the capacity for NO to modulate visual responses in vivo. NO appears to act in the intact cortex by amplifying signals of visual inputs that were co-active at the time of the NO production. The adult visual cortex is capable of dramatic alterations in synaptic efficiency over brief periods suggesting a dynamic cortical network. NMDA receptors and nitric oxide contribute to these processes.     

Hidden prenatal malnutrition in the rat: role of β₁-adrenoceptors on synaptic plasticity in the frontal cortex

Moderate reduction in the protein content of the mother's diet (hidden malnutrition) does not alter body and brain weights of rat pups at birth, but leads to dysfunction of neocortical noradrenaline systems together with impaired long-term potentiation and visuo-spatial memory performance. As β?-adrenoceptors and downstream protein kinase signaling are critically involved in synaptic long-term potentiation and memory formation, we evaluated the β?-adrenoceptor density and the expression of cyclic-AMP dependent protein kinase, calcium/calmodulin-dependent protein kinase and protein kinase Fyn, in the frontal cortex of prenatally malnourished adult rats. In addition, we also studied if β?-adrenoceptor activation with the selective β? agonist dobutamine could improve deficits of prefrontal cortex long-term potentiation presenting these animals. Prenatally malnourished rats exhibited half of β?-adrenoceptor binding, together with a 51% and 65% reduction of cyclic AMP-dependent protein kinase α and calcium/calmodulin-dependent protein kinase α expression, respectively, as compared with eutrophic animals. Administration of the selective β? agonist dobutamine prior to tetanization completely rescued the ability of the prefrontal cortex to develop and maintain long-term potentiation in the malnourished rats. Results suggest that under-expression of neocortical β?-adrenoceptors and protein kinase signaling in hidden malnourished rats functionally affects the synaptic networks subserving prefrontal cortex long-term potentiation. β?-adrenoceptor activation was sufficient to fully recover neocortical plasticity in the PKA- and calcium/calmodulin-dependent protein kinase II-deficient undernourished rats, possibly by producing extra amounts of cAMP and/or by recruiting alternative signaling cascades.     

Recruitment of release sites underlies chemical presynaptic potentiation at hippocampal mossy fiber boutons

Synaptic plasticity is a cellular model for learning and memory. However, the expression mechanisms underlying presynaptic forms of plasticity are not well understood. Here, we investigate functional and structural correlates of presynaptic potentiation at large hippocampal mossy fiber boutons induced by the adenylyl cyclase activator forskolin. We performed 2-photon imaging of the genetically encoded glutamate sensor iGlu_u that revealed an increase in the surface area used for glutamate release at potentiated terminals. Time-gated stimulated emission depletion microscopy revealed no change in the coupling distance between P/Q-type calcium channels and release sites mapped by Munc13-1 cluster position. Finally, by high-pressure freezing and transmission electron microscopy analysis, we found a fast remodeling of synaptic ultrastructure at potentiated boutons: Synaptic vesicles dispersed in the terminal and accumulated at the active zones, while active zone density and synaptic complexity increased. We suggest that these rapid and early structural rearrangements might enable long-term increase in synaptic strength.

This study uses several high-resolution imaging techniques to investigate the structural correlates of presynaptic potentiation at hippocampal mossy fiber boutons, observing an increase in release sites and in release synchronicity accompanied by synaptic vesicle dispersion in the terminal and accumulation at release sites, but no modulation of the distance between calcium channel and release sites.     

Mitochondrial delivery is essential for synaptic potentiation

Mitochondria, as portable generators that power synaptic function, regulate the ATP supply and calcium homeostasis in the neuron. As molecular interactions within the synapses before and after the potentiation are beginning to be elucidated, the deciding moment during the tetanic stimulation that gives rise to the strengthening of the synapse remains a mystery. Here, I recorded electrically from an intact Drosophila nervous system, while simultaneously using time-lapse confocal microscopy to visualize mitochondria labeled with green fluorescent protein. I show that tetanic stimulation triggers a fast delivery of mitochondria to the synapse, which facilitates synaptic potentiation. Rotenone, an inhibitor of mitochondrial electron transport chain complex I, suppresses mitochondrial transport and abolishes the potentiation of the synapse. Expression of neurofibromin, which improves mitochondrial ATP synthesis in the neuron, enhances the movements of mitochondria to the synapse and promotes post-tetanic potentiation. These findings provide unprecedented evidence that the mitochondrial delivery to the synapse is critical for cellular learning.