Heterosynaptic interactions between septal and entorhinal inputs to the dentate gyrus: long-term potentiation effects

Cooperative interactions between neural pathways, in the production of long-term potentiation, may serve as models of associative memory. Brief, high-frequency activation of the septodentate input to dentate gyrus granule cells was found to produce only a short-term heterosynaptic potentiation of the perforant path-granule cell (PP-GC) population spike. Concurrent tetanization of perforant path and septodentate afferents, however, resulted in significantly greater long-term potentiation of the PP-GC population spike than was produced by tetanization of the perforant path alone.     

Long-trace interval eyeblink conditioning is impaired in mutant mice lacking the NMDA receptor subunit epsilon 1

To elucidate the role of the N-methyl-D-aspartate (NMDA) -type glutamate receptor subunit epsilon 1 (GluR epsilon 1) in classical eyeblink conditioning, delay and trace eyeblink conditioning were investigated in GluR epsilon 1-null mutant mice. In delay conditioning and short-trace interval conditioning with a trace interval of 250 ms, GluR epsilon 1 mutant mice attained a normal level of the conditioned response (CR), although acquisition was a little slower than in wild-type mice. In contrast, GluR epsilon 1 mutant mice exhibited severe impairment of the attained level of the CR and disturbed temporal pattern of CR expression in trace conditioning with a longer trace interval of 500 ms. These findings indicate that GluR epsilon 1 is essential for long-trace interval eyeblink conditioning. The impairments of the associative learning with a long temporal separation between the conditioned and unconditioned stimuli observed in the GluR epsilon 1 mutant mice could be attributed to an impairment of hippocampal long-term potentiation in this line of mutant mice.     

Impaired spatial working memory but spared spatial reference memory following functional loss of NMDA receptors in the dentate gyrus

Novel spatially restricted genetic manipulations can be used to assess contributions made by synaptic plasticity to learning and memory, not just selectively within the hippocampus, but even within specific hippocampal subfields. Here we generated genetically modified mice (NR1(deltaDG) mice) exhibiting complete loss of the NR1 subunit of the N-methyl-D-aspartate receptor specifically in the granule cells of the dentate gyrus. There was no evidence of any reduction in NR1 subunit levels in any of the other hippocampal subfields, or elsewhere in the brain. NR1(deltaDG) mice displayed severely impaired long-term potentiation (LTP) in both medial and lateral perforant path inputs to the dentate gyrus, whereas LTP was unchanged in CA3-to-CA1 cell synapses in hippocampal slices. Behavioural assessment of NR1(deltaDG) mice revealed a spatial working memory impairment on a three-from-six radial arm maze task despite normal hippocampus-dependent spatial reference memory acquisition and performance of the same task. This behavioural phenotype resembles that of NR1(deltaCA3) mice but differs from that of NR1(deltaCA1) mice which do show a spatial reference memory deficit, consistent with the idea of subfield-specific contributions to hippocampal information processing. Furthermore, this pattern of selective functional loss and sparing is the same as previously observed with the global GluR-A L-alpha-amino-3-hydroxy-5-methyl-4-isoxazelopropionate receptor subunit knockout, a mutation which blocks the expression of hippocampal LTP. The present results show that dissociations between spatial working memory and spatial reference memory can be induced by disrupting synaptic plasticity specifically and exclusively within the dentate gyrus subfield of the hippocampal formation.     

The effects of repeated induction of long-term potentiation in the dentate gyrus

Previous experiments have shown that long-term potentiation (LTP) generally lasts for only a few days or weeks. The LTP phenomenon would be more attractive as a memory model if it were more enduring. The experiments reported in this paper were designed to test the effects of repeated induction of LTP on the duration of LTP. Three groups of animals received 5 LTP-inducing stimulation sessions. In one group of animals, the stimulation sessions were administered every 24 h. In the 2nd group, the sessions were administered after the LTP effects had decayed to 50% of the peak values. In the 3rd group, the sessions were administered only after the response amplitudes had completely returned to pre-LTP baseline levels. None of the LTP measures were altered, in any group, as a result of repeated induction of LTP. The thresholds, the asymptotic levels of potentiation, and the decay rates were the same after each session. Other treatments may alter the duration of LTP effects, but repeated induction of LTP does not appear to have any lasting effects.     

Homosynaptic and heterosynaptic changes in driving of dentate gyrus interneurons after brief tetanic stimulation in vivo

This study addressed changes in interneuron driving in the dentate gyrus (DG) of urethane-anaesthetized rats in response to tetanic stimulation of the perforant path (PP) or the converging dentate commissural pathway (CP). Using an extracellular tungsten electrode, we recorded from putative interneurons in the DG that fired to stimulation of both the PP and the CP. Conditioning trains (400 Hz, 17.5 ms) were delivered to each pathway individually and to the two pathways together. The primary measure of synaptic drive was the latency of interneuron discharge. High-intensity PP tetany, CP tetany, and paired tetany consistently reduced firing latency to CP driving (P < .05 for all three), indicating an LTP-like increase in synaptic activation through the CP. High-intensity PP tetany decreased latency to PP driving in only two of seven cases. Heterosynaptic changes occurred frequently in individual experiments. Activity-mediated adjustments in synaptic driving of inhibitory interneurons could play a role in normal physiological function. © 1996 Wiley-Liss, Inc.     

Large-scale study of phosphoproteins involved in long-term potentiation in the rat dentate gyrus in vivo

The mechanisms underlying the induction of synaptic plasticity and the formation of long-term memory involve activation of cell-signalling cascades and protein modifications such as phosphorylation and dephosphorylation. Based on a protein candidate strategy, studies have identified several protein kinases and their substrates, which show an altered phosphorylation state during the early phases of long-term potentiation (LTP), yet only a limited number of synaptic phosphoproteins are known to be implicated in LTP. To identify new phosphoproteins associated with LTP, we have undertaken a proteomic study of phosphoproteins at different time points following the induction of LTP in the dentate gyrus in vivo (0, 15 and 90 min). For each time point, proteins from the dentate gyrus were separated by two-dimensional gel electrophoresis and stained with Pro−Q^® Diamond, a fluorescent stain specific for phosphoproteins. Fourteen proteins whose phosphorylation state varied significantly following LTP were identified using matrix-assisted laser desorption ionization/time of flight mass spectrometry and electrospray ionization-Orbitrap tandem mass spectrometry (MS/MS). They are involved in various cellular functions implicated in synaptic plasticity, such as intracellular signalling, axonal growth, exocytosis, protein synthesis and metabolism. Our results highlight new proteins whose phosphorylation or dephosphorylation is associated with LTP induction or maintenance. Further studies focusing on the regulation of specific phosphorylation sites will lead to greater understanding of the individual implications of these proteins in LTP as well as of their molecular interactions.     

The effects of kindling on GABA-mediated inhibition in the dentate gyrus of the rat. II. Receptor binding

Receptor binding estimates of the number of GABA and associated benzodiazepine (Bz) receptors were made in several brain regions following kindling. While the number of GABA receptors, as measured by both [³H]GABA and [³H]muscimol binding, was unaltered by kindling, Bz receptors were significantly increased in kindled amygdala and hippocampus. As the Bz receptor apparently functions to enhance GABA transmission, this finding suggests a possible mechanism for the kindling-induced enhancement of inhibition observed in previous evoked potential experiments.     

The supramammillary nucleus contributes to associative EPSP-spike potentiation in the rat dentate gyrus in vivo

The supramammillary nucleus (SUM) of the hypothalamus sends neural projections to the hippocampus and is supposed to be involved in learning and memory. To test the possibility that SUM afferents modulate hippocampal functions, we investigated the effect of electrical stimulation of the SUM on the induction of long-term potentiation (LTP) at medial perforant path (PP)--granule cell synapses in the dentate gyrus (DG) of anaesthetized rats. High-frequency stimulation of the SUM (100 pulses at 100 Hz) alone did not change PP--DG field potentials. However, when the SUM stimulation was applied simultaneously with weak tetanic stimulation of the PP (20 pulses at 20 Hz) which alone did not induce any potentiation, it produced a long-lasting potentiation of the population spike, without an accompanying increase in the population excitatory postsynaptic potential (EPSP). The EPSP-spike (E-S) potentiation induced by pairing SUM and PP stimulation was abolished by lesions of the fimbria--fornix, a major pathway of SUM afferents. SUM stimulation applied 1 s before or after PP stimulation failed to produce E-S potentiation, and SUM stimulation augmented PP--DG field potentials during tetanic stimulation. Furthermore, the E-S potentiation was abolished by blocking GABAergic neurotransmission with picrotoxin. These results suggest that coactivation of SUM and PP inputs produces a long-lasting increase of granule cell excitability by modulating GABAergic inhibition. SUM afferents may contribute to associative memory processing by modulating hippocampal excitability.     

Expression of the mu-opioid receptor is induced in dentate gyrus granule cells after focal cerebrocortical ischaemia and stimulation of entorhinal afferents

Focal ischaemia in the cerebral cortex affects the inducibility of long-term potentiation (LTP) in the hippocampus. This impairment of hippocampal function may result from excessive activation of cortico-hippocampal afferents and subsequent perturbation of hippocampal LTP-relevant transmitter systems, which include opioids. Here, we tested if permanent focal ischaemia and electrical afferent stimulation influence the expression of the mu-opioid receptor (MOR) in the rat hippocampus. In the applied ischaemia model, the entire ipsilateral cortical hemisphere and hippocampus experienced sustained excitation as indicated by a long-lasting increase in the expression of arg 3.1/arc (ARG) mRNA, a marker for neuronal activity. Expression of MOR mRNA and protein was strongly increased in granule cells, which contain very low MOR levels under normal conditions, but not in gamma-aminobutyric acid (GABA)ergic neurons, which express the MOR constitutively. In the molecular layer, which contains the dendrites of granule cells, focal ischaemia caused a redistribution of MOR-like immunoreactivity. In contrast to the dentate gyrus, MOR expression was unaltered in the hippocampus proper and in non-infarcted cortical areas. Repetitive high-frequency stimulation of cortico-hippocampal perforant path afferents induced strong MOR mRNA expression throughout the granular layer. However, weak tetanization sufficient to induce LTP and ARG expression did not influence MOR mRNA levels. Taken together, we provide direct evidence for the induction of MOR expression in granule cells experiencing sustained excitation by cortical afferents. In activated, MOR-expressing granule cells, inhibitory opioids may counter-regulate glutamatergic excitation by the perforant path.     

Long-term potentiation in the dentate gyrus: Effects of noradrenaline depletion in the awake rat

The chronic rat preparation was utilized to study the effects of noradrenaline (NA) depletion on field potentials recorded from the hilus of the fascia dentata. Both single pulses and high-frequency trains were applied to the perforant path (PP). The effects of NA depletion on baseline responses as well as on long-term potentiation (LTP) were examined. Reduced NA levels resulted in an increase in the population spike amplitude and a depression of the population excitatory postsynaptic potential (EPSP). Depleted animals showed significantly higher levels of LTP of the population EPSP, but reduced levels of population spike LTP (measured 13-15 min after tetanization). There were, however, no differences in LTP levels 1 week after the potentiation tests. These results demonstrate that NA levels do not affect that component of LTP which can persist for several weeks.     

Spike-timing-dependent plasticity at resting and conditioned lateral perforant path synapses on granule cells in the dentate gyrus: different roles of N-methyl-D-aspartate and group I metabotropic glutamate receptors

We examined the mechanisms underlying spike-timing-dependent plasticity induction at resting and conditioned lateral perforant pathway (LPP) synapses in the rat dentate gyrus. Two stimulating electrodes were placed in the outer third of the molecular layer and in the granule cell layer in hippocampal slices to evoke field excitatory postsynaptic potentials (fEPSPs) and antidromic field somatic spikes (afSSs), respectively. Long-term potentiation (LTP) of LPP synapses was induced by paired stimulation with fEPSP preceding afSS. Reversal of the temporal order of fEPSP and afSS stimulation resulted in long-term depression (LTD). Induction of LTP or LTD was blocked by D,L-2-amino-5-phosphonopentanoic acid (AP5), showing that both effects were N-methyl-D-aspartate receptor (NMDAR)-dependent. Induction of LTP was also blocked by inhibitors of calcium-calmodulin kinase II, protein kinase C or mitogen-activated/extracellular-signal regulated kinase, suggesting that these are downstream effectors of NMDAR activation, whereas induction of LTD was blocked by inhibitors of protein kinase C and protein phosphatase 2B. At LPP synapses previously potentiated by high-frequency stimulation or depressed by low-frequency stimulation, paired fEPSP-afSS stimulation resulted in 'de-depression' at depressed LPP synapses but had no effect on potentiated synapses, whereas reversal of the temporal order of fEPSP-afSS stimulation resulted in 'de-potentiation' at potentiated synapses but had no effect on depressed synapses. Induction of de-depression and de-potentiation was unaffected by ap5 but was blocked by 2-methyl-6-(phenylethynyl) pyridine hydrochloride, a group I metabotropic glutamate receptor blocker, showing that both were NMDAR-independent but group I metabotropic glutamate receptor-dependent. In conclusion, our results show that spike-timing-dependent plasticity can occur at both resting and conditioned LPP synapses, its induction in the former case being NMDAR-dependent and, in the latter, group I metabotropic glutamate receptor-dependent.     

Different chronic cocaine administration protocols induce changes on dentate gyrus plasticity and hippocampal dependent behavior

Hippocampus is a limbic structure that participates in learning and memory formation. Specifically the dentate gyrus has been described as a hippocampal subregion with high rates of plasticity and it is targeted by different psychoactive drugs modulating synaptic plasticity. Repeated cocaine administration induces sensitization to the locomotor effects and it is believed that sensitization involves the same mechanisms of drug seeking and relapse. Although, the mechanisms underlying sensitization is not fully understood. In this work we investigated the impact of repeated intraperitoneal administration of cocaine (15 or 20 mg/kg/day along 5 or 15 days respectively; and 15 mg/kg/day along 5 day followed by a challenge dose after three days of withdrawal) on the dentate gyrus synaptic plasticity, differentiating between sensitized and nonsensitized rats. Furthermore, we correlated changes on the hippocampal synaptic plasticity to memory retention. Our results revealed that the prevalence of cocaine sensitization (around 50%) was identical in all protocols used. The results found in the threshold to generate LTP were similar for all protocols used, being the threshold values cocaine‐treated groups (sensitized and nonsensitized) significantly reduced compared to controls, observing the highest reduction in the sensitized group. Moreover, we observed a facilitated retention of recent memory formation only in sensitized animals the nonsensitized subjects remained at the control levels. In conclusion, sensitization to cocaine generates a high efficiency of hippocampal synaptic plasticity that may underlie the aberrant engagement of learning processes occurred during drug addiction. Synapse 64:742–753, 2010. © 2010 Wiley‐Liss, Inc.     

Time-dependent induction of depotentiation in the dentate gyrus of freely moving rats: involvement of group 2 metabotropic glutamate receptors

Depotentiation comprises a reversal of tetanization-induced long-term potentiation (LTP) which occurs following low-frequency stimulation (LFS) in the hippocampus in vivo. Although depotentiation has been consistently demonstrated in the CA1 region, no positive reports of the existence of depotentiation in the dentate gyrus in vivo have occurred. This study therefore investigated whether depotentiation is possible in the dentate gyrus in vivo. We found that depotentiation can be induced, but it is very tightly dependent on the interval between tetanization and LFS. Thus, LFS given 2 or 5 min following tetanization produced significant depotentiation, whereas LFS given 10-30 min following tetanization had no significant effect on the expression of LTP. Depotentiation occurred in two phases: a transient depression of evoked responses to below pre-tetanization values, which occurred in the first 60 min following LFS, and a recovery of this response to a stable level of synaptic transmission which comprised a significant reduction in the magnitude of LTP. Group 2 metabotropic glutamate receptors (mGluRs) play an important role in the expression of long-term depression in vivo. We therefore investigated whether group 2 mGluRs contribute to depotentiation. The group 2 antagonist (2S)-alpha-ethylglutamic acid (EGLU) inhibited the early transient depression at a concentration which inhibits LTD in vivo, but did not block the expression of depotentiation. EGLU also inhibited the transient depression induced by 5 Hz given alone. Increasing the concentration of EGLU prevented depotentiation, however. The group 2 agonist (S)-4-carboxy-3-hydroxyphenyl- glycine (4C3HPG) inhibited LTP and enhanced depotentiation. These data suggest a role for group 2 mGluRs in depotentiation.     

Enhanced long-term potentiation induced in rat dentate gyrus by coactivation of septal and entorhinal inputs: Temporal constraints

High-frequency activation of the entorhinal cortical (perforant path) inputs to the rat dentate gyrus can produce a long-term potentiation (LTP) of perforant path-dentate evoked responses. In this paper we examined the enhanced LTP effects produced by coactivation of septal and entorhinal inputs to the dentate gyrus. Trains of electrical stimulation applied to the two inputs were found to increase the magnitude of LTP to a level above that produced by trains applied to the perforant path alone. The largest LTP increments were observed when the septal trains were applied less than 100 ms prior to the perforant path trains. If the septal trains followed the perforant path trains there was no additional increment in LTP magnitude, regardless of the intertrain interval. The relationship of this cooperativity effect to mechanisms of associative learning is discussed.     

Blockade of norepinephrine-induced long-lasting potentiation in the hippocampal dentate gyrus by an inhibitor of protein synthesis

The mechanism of action of norepinephrine (NE)-induced potentiation of the population spike in the dentate gyrus of hippocampal slices was examined and compared with NE effects in field CA1. NE-induced potentiation was confined to the dentate gyrus, where slices perfused for 30 min with concentrations of NE as low as 5 microM exhibited potentiation of the perforant path evoked population spike. Potentiation began within 15 min, and lasted many hours after NE was washed out. Experiments where slices were pre-incubated with the protein synthesis inhibitor emetine indicated that there are two distinct phases to NE-induced potentiation. The initial short-term NE-induced potentiation (NEP) seen during NE application was not affected by a 30 min pre-incubation with emetine, whereas the long-lasting potentiation (NELLP) which persists after NE washout was completely blocked by emetine at a concentration which we have previously shown to be effective in blocking hippocampal long-term potentiation (LTP). Additional experiments indicated that both phases of NE-induced potentiation were completely blocked by the beta-antagonist propranolol and the beta 1-antagonist metoprolol. Furthermore, pre-incubation of slices with the direct-acting adenylate cyclase stimulant forskolin shifted the dose-response curves for both phases of NE-induced potentiation to the left. These results suggest that NE-induced potentiation is probably mediated by beta 1-receptor stimulation of adenylate cyclase. We have previously shown an importance for beta 1-receptor stimulation of adenylate cyclase in the production of LTP in the dentate. Thus, these results demonstrate a number of similarities between hippocampal LTP and NELLP in the dentate gyrus.     

Field potential recordings in dentate gyrus of anesthetized rats: Stability of baseline

Urethane is a standard anesthetic utilized for in vivo recordings in the hippocampus. In studies of long‐term potentiation (LTP), the measure of interest is the response amplitude minutes to hours following train delivery. In the absence of experimental treatment, we have consistently observed upward drift in the amplitude of the population spike (PS) and EPSP slope of the dentate gyrus (DG) evoked field response in acute surgical preparations performed in the urethanized rat. The present study systematically monitored PS amplitude and EPSP slope in the DG every 30 minutes for 6 hours following optimal positioning of Teflon‐coated bipolar stainless steel electrodes under urethane anesthesia. At maximal stimulus intensities, large time‐dependent increases in PS amplitude (70–80%) were observed over the first 2–4 hours, an effect that was exaggerated at lower stimulus intensities. Increases in the EPSP slope were smaller in magnitude (20–30%) and stabilized within a shorter period of time (1–2 hours). Animals were warmed on a heating pad and body and brain temperature remained constant over the recording session. Reducing stimulating electrode size and recording with glass micropipettes did not alleviate the upward drift in response amplitude. Similar increases were also seen under pentobarbital anesthesia. To dissociate anesthetic from surgical effects, recordings were obtained from animals previously prepared with indwelling electrodes and injected with urethane. Although slight declines (10–15%) in EPSP slope occurred over time, no significant alterations in PS amplitude were seen in the chronic preparation at high stimulus intensities. Low stimulus intensities yielded a more variable response pattern and, in direct contrast to the acute preparation, time‐dependent declines, not increases, were noted in both parameters. These data suggest that generalized surgical trauma contributes to the upward drift in response amplitude and indicate that long stabilization periods are required in acute surgical preparations for accurate field potential recordings. Hippocampus 1999;9:277–287. Published 1999 Wiley‐Liss, Inc.     

Pathway specificity of noradrenergic plasticity in the dentate gyrus

Previous experiments have described highly specific effects of noradrenergic agonists on synaptic transmission in the dentate gyrus (DG). For example, perfusion of hippocampal slices with the beta-noradrenergic agonist isoproterenol induces a long-lasting potentiation (LLP) of extracellularly recorded responses following stimulation of the medial perforant path (PP), and long-lasting depression (LLD) of responses evoked by stimulation of the lateral PP (Dahl D, Sarvey JM, 1989, Proc Natl Acad Sci USA 86:4776–4780). To examine the possible interactions of LLP, LLD, and long-term potentiation induced by tetanic stimulation (LTP), the authors recorded extracellular field potentials evoked in the DG by stimulation of the lateral or medial perforant path following LTP and LLP or LLD, invoked in different orders. After establishment of LLP or LLD by path application of isoproternol, subsequent tetanization of the respective afferents resulted in additional potentiation of the medial PP-evoked response and return of the lateral PP-evoked response to baseline levels. In other slices application of isoproterenol after establishment of LTP resulted in further potentiation of medial PP-evoked responses but no change in the potentiated response evoked by lateral PP stimulation. Thus the pathway specificity was maintained irrespective of the history of previous potentiation or depression. Experiments using the specific beta₁ antagonist metoprolol further confirmed pathway specificity. Perfusion with 20 μM of metoprolol appeared to reduce LTP evoked by stimulation of the medial but not lateral PP. In a subsequent experiment, metoprolol in the absence of tetanization produced LLD of the medial PP-evoked response and LLP of the lateral PP-evoked response, opposite to the effects of ISO. These results confirm the impressive extent of pathway specificity in the DG and reveal the persistent capacity for synaptic modification as exemplified by the processes of LLP, LLD, and LTP. © 1994 Wiley-Liss, Inc.     

Long-term potentiation in the rat dentate gyrus is associated with enhanced Arc/Arg3.1 protein expression in spines, dendrites and glia

Electron microscopic immunocytochemical methods were used to determine the localization, subcellular distribution and expression of activity-regulated cytoskeletal protein (Arc/Arg3.1) in dentate gyrus after unilateral induction of long-term potentiation (LTP) in the perforant pathway of anaesthetized rats. At 2 h post-induction, immunoreaction product was visible in the dentate gyrus in both the granule cell and molecular layers. Arc expression was higher in the potentiated than the unstimulated contralateral hemisphere. Single-section electron microscopy analysis in unstimulated tissue and in tissue prepared 2 and 4 h after LTP induction showed Arc immunoreactivity (Arc-IR) in dendrites, dendritic spines and glia. Arc-IR was associated with synaptic and non-synaptic plasma membrane apposed to axon terminals and with cytoplasmic organelles, including the cytoskeleton. Arc-IR was also present in neuronal perikarya and there was occasional labelling of nuclei and axons. At 2 h post-LTP induction, there were significant increases in Arc-IR within the granule cell and molecular layers of the dentate gyrus and particularly within the middle molecular layer relative to the inner and outer molecular layers. This increase in Arc expression 2 h after LTP induction was blocked by the N-methyl-D-aspartate receptor antagonist (RS)-3-2-carboxypiperazin-4-yl-propyl-1-phosphonic acid. In animals killed 4 h after LTP induction, Arc expression had declined and differences between the potentiated and unpotentiated hemispheres were no longer significant. Our data provide ultrastructural evidence for a transient LTP-associated increase in the expression of Arc protein in the middle molecular layer of the dentate gyrus, with preferential targeting to dendrites, dendritic spines and glia.