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Graham L Collingridge

Publications and source records attributed to Graham L Collingridge.

At least 37 records · Page 2Linked to original sources
PubMed2004

The GABA(B2) subunit is critical for the trafficking and function of native GABA(B) receptors.

Studies in heterologous systems have demonstrated that heterodimerisation of the two GABA(B) receptor subunits appears to be crucial for the trafficking and signalling of the receptor. Gene targeting of the GABA(B1) gene has demonstrated that the expression of GABA(B1) is essential for GABA(B) receptor function in the central nervous system (CNS). However, the contribution of the GABA(B2) subunit in the formation of native GABA(B) receptors is still unclear, in particular whether other proteins can substitute for this subunit. We have created a transgenic mouse in which the endogenous GABA(B2) gene has been mutated in order to express a C-terminally truncated version of the protein. As a result, the GABA(B1) subunit does not reach the cell surface and concomitantly both pre- and post-synaptic GABA(B) receptor functions are abolished. Taken together with previous gene deletion studies for the GABA(B1) subunit, this suggests that classical GABA(B) function in the brain is exclusively mediated by GABA(B1/2) heteromers.

Animals↗
PubMed2004

Endocannabinoids: losing inhibition to increase learning capacity?

Recent work has implicated endocannabinoids in various forms of synaptic plasticity. In this issue of Neuron, Chevaleyre and Castillo describe a new mechanism whereby a CB1 receptor-mediated LTD of inhibitory synaptic transmission facilitates the subsequent induction of LTP in a narrow band of synapses surrounding a region of potentiated synapses.

Animals↗
PubMed2004

Differential roles of NR2A and NR2B-containing NMDA receptors in cortical long-term potentiation and long-term depression.

It is widely believed that long-term depression (LTD) and its counterpart, long-term potentiation (LTP), involve mechanisms that are crucial for learning and memory. However, LTD is difficult to induce in adult cortex for reasons that are not known. Here we show that LTD can be readily induced in adult cortex by the activation of NMDA receptors (NMDARs), after inhibition of glutamate uptake. Interestingly there is no need to activate synaptic NMDARs to induce this LTD, suggesting that LTD is triggered primarily by extrasynaptic NMDA receptors. We also find that de novo LTD requires the activation of NR2B-containing NMDAR, whereas LTP requires activation of NR2A-containing NMDARs. Surprisingly another form of LTD, depotentiation, requires activation of NR2A-containing NMDARs. Therefore, NMDARs with different synaptic locations and subunit compositions are involved in various forms of synaptic plasticity in adult cortex.

2-Amino-5-phosphonovalerate↗
PubMed2004

Interactions between NMDA receptors and mGlu5 receptors expressed in HEK293 cells.

1 Ca2+ imaging was used to investigate interactions between responses induced by N-methyl-D-aspartate (NMDA; 15 microm) and (RS)-3,5-dihydroxyphenyl-glycine (DHPG; 30 microm) in human embryonic kidney (HEK) 293 cells, transiently transfected with rat recombinant NR1a, NR2A and mGlu5a cDNA. 2 Responses to NMDA were reversibly depressed by DHPG from 244+/-14 to 194+/-12% of baseline. Treatment with thapsigargin (1 microm, 10 min) prevented this effect. 3 After thapsigargin pretreatment, repeated applications of NMDA showed a gradual rundown in amplitude over a period of several hours, and were unaffected by DHPG. 4 Continuous perfusion with staurosporine (0.1 microm), after thapsigargin pretreatment, converted the run-down to a small increase in NMDA responses to 123+/-6 % of baseline. DHPG induced a further and sustained potentiation of NMDA responses to 174+/-12% of the initial baseline. 5 The protein tyrosine kinase (PTK) inhibitors genistein (50 microm) and 3-(4-chlorophenyl)1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (PP2; 1 microm) inhibited the staurosporine- and DHPG-induced potentiation of NMDA responses. 6 The protein phosphatase (PTP) inhibitors orthovanadate (100 microm) and phenyl arsine oxide (PAO, 1 microm) facilitated the staurosporine-evoked potentiation of NMDA responses and occluded DHPG-induced potentiation. 7 In conclusion, complex interactions can be demonstrated between mGlu5 and NMDA receptors expressed in HEK293 cells. There is a negative inhibitory influence of Ca2+ release and PKC activation. Inhibition of these processes reveals a tonic, mGlu5 receptor and PTK-dependent potentiation of NMDA receptors that can be augmented by either stimulating mGlu5 receptors or by inhibiting PTPs.

Animals↗
PubMed2004

Regulation of synaptic strength and AMPA receptor subunit composition by PICK1.

PICK1 (protein interacting with C kinase-1) regulates the surface expression of the AMPA receptor (AMPAR) GluR2 subunit, however, the functional consequences of this interaction are not well understood. Previous work has suggested that PICK1 promotes the internalization of AMPARs. However, we found that when PICK1 is virally expressed in the CA1 region of hippocampal slices, it causes an increase in AMPAR-mediated EPSC amplitude. This effect is associated with increased AMPAR rectification and sensitivity to polyamine toxin. These effects are blocked by PKC or calcium/calmodulin-dependent protein kinase II inhibitors, indicating that the virally expressed PICK1 signals through an endogenous kinase cascade. In contrast, blockade of interactions with GluR2 at the N-ethylmaleimide-sensitive factor site did not cause a change in subunit composition, suggesting that the effects of PICK1 are not simply a nonspecific consequence of removing AMPARs from the surface. Immunocytochemical and biochemical analyses in dissociated cultured hippocampal neurons show that PICK1 causes a decrease in endogenous GluR2 surface expression but no change in GluR1 surface levels. To address the physiological role of PICK1, we virally expressed C-terminal GluR2 peptides. Blockade of endogenous PICK1 PDZ (postsynaptic density-95/Discs large/zona occludens-1) domain interactions produced opposite effects on synaptic strength and AMPAR rectification to those observed with PICK1 expression. This demonstrates that AMPAR subunit composition is physiologically regulated through a mechanism involving PICK1 PDZ domain interactions. These findings suggest that PICK1 acts to downregulate the GluR2 content of AMPARs at hippocampal CA1 synapses, thereby increasing synaptic strength at resting membrane potentials.

Animals↗
PubMed2004

Removal of AMPA receptors (AMPARs) from synapses is preceded by transient endocytosis of extrasynaptic AMPARs.

AMPA receptors (AMPARs) are dynamically regulated at synapses, but the time course and location of their exocytosis and endocytosis are not known. Therefore, we have used ecliptic pHluorin-tagged glutamate receptor 2 to visualize changes in AMPAR surface expression in real time. We show that synaptic and extrasynaptic AMPARs respond very differently to NMDA receptor activation; there is a rapid internalization of extrasynaptic AMPARs that precedes the delayed removal of synaptic AMPARs.

Animals↗
PubMed2004

Multiple, developmentally regulated expression mechanisms of long-term potentiation at CA1 synapses.

Long-term potentiation (LTP) of AMPA receptor-mediated synaptic transmission at hippocampal CA1 synapses has been extensively studied, but the mechanisms responsible for its expression remain unresolved. We tested a hypothesis that there are multiple, developmentally regulated expression mechanisms by directly comparing LTP in hippocampal slices obtained from rats of two ages. At postnatal day 12 (P12), LTP was fully accounted for by an increase in potency (mean amplitude of responses excluding failures). This was associated with either an increase in AMPA receptor single-channel conductance (gamma) or no change in gamma, suggesting an increase in the number of AMPA receptors. At P6, LTP was explained by an additional two mechanisms. In the majority of neurons, LTP was associated with an increase in success rate and a decrease in paired-pulse facilitation. In the remaining neurons, LTP was attributable to an increase in potency. However, in contrast to P12 neurons, the potency increase was associated with a decrease in gamma, suggesting the insertion of receptors with lower gamma. We conclude that there are multiple expression mechanisms for LTP at CA1 synapses that are developmentally regulated. These findings suggest that a single class of synapse uses a number of different molecular mechanisms to produce long-term changes in synaptic strength.

Animals↗
PubMed2004

Characterisation of UBP296: a novel, potent and selective kainate receptor antagonist.

Willardiine derivatives with an N3-benzyl substituent bearing an acidic group have been synthesized with the aim of producing selective antagonists for GLUK5-containing kainate receptors. UBP296 was found to be a potent and selective antagonist of native GLUK5-containing kainate receptors in the spinal cord, with activity residing in the S enantiomer (UBP302). In cells expressing human kainate receptor subunits, UBP296 selectively depressed glutamate-induced calcium influx in cells containing GLUK5 in homomeric or heteromeric forms. In radioligand displacement binding studies, the willardiine analogues displaced [3H]kainate binding with IC50 values >100 microM at rat GLUK6, GLUK2 or GLUK6/GLUK2. An explanation of the GLUK5 selectivity of UBP296 was obtained using homology models of the antagonist bound forms of GLUK5 and GLUK6. In rat hippocampal slices, UBP296 reversibly blocked ATPA-induced depressions of synaptic transmission at concentrations subthreshold for affecting AMPA receptor-mediated synaptic transmission directly. UBP296 also completely blocked the induction of mossy fibre LTP, in medium containing 2 mM (but not 4 mM) Ca2+. These data provide further evidence for a role for GLUK5-containing kainate receptors in mossy fibre LTP. In conclusion, UBP296 is the most potent and selective antagonist of GLUK5-containing kainate receptors so far described.

Alanine↗
PubMed2004

Receptor trafficking and synaptic plasticity.

Long-term potentiation and long-term depression are processes that have been widely studied to understand the molecular basis of information storage in the brain. Glutamate receptors are required for the induction and expression of these forms of plasticity, and GABA (gamma-aminobutyric acid) receptors are involved in their modulation. Recent insights into how these receptors are rapidly moved into and out of synaptic membranes has profound implications for our understanding of the mechanisms of long-term potentiation and long-term depression.

Animals↗
PubMed2003

A role for Ca2+ stores in kainate receptor-dependent synaptic facilitation and LTP at mossy fiber synapses in the hippocampus.

Compared with NMDA receptor-dependent LTP, much less is known about the mechanism of induction of NMDA receptor-independent LTP; the most extensively studied form of which is mossy fiber LTP in the hippocampus. In the present study we show that Ca2+-induced Ca2+ release from intracellular stores is involved in the induction of mossy fiber LTP. This release also contributes to the kainate receptor-dependent component of the pronounced synaptic facilitation that occurs during high-frequency stimulation. We also present evidence that the trigger for this Ca2+ release is Ca2+ permeation through kainate receptors. However, these novel synaptic mechanisms can be bypassed when the Ca2+ concentration is raised (from 2 to 4 mM), via a compensatory involvement of L-type Ca2+ channels. These findings suggest that presynaptic kainate receptors at mossy fiber synapses can initiate a cascade involving Ca2+ release from intracellular stores that is important in both short-term and long-term plasticity.

2-Amino-5-phosphonovalerate↗
PubMed2003

Kainate receptors and the induction of mossy fibre long-term potentiation.

There is intense interest in understanding the molecular mechanisms involved in long-term potentiation (LTP) in the hippocampus. Significant progress in our understanding of LTP has followed from studies of glutamate receptors, of which there are four main subtypes (alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), N-methyl-D-aspartate (NMDA), mGlu and kainate). This article summarizes the evidence that the kainate subtype of glutamate receptor is an important trigger for the induction of LTP at mossy fibre synapses in the CA3 region of the hippocampus. The pharmacology of the first selective kainate receptor antagonists, in particular the GLU(K5) subunit selective antagonist LY382884, is described. LY382884 selectively blocks the induction of mossy fibre LTP, in response to a variety of different high-frequency stimulation protocols. This antagonist also inhibits the pronounced synaptic facilitation of mossy fibre transmission that occurs during high-frequency stimulation. These effects are attributed to the presence of presynaptic GLU(K5)-subunit-containing kainate receptors at mossy fibre synapses. Differences in kainate receptor-dependent synaptic facilitation of AMPA and NMDA receptor-mediated synaptic transmission are described. These data are discussed in the context of earlier reports that glutamate receptors are not involved in mossy fibre LTP and more recent experiments using kainate receptor knockout mice, that argue for the involvement of GLU(K6) but not GLU(K5) kainate receptor subunits. We conclude that activation of presynaptic GLU(K5)-containing kainate receptors is an important trigger for the induction of mossy fibre LTP in the hippocampus.

Animals↗
PubMed2003

Hippocampal synaptic plasticity in mice carrying the rd mutation in the gene encoding cGMP phosphodiesterase type 6 (PDE6).

Cyclic GMP (cGMP) has been implicated in the modulation of long-term potentiation (LTP) and depression (LTD) in the hippocampus. Transcripts for subunits of several types of cGMP specific phosphodiesterase are found in the mammalian brain but their relative role in hippocampal function is unclear. The retinal degeneration (rd) mutation in the gene encoding the PDE6B subunit causes a loss of function in PDE6 enzyme and in adult mice homozygous to the mutation it causes blindness. We have used this natural mutation, and the cGMP phosphodiesterase inhibitor zaprinast, in wild-type and rd/rd mouse littermates to investigate whether PDE5 and/or PDE6 regulates excitatory synaptic transmission in the hippocampus. Mice were genotyped using two independent PCR methods. Glutamate-mediated synaptic transmission in the CA1 region or dentate gyrus was unaffected in hippocampal brain slices from mice carrying the rd mutation. Similarly the facilitation of synaptic events by paired-pulse stimuli, and LTP induced by a theta-burst (10 bursts of four events at 100 Hz with a 200-ms inter-burst interval) were normal in rd/rd mice. Inhibition of cGMP-specific PDE activity by zaprinast (10 microM, an inhibitor of PDE5 and PDE6) induced a slowly developing and sustained depression of field synaptic potentials that was quantitatively similar in both wild-type and rd/rd mice. Thus in the CA1 region synaptic plasticity is likely to be regulated by the PDE5 rather than the PDE6 isoform.

Animals↗
PubMed2003

Rapid and differential regulation of AMPA and kainate receptors at hippocampal mossy fibre synapses by PICK1 and GRIP.

We identified four PDZ domain-containing proteins, syntenin, PICK1, GRIP, and PSD95, as interactors with the kainate receptor (KAR) subunits GluR5(2b,) GluR5(2c), and GluR6. Of these, we show that both GRIP and PICK1 interactions are required to maintain KAR-mediated synaptic function at mossy fiber-CA3 synapses. In addition, PKC alpha can phosphorylate ct-GluR5(2b) at residues S880 and S886, and PKC activity is required to maintain KAR-mediated synaptic responses. We propose that PICK1 targets PKC alpha to phosphorylate KARs, causing their stabilization at the synapse by an interaction with GRIP. Importantly, this mechanism is not involved in the constitutive recycling of AMPA receptors since blockade of PDZ interactions can simultaneously increase AMPAR- and decrease KAR-mediated synaptic transmission at the same population of synapses.

Adaptor Proteins, Signal Transducing↗
PubMed2003

Activation of mGlu receptors induces LTD without affecting postsynaptic sensitivity of CA1 neurons in rat hippocampal slices.

Two forms of long-term depression (LTD) of excitatory synaptic transmission have been identified in the mammalian CNS, which are induced by the synaptic activation of N-methyl-D-aspartate (NMDA) and metabotropic glutamate (mGlu) receptors, respectively. The mGlu receptor-dependent form of LTD can be activated by application of 3,5-dihydroxyphenylglycine (DHPG), a group I selective mGlu receptor agonist. DHPG-induced LTD is increasingly being used to investigate the mechanisms of mGlu receptor-dependent LTD. However, recent experiments have argued for both a pre- and postsynaptic locus of expression of DHPG-induced LTD. In the present study we report that DHPG-induced LTD is not associated with changes in the sensitivity of CA1 neurons to bath applied AMPA. Furthermore, in contrast to homosynaptic LTD, DHPG-induced LTD is also not associated with changes in sensitivity to focally uncaged L-glutamate. These data do not support the notion that DHPG-induced LTD requires a modification of AMPA receptors, such as their internalisation, but are compatible with a presynaptic mechanism of expression.

Animals↗
PubMed2003

Mechanisms contributing to the exacerbated epileptiform activity in hippocampal slices of GABAB1 receptor subunit knockout mice.

The recently developed GABAB1 receptor subunit knockout (GABAB1 -/-) mouse displays complete loss of GABAB receptor function and develops complex generalized epilepsies including absence type, audiogenic as well as spontaneous generalized seizures with electrographic spike-wave discharge signatures. To gain insight into the cellular mechanisms contributing to the generation and maintenance of this epileptic phenotype we have compared epileptiform activity induced in hippocampal slices obtained from GABAB1 -/- and wild type (GABAB1 +/+) littermates. Deletion of the GABAB1 receptor subunit had no effect on a range of passive membrane properties of CA3 pyramidale neurones, non-synaptic epileptiform field bursting and spreading depression recorded in 6mM K+/Ca2+-free medium, and inter-ictal synaptically-induced epileptiform activity induced by 100 microM 4-aminopyridine (4-AP). In contrast, synaptic epileptiform activity induced by 10 microM bicuculline, removal of extracellular Mg2+ or addition of 10 microM oxotremorine was enhanced in GABAB1 -/- slices. Acute blockade of GABAB receptors using a selective antagonist only partly mimicked these effects. It is suggested that the exaggerated in vitro epileptiform activity is caused by both acute and chronic consequences of the loss of GABAB receptor function in vivo. Specifically, enhancement of N-methyl-d-aspartate (NMDA) receptor triggered synaptic processes, arising from the loss of the GABAB receptor-mediated inhibitory postsynaptic potential (IPSP, together with a possible promotion of depolarising IPSPs due to the removal of GABAB autoreceptor function) is likely to underlie these effects.

Animals↗
PubMed2003

Parallel kinase cascades are involved in the induction of LTP at hippocampal CA1 synapses.

To identify the enzymes involved in the induction of N-methyl-D-aspartate (NMDA) receptor-dependent long-term potentiation (LTP) at CA1 synapses of two-week-old rats we have tested various kinase inhibitors. Surprisingly, given the large body of evidence supporting a role for calcium/calmodulin-dependent protein kinase II (CaMKII) in LTP, inhibition of this enzyme did not affect the induction of LTP at this age. Similarly inhibition of protein kinase A (PKA) or protein kinase C (PKC) was also without effect. However, inhibition of CaMKII together with inhibition of either PKA or PKC fully blocked the induction of LTP. These experiments reveal, unexpectedly, the existence of two parallel kinase pathways, one involving CaMKII and the other PKA and PKC, either of which can fully support the induction of LTP, at this stage of development.

Animals↗