Enhancement of long-term depression by soluble amyloid ß protein in rat hippocampus is mediated by metabotropic glutamate receptor and involves activation of p38MAPK, STEP and caspase-3.

It is reported that the amyloid-ß protein (Aß)-induced impairments in synaptic plasticity coincide with memory decline and dementia. Although Aß-induced inhibition of hippocampal long-term potentiation has been intensively investigated, the underlying mechanism of Aß-enhanced long-term depression (LTD) is not clear. Here, we report that acute exposure of rat hippocampal slices to soluble Aß-enhanced LTD induced by weak low-frequency stimulation (wLFS; 1Hz for 3 min, 180 pulses) in granule cells of the dentate gyrus. Application of LY341495 (a non-selective Group I/II metrabotropic glumate receptor (mGluR) antagonist) completely blocked Aß-enhanced LTD, whereas D-AP5 (a not selective N-methyl-d-aspartate receptor (NMDAR) antagonist) had no effect on Aß-enhanced LTD compared with controls. In addition, Aß-enhanced LTD was occluded by pre-application of 3,5-dihydroxyphenylglycine, a Group1 mGluR (mGluR1/5) agonist, suggesting Aß-enhanced LTD depends on mGluR1/5 but not NMDAR. We also report here that p38 mitogen-activated protein kinase (p38MAPK) inhibitor SB203580 and postsynaptic protein tyrosine phosphatase inhibitors phenylarsine oxide and sodium orthovanadate prevented the facilitatory effect of Aß on LTD. Application of striatal-enriched protein tyrosine phosphatase (STEP) activator MG132 facilitated induction of LTD by wLFS, but did not block following Aß-enhanced LTD induced by another wLFS. On the other hand, Aß-enhanced LTD blocked following MG132-LTD by wLFS, suggesting Aß-enhanced hippocampal LTD involves STEP activation. Application of either non-selective caspase inhibitor Z-VAD-FMK or caspase-3 selective inhibitor Z-DEVD-FMK prevented Aß-enhanced LTD. However, neither the tumor necrosis factor-α converting enzyme inhibitor TAPI-2 nor the mammalian target of rapamycin inhibitor rapamycin prevented the enhancement of Aß on LTD. Therefore, we conclude that soluble Aß enhances LTD in the hippocampal dentate gyrus region, and the facilitatory effect of Aß on LTD involves mGluR1/5, p38MAPK, STEP and caspase-3 activation.

Metabotropic glutamate receptor-dependent long-term potentiation.

The induction of the most common form of LTP is well known to involve activation of N-methyl-D-aspartate receptors. However, considerable evidence has also shown that certain forms of LTP induction at excitatory synapses onto both principle cells and interneurons are dependent on activation of metabotropic glutamate receptors (mGluRs). mGluR-dependent LTP occurs in widespread areas of the brain including the neocortex, hippocampus, striatum and nucleus accumbens. mGluR-dependent forms of LTP have been found to be diverse, involving activation of mGluR1 or mGluR5 and can be of AMPAR-mediated transmission or of NMDAR-mediated transmission. Furthermore, the mGluR-dependent LTP may involve activation of other receptors, in particular, activation of NMDAR, dopamine and adenosine receptors. mGluR-dependent LTP can be expressed presynaptically or postsynaptically, and can involve a range of intracellular mediators including protein kinase C (PKC) and protein kinase A (PKA), tyrosine kinase Src and nitric oxide (NO).

A role for protein kinase A and protein kinase M zeta in muscarinic acetylcholine receptor-initiated persistent synaptic enhancement in rat hippocampus in vivo.

Antagonists at presynaptic muscarinic autoreceptors increase endogenous acetylcholine (ACh) release and enhance cognition but little is known regarding their actions on plasticity at glutamatergic synapses. Here the mechanisms of the persistent enhancement of hippocampal excitatory transmission induced by the M2/M4 muscarinic ACh receptor antagonist methoctramine were investigated in vivo. The persistent facilitatory effect of i.c.v. methoctramine in the CA1 region of urethane-anesthetized rats was mimicked by gallamine, an M2 receptor antagonist, supporting a role for this receptor subtype. Neither the N-methyl-D-aspartate (NMDA) receptor antagonists D-(-)-2-amino phosphonopentanoic acid (d-AP5) and memantine, nor the metabotropic glutamate receptor subtype 1a antagonist (S)-(+)-alpha-amino-4-carboxy-2-methylbenzeneacetic acid (LY367385) significantly affected the methoctramine-induced persistent synaptic enhancement, indicating a lack of requirement for these glutamate receptors. The selective kinase inhibitors Rp-adenosine-3', 5'-cyclic monophosphorothioate (Rp-cAMPS) and the myrostylated pseudosubstrate peptide, Myr-Ser-Ile-Tyr-Arg-Arg-Gly-Ala-Arg-Arg-Trp-Arg-Lys-Leu-OH (ZIP), were used to investigate the roles of protein kinase A (PKA) and the atypical protein kinase C, protein kinase Mzeta (PKM zeta), respectively. Remarkably, pretreatment with either agent prevented the induction of the persistent synaptic enhancement by methoctramine and post-methoctramine treatment with Rp-cAMPS transiently reversed the enhancement. These findings are strong evidence that antagonism of M2 muscarinic ACh receptors in vivo induces an NMDA receptor-independent persistent synaptic enhancement that requires activation of both PKA and PKM zeta.

Synaptic memory mechanisms: Alzheimer's disease amyloid beta-peptide-induced dysfunction.

There is growing evidence that mild cognitive impairment in early AD (Alzheimer's disease) may be due to synaptic dysfunction caused by the accumulation of non-fibrillar, oligomeric Abeta (amyloid beta-peptide), long before widespread synaptic loss and neurodegeneration occurs. Soluble Abeta oligomers can rapidly disrupt synaptic memory mechanisms at extremely low concentrations via stress-activated kinases and oxidative/nitrosative stress mediators. Here, we summarize experiments that investigated whether certain putative receptors for Abeta, the alphav integrin extracellular cell matrix-binding protein and the cytokine TNFalpha (tumour necrosis factor alpha) type-1 death receptor mediate Abeta oligomer-induced inhibition of LTP (long-term potentiation). Ligands that neutralize TNFalpha or genetic knockout of TNF-R1s (type-1 TNFalpha receptors) prevented Abeta-triggered inhibition of LTP in hippocampal slices. Similarly, antibodies to alphav-containing integrins abrogated LTP block by Abeta. Protection against the synaptic plasticity-disruptive effects of soluble Abeta was also achieved using systemically administered small molecules targeting these mechanisms in vivo. Taken together, this research lends support to therapeutic trials of drugs antagonizing synaptic plasticity-disrupting actions of Abeta oligomers in preclinical AD.

Muscarinic acetylcholine receptor-dependent induction of persistent synaptic enhancement in rat hippocampus in vivo.

Presynaptic terminal autoinhibitory muscarinic acetylcholine (ACh) receptors are predominantly of the M2/M4 subtypes and antagonists at these receptors may facilitate cognitive processes by increasing ACh release. The present study examined the ability of the M2/M4 muscarinic ACh receptor antagonist N,N'-bis [6-[[(2-methoxyphenyl)methyl]amino]hexyl]-1,8-octane diamine tetrahydrochloride (methoctramine) to induce and modulate synaptic plasticity in the CA1 area of the hippocampus in urethane-anesthetized rats. Both methoctramine and another M2/M4 antagonist, {11-[[2-[(diethylamino)methyl]-1-piperidinyl]acetyl]-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepin-6-one} (AF-DX 116), caused a rapid onset and persistent increase in baseline synaptic transmission after i.c.v. injection. Consistent with a requirement for activation of non-M2 receptors by endogenously released ACh, the M1/M3 receptor selective antagonists 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) and 4,9-dihydro-3-methyl-4-[(4-methyl-1-piperazinyl)acetyl]-10H-thieno[3,4-b][1,5]benzodiazepin-10-one dihydrochloride (telenzepine) prevented the induction of the persistent synaptic enhancement by methoctramine. The requirement for cholinergic activation was transient and independent of nicotinic ACh receptor stimulation. The synaptic enhancement was inhibited by the prior induction of long-term potentiation (LTP) by high frequency stimulation but induction of the synaptic enhancement by methoctramine before high frequency stimulation did not inhibit LTP. Unlike high frequency stimulation-evoked LTP, the synaptic enhancement induced by methoctramine appeared to be NMDA receptor-independent. The present studies provide evidence for the rapid induction of a persistent potentiation at hippocampal glutamatergic synapses by endogenous ACh in vivo following disinhibition of inhibitory M2 muscarinic autoreceptors.

Developmental dependence, the role of the kinases p38 MAPK and PKC, and the involvement of tumor necrosis factor-R1 in the induction of mGlu-5 LTD in the dentate gyrus.

The mechanisms of mGluR-LTD were studied in the dentate gyrus in vitro. The most effective protocol for inducing mGluR-LTD in 6-8 week animals was brief high frequency stimulation (HFS) applied in the presence of the NMDAR antagonist AP5. Evidence for HFS inducing LTD via activation of perisynaptically located mGluRs was established, as an inhibitor of glutamate transporter potentiated HFS-LTD. HFS-LTD was mainly mediated by activation of mGluR5, although a partial involvement of mGluR1 was found. (RS)-3,5-Dihydroxyphenylglycine (DHPG) also induced LTD, but in an age dependent manner, being large in 2 week animals but absent in 6-8 week animals. DHPG-LTD in the dentate gyrus also had a much slower rise time than that in CA1, and unlike CA1, the expression/maintenance of mGluR-LTD was not inhibited by mGluR antagonists. The use of pharmacological inhibitors showed that the induction of HFS-LTD was partially dependent upon activation of L-type Ca channels, release of Ca from ryanodine receptor-sensitive intracellular Ca stores, and the kinases p38 mitogen-activated protein kinase (MAPK), protein kinase C (PKC), but not c-Jun N-terminal kinase or COX-2. Evidence for the involvement of tumor necrosis factor-receptor 1 (TNF-R1) in the induction of mGluR-LTD was presented in the present study, with both HFS-mGluR-LTD and DHPG-LTD being absent in mutant mice null for TNF-R1.

Synaptic plasticity disruption by amyloid beta protein: modulation by potential Alzheimer's disease modifying therapies.

AD (Alzheimer's disease) is characterized by a progressive and devastating mental decline that is usually presaged by impairment of a form of memory dependent on medial temporal lobe structures, including the hippocampus. The severity of clinical dementia correlates positively with the cerebral load of the AD-related protein Abeta (amyloid beta), particularly in its soluble form rather than the insoluble fibrillar Abeta found in amyloid plaques. Recent research in animal models of AD has pointed to a potentially important role for rapid disruptive effects of soluble species of Abeta on neural function in causing a relatively selective impairment of memory early in the disease. Our experiments assessing the mechanisms of Abeta inhibition of LTP (long-term potentiation), a correlate of memory-related synaptic plasticity, in the rodent hippocampus showed that low-n oligomers were the soluble Abeta species primarily responsible for the disruption of synaptic plasticity in vivo. Exogenously applied and endogenously generated anti-Abeta antibodies rapidly neutralized and prevented the synaptic plasticity disrupting effects of these very potent Abeta oligomers. This suggests that active or passive immunotherapeutic strategies for early AD should target Abeta oligomers in the brain. The ability of agents that reduce nitrosative/oxidative stress or antagonize stress-activated kinases to prevent Abeta inhibition of LTP in vitro points to a key role of these cellular mechanisms at very early stages in Abeta-induced neuronal dysfunction. A combination of antibody-mediated inactivation of Abeta oligomers and pharmacological prevention of cellular stress mechanisms underlying their synaptic plasticity disrupting effects provides an attractive strategy in the prevention of early AD.

Synaptically stimulated induction of group I metabotropic glutamate receptor-dependent long-term depression and depotentiation is inhibited by prior activation of metabotropic glutamate receptors and protein kinase C.

We have investigated metaplasticity of the group I metabotropic glutamate receptor (mGluR)-dependent long-term depression (LTD) and depotentiation (DP) induced by physiological synaptic stimulation in the medial perforant path of the dentate gyrus in vitro. Group I mGluR-LTD/DP was inhibited by prior preconditioning brief high frequency stimulation (HFS) if the preconditioning HFS induced long-term potentiation (LTP) or if the induction of LTP was inhibited by an NMDA receptor antagonist. The inhibitory effect of the preconditioning HFS on LTD/DP was dependent upon activation of mGluRs, as it was blocked by the presence of the mGluR antagonist (S)-alpha-methyl-4-carboxyphenylglycine during the preconditioning stimulation. The inhibitory effect of the preconditioning HFS involved stimulation of PKC, as the presence of the PKC inhibitor bisindolylmaleimide (BIS) during the preconditioning stimulation prevented the inhibitory effect of such preconditioning stimulation. Activation of PKC was also necessary for the induction of mGluR-LTD itself, as the PKC inhibitor BIS prevented the induction of the mGluR-LTD. We suggest that the physiological stimulation of mGluRs by the preconditioning stimulation produces a PKC-dependent inactivation of subsequent group I mGluR functioning and thereby an inhibition of induction of group I mGluR-dependent LTD/DP induction.

Mechanisms underlying the inhibition of long-term potentiation by preconditioning stimulation in the hippocampus in vitro.

We have investigated the mechanisms underlying a form of metaplasticity, namely the inhibition by preconditioning stimulation of high frequency stimulation (HFS)-induced long-term potentiation (LTP) in the medial perforant path of the dentate gyrus. Preconditioning stimulation (weak 50 Hz) was found to inhibit subsequent LTP induction if applied 10-20 min, but not 2 or 45 min, prior to the HFS. Preconditioning stimulation in the form of low frequency stimulation did not block LTP induction. The inhibition of LTP was not caused by a reduction in N-methyl-D-aspartate receptor (NMDAR) transmission, as the preconditioning stimulation did not reduce isolated NMDAR-mediated EPSPs. The involvement of group I and group II metabotropic glutamate receptor (mGluR) activation in the inhibition of LTP was demonstrated by experiments in which the inhibition of LTP by the preconditioning stimulation was prevented by the presence of antagonists of group I or group II mGluR during the preconditioning stimulation. Moreover, group I and group II mGluR agonists directly inhibited subsequent LTP induction. The involvement of NMDAR in the preconditioning stimulation was shown by the ability of an NMDAR antagonist to prevent the inhibition of LTP by the preconditioning stimulation. The preconditioning inhibition of LTP induction was shown by the use of kinase inhibitors to involve activation of PKC and p38 MAP kinase, but not p42 MAP kinase or tyrosine kinase. We conclude that the preconditioning inhibition of LTP induction is a complex process which involves activation of NMDAR, group I and group II mGluR, and intracellular cascades activating PKC and p38 MAP kinase.

Early spatial memory deficit induced by 2,5-hexanedione in the rat.

2,5-Hexanedione (2,5-HD), the major common neurotoxic metabolite of n-hexane and methyl n-butyl ketone, causes a delayed neuropathy with associated sensorimotor impairments. The question arises as to whether specific cognitive deficits occur even prior to changes in sensorimotor ability. The present experiments examined the effects of 2,5-HD on spatial navigation of rats in a water maze at levels/times that did not affect spontaneous exploratory motor activity in an open field holeboard apparatus. Exposure to 1% 2,5-HD in the drinking water for 2 weeks did not significantly affect escape learning, as measured by latency to find a hidden platform. However, 2,5-HD treated animals were impaired in the use of a spatial strategy during a recall test. A similar impairment in spatial memory was observed after i.p. injection of 500 mg/kg/day 2,5-HD for 4 days, in the absence of significant changes in sensorimotor ability or weight loss. Thus 2,5-HD may mediate some of the cognitive effects of hexacarbons and these changes can occur prior to the development of motor symptoms.

Activation of group II metabotropic glutamate receptors results in long-term potentiation following preconditioning stimulation in the dentate gyrus.

The role of group II metabotropic glutamate receptors in the induction/expression of long-term potentiation has been investigated in the medial perforant path of the outer (infrapyramidal) blade of the rat dentate gyrus in vitro. Activation of group II metabotropic glutamate receptors by perfusion of the selective agonist LY354740 did not induce long-term potentiation or long-term depression in control. However, LY354740, applied following the induction of long-term potentiation by high frequency stimulation, resulted in additional long-term potentiation. LY354740 was only found to cause additional long-term potentiation if the pre-existing high frequency stimulation-induced long-term potentiation was sub-maximal. Although activation of metabotropic glutamate receptors was not required for induction of high frequency stimulation-induced long-term potentiation, activation of both group I and group II metabotropic glutamate receptors was required during high frequency stimulation-induced long-term potentiation in order for subsequent application of LY354740 to result in additional long-term potentiation. Thus, the long-term potentiation caused by application of LY354740 following high frequency-induced long-term potentiation was prevented if the high frequency stimulation was given in the presence of (S)-alpha-methyl-4-carboxyphenylglycine or the selective group I or group II metabotropic glutamate receptor antagonists 1-aminoindan-1,5-dicarboxylic acid or (2S)-alpha-ethylglutamic acid respectively. The long-term potentiation caused by LY354740 was also dependent upon activation of N-methyl-D-aspartate receptors during the high frequency stimulation, being blocked if high frequency stimulation was given in the presence of the N-methyl-D-aspartate receptor antagonist, D(-)-2-amino-5-phosphonopentanoic acid. The long-term potentiation resulting from activation of group II metabotropic glutamate receptors could be due either to the enhancement of the expression level of the high frequency stimulation-induced long-term potentiation, or alternatively, to a direct novel induction of long-term potentiation. In either theory, the long-term potentiation resulting from activation of group II metabotropic glutamate receptors is dependent upon prestimulation of group I and group II metabotropic glutamate receptors and N-methyl-D-aspartate receptors during the 'preconditioning high frequency stimulation'.

Presynaptic group II mGluR inhibition of short-term depression in the medial perforant path of the dentate gyrus in vitro.

Inhibition of short-term plasticity by activation of presynaptic group II metabotropic glutamate receptors (group II mGluR) was investigated in the medial perforant path of the dentate gyrus in the hippocampus in vitro. Brief trains of stimulation (10 stimuli at 1--200 Hz) evoked short-term depression of field excitatory postsynaptic potentials (EPSPs). The steady-state level of depression, measured after 10 stimuli, was frequency dependent, increasing between 1 and 200 Hz. Activation of group II mGluR by the selective agonist LY354740 did not alter short-term depression evoked by frequencies up to 10 Hz, but did inhibit short-term depression evoked at higher frequencies in a frequency- and concentration-dependent manner. The time-averaged postsynaptic response (EPSP per unit time) was found to increase linearly with frequency up to approximately 20 Hz. At higher frequencies, the response plateaued, thereby becoming independent of frequency. Frequencies above this were differentiated only during the transient postsynaptic response that accompanies changes in firing rates. Activation of presynaptically located group II mGluR increased the frequency at which the EPSP per unit time plateaued up to 30-50 Hz.

NMDA receptor- and metabotropic glutamate receptor-dependent synaptic plasticity induced by high frequency stimulation in the rat dentate gyrus in vitro.

1. The mechanisms of long-term potentiation (LTP) and long-term depression (LTD) induced by brief high frequency stimulation (HFS), paired with a particular pattern and amplitude of depolarisation has been investigated in the medial perforant pathway of the dentate gyrus of the 2- to 3-week-old rat hippocampus in vitro. 2. N-Methyl-D-aspartate (NMDA) receptor (NMDAR) activation was measured quantitatively during HFS-induced NMDAR-dependent LTP, LTD and at the LTD--LTP crossover point in order to test the hypothesis that the induction of the particular form of plasticity depends on the intensity of NMDAR activation. 3. The induction of LTD, the LTD--LTP crossover point and LTP was associated with an increasing NMDAR charge transfer. 4. In addition to the NMDAR-dependent LTD, a group I metabotropic glutamate receptor (mGluR)-dependent LTD could be induced by high intensity HFS paired with depolarisation under conditions of NMDAR inhibition. 5. The induction of mGluR-dependent LTD requires membrane depolarisation, Ca(2+) influx via L-type Ca(2+) channels and a rise in intracellular Ca(2+). 6. Quantal analysis involving minimal stimulation demonstrated that the mGluR-dependent LTD induction was associated with a decrease in potency and an increase in failure rate.

Application of N-methyl-D-aspartate induces long-term potentiation in the medial perforant path and long-term depression in the lateral perforant path of the rat dentate gyrus in vitro.

The effect of application of N-methyl-D-aspartate (NMDA) on synaptic plasticity was studied in the medial and lateral perforant path-granule cell synapse in the outer blade of the dentate gyrus in vitro. Field excitatory post-synaptic potentials were recorded from the middle or outer molecular layer in response to stimulation of the medial or lateral perforant path. Bath perfusion of NMDA (10 microM, 5 min) resulted in induction of long-term potentiation in the medial perforant path, and induction of long-term depression in the lateral perforant path.

Use-dependent effects of amyloidogenic fragments of (beta)-amyloid precursor protein on synaptic plasticity in rat hippocampus in vivo.

The Alzheimer's disease-related beta-amyloid precursor protein (beta-APP) is metabolized to a number of potentially amyloidogenic peptides that are believed to be pathogenic. Application of relatively low concentrations of the soluble forms of these peptides has previously been shown to block high-frequency stimulation-induced long-term potentiation (LTP) of glutamatergic transmission in the hippocampus. The present experiments examined how these peptides affect low-frequency stimulation-induced long-term depression (LTD) and the reversal of LTP (depotentiation). We discovered that beta-amyloid peptide (Abeta1-42) and the Abeta-containing C -terminus of beta-APP (CT) facilitate the induction of LTD in the CA1 area of the intact rat hippocampus. The LTD was frequency- and NMDA receptor-dependent. Thus, although low-frequency stimulation alone was ineffective, after intracerebroventricular injection of Abeta1-42, it induced an LTD that was blocked by d-(-)-2-amino-5-phosphonopentanoic acid. Furthermore, an NMDA receptor-dependent depotentiation was induced in a time-dependent manner, being evoked by injection of CT 10 min, but not 1 hr, after LTP induction. These use- and time-dependent effects of the amyloidogenic peptides on synaptic plasticity promote long-lasting reductions in synaptic strength and oppose activity-dependent strengthening of transmission in the hippocampus. This will result in a profound disruption of information processing dependent on hippocampal synaptic plasticity.

A persistent reduction in short-term facilitation accompanies long-term potentiation in the CA1 area in the intact hippocampus.

Exploration of the nature of the relationship between short-term and long-term synaptic plasticity should aid our understanding of their roles in brain function. The effects of inducing long-term potentiation on short-term facilitation at CA1 synapses in the stratum radiatum of the intact hippocampus were examined by recording the slope of the field excitatory postsynaptic potential in both urethane and freely behaving adult rats. Facilitation of the second synaptic response to paired-pulse stimulation (40ms interstimulus interval) was monitored before and after the induction of long-term potentiation by high-frequency stimulation (10 trains of 20 pulses at 200Hz). The tetanus triggered a rapid overall reduction in paired-pulse facilitation that persisted for at least 2h. In the anaesthetized animals a detailed correlation analysis revealed that initial paired-pulse facilitation level correlated strongly with the subsequent reduction in paired-pulse facilitation and the magnitude of long-term potentiation. The reduction in paired-pulse facilitation also correlated with long-term potentiation magnitude. These relationships were not observed in animals with low initial degrees of paired-pulse facilitation. It was concluded that the relative contribution of different expression mechanisms of long-term potentiation varies depending on the initial facilitation characteristics of the synapses. Furthermore, the temporal selectivity and gain control of synapses can be altered persistently in the intact hippocampus. This suggests that there is considerable variation in the fidelity of temporal information storage at different synapses during learning and memory in the CA1 area.

Role of protein kinases A and C in the induction of mGluR-dependent long-term depression in the medial perforant path of the rat dentate gyrus in vitro.

The involvement of protein kinases A and C in the induction of low frequency stimulation-induced long-term depression (LTD) in the medial perforant path of the dentate gyrus in vitro has been studied using the selective PKA inhibitors H-89 and KT 5720 and PKC inhibitors Bisindolylmaleimide and Ro-31-8220. The PKC inhibitors Bisindolylmaleimide I and Ro-31-8220 and the PKA inhibitors H-89 and KT5720 all partially inhibited LTD induction. However, the presence of both a PKC and a PKA inhibitor was necessary to completely block LTD induction. The induction of long-term potentiation was not blocked by the inhibitors. It is suggested that the induction of LTD by LFS involves activation of PKC and PKA following activation of group I and group II metabotropic glutamate receptors (mGluR).

Induction of LTD by activation of group I mGluR in the dentate gyrus in vitro.

The ability of activation of group I metabotropic glutamate receptors (mGluR) to induce long-term depression (LTD) was investigated in the medial perforant path of the dentate gyrus in vitro. Application of the group I agonists (RS)-3,5-dihydroxyphenylglycine (DHPG) and (RS)-2-chloro-5-hydroxyphenylglycine (CHPG), and also the partial agonist (S)-(+)-2-(3'-Carboxybicyclo[1.1.1]pentyl)-glycine (UPF 596), induced LTD of the field EPSP. The induction of LTD is likely to be mediated via mGluR5 since CHPG and UPF 596 are selective agonists/partial agonists at that receptor. Further evidence for the involvement of group I mGluR in LTD induction was the finding, that the DHPG and low frequency stimulation induced LTD were inhibited by the group I mGluR antagonist [CRS]-1-aminoindan-1,5-dicarboxylic acid (AIDA). Investigation of the intracellular mechanisms underlying the induction of the group I mGluR-mediated LTD showed an inhibition of the LTD by the protein kinase C (PKC) inhibitor bisindolylmaleimide I and the protein tyrosine kinase inhibitor lavendustin A, but not the PKA inhibitor H89. These studies demonstrate that DHPG-induced LTD can be induced by the activation of mGluR5 followed by intracellular stimulation of PKC and tyrosine kinase.

Activation of mGluRII induces LTD via activation of protein kinase A and protein kinase C in the dentate gyrus of the hippocampus in vitro.

The involvement of metabotropic glutamate receptor group II (mGluRII) in the induction of long-term depression (LTD) was investigated in the medial perforant path of the rat dentate gyrus, a region with a very high density of mGluRII. Perfusion of either of two potent mGluRII agonists, (2S,1R,2R,3R)-2-(2S, 1'R, 2'R, 3'R)-2 (2' 3'-dicarboxycyclopropyl)glycine (DCG-IV) or (+)-2- aminobicyclo[3.1.0]hexane-2-6-dicarboxylic acid (LY354740) induced a reversible inhibition of the field EPSP followed, upon washout of the agonist, by LTD. The reversible inhibition was associated with a change in paired pulse depression, indicating an underlying presynaptic reduction in the probability of transmitter release, whereas the LTD was not associated with a change in paired pulse depression, indicating either a presynaptic reduction in the number of active release sites, or a postsynaptic change. Further evidence that the DCG-IV-induced LTD was generated by activation of mGluRII was the finding that the mGluRII antagonist (RS)-alpha-methylserine-O-phosphate monophenylphosphoryl ester (MSOPPE) prevented the induction of the LTD induced by DCG-IV. The DCG-IV-induced LTD showed mutual occlusion with LFS-induced LTD. The generation of the agonist-induced LTD required, in part, activation of N-methyl-D-aspartate receptors (NMDAR), as LTD induction was partially blocked in the presence of the NMDAR antagonist D-2-amino-5-phosphonopentanoate (AP5). Evidence for involvement of protein kinase C (PKC) and protein kinase (PKA) in the induction of LTD by activation of mGluRII was obtained by showing an inhibition of the DCG-IV-induced LTD by the PKC inhibitors Ro-31-8220 and bisindolylmaleimide I, and also by the PKA inhibitor H-89. The study demonstrates that activation of mGluRII induces LTD via activation the PKA and PKC pathways in the medial perforant path of the dentate gyrus.

Induction of long-lasting depression by (+)-alpha-methyl-4-carboxyphenylglycine and other group II mGlu receptor ligands in the dentate gyrus of the hippocampus in vitro.

Application of several well characterized group II mGlu receptor ligands was found to induce a long-lasting depression of synaptic transmission in the medial perforant path of the dentate gyrus. These ligands were N-acetylaspartylglutamate (NAAG), which is a dipeptide located in the brain and possibly functioning as a neurotransmitter, two agents widely used previously as mGluR antagonists, (+)-alpha-methyl-4-carboxyphenylglycine (MCPG), and (S)-alpha-ethylglutamate (EGLU), and the well characterized group II mGluR agonist (2S,1R,2R,3R)-2-(2S,1'R,2'R,3'R)-2(2'3'-dicarboxycyclopropyl)glyci ne (DCG-IV). It is postulated that all these ligands induced the long-lasting depression by an agonist/partial agonist action at group II mGlu receptor. The long-lasting depression induced by the ligands showed mutual occlusion with low frequency stimulation-induced long-term depression, demonstrating common induction or maintenance mechanisms. The induction of the long-lasting depression by the mGlu receptor ligands are suggested to occur postsynaptically as the induction was not associated with a change in paired pulse depression of excitatory postsynaptic potentials (EPSPs).