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M Baudry

Publications and source records attributed to M Baudry.

At least 55 records · Page 3Linked to original sources
PubMed1998

KID-1, a protein kinase induced by depolarization in brain.

Membrane depolarization leads to changes in gene expression that modulate neuronal plasticity. Using representational difference analysis, we have identified a previously undiscovered cDNA, KID-1 (kinase induced by depolarization), that is induced by membrane depolarization or forskolin, but not by neurotrophins or growth factors, in PC12 pheochromocytoma cells. KID-1 is an immediate early gene that shares a high degree of sequence similarity with the family of PIM-1 serine/threonine protein kinases. Recombinant KID-1 fusion protein is able to catalyze both histone phosphorylation and autophosphorylation. KID-1 mRNA is present in a number of unstimulated tissues, including brain. In response to kainic acid and electroconvulsive shock-induced seizures, KID-1 is induced in specific regions of the hippocampus and cortex.

Amino Acid Sequence↗
PubMed1998

Phosphorylation regulates calpain-mediated truncation of glutamate ionotropic receptors.

Pre-incubation of synaptic membranes with phosphatase inhibitors significantly reduces the extent of calpain-mediated truncation of both GluR1 and NR2 subunits of AMPA and NMDA receptors, respectively. The same treatment did not modify calpain-mediated truncation of spectrin. These results might have important implications for mechanisms of synaptic plasticity as the balance of kinase/phosphatase activity and calpain has been proposed to regulate synaptic efficacy at glutamatergic synapses.

Animals↗
PubMed1998

The tight association of protein kinase CK2 with plasma membranes is mediated by a specific domain of its regulatory beta-subunit.

Previous immunocytochemical studies have shown that protein kinase CK2 is mostly detected both in the cytoplasm and the nucleus of most cells. In the present study, CK2 was detected in highly purified plasma membrane preparations from rat liver. The protein kinase could be released from the membranes by high salt extraction (>1 M NaCl). Plasma membranes prepared from SF9 insect cells expressing the alpha- and beta-subunits of CK2 also contained a significant amount of oligomeric CK2. Furthermore, it was demonstrated in this cell system as well as in rat liver plasma membranes, that the beta-subunit of the kinase is the targeting subunit which mediates the tight association of the enzyme to plasma membrane components. Binding studies using membranes and recombinant proteins corresponding to different regions of the beta-subunit suggest that a functional domain previously shown to be involved in the binding of polyamines may also participate to the binding of CK2 to membranes. Modification of membranes by trypsin and phospholipases indicated that the binding process may require both membrane protein(s) and phospholipids. Interestingly, it was observed that the amount of membrane-bound CK2 in liver of embryos and new born rats increases dramatically after birth and persists during the postnatal stages of development.

Animals↗
PubMed1998

Developmental changes in subcellular AMPA/GluR receptor populations in rat forebrain.

Forebrains from rats of postnatal days (PND) 2, 7, 14, 21, and 30-40 were subjected to subcellular fractionation and samples from crude mitochondrial (P2, which contain synaptic plasma membranes) and microsomal (P3) fractions were used for SDS-PAGE and Western blotting with antibodies against GluR1, and GluR2/3 subunits of AMPA/GluR receptors. GluR immunoreactivity in P2 fractions increased gradually from PND 2 to PND 30. In contrast, GluR immunoreactivity in P3 fractions increased sharply at early postnatal ages, and was higher than in adults as early as at PND 7. Data were compared to postnatal changes in 3H-AMPA binding reported in various studies. Significant correlations were observed between changes in GluR immunoreactivity in P3 fractions and changes in high-affinity binding on one hand and between changes in GluR immunoreactivity in P2 fractions, and changes in low affinity binding. These data further establish that glutamate receptors present in different subcellular compartments represent different maturational states of the receptors, and suggest that changes in GluR populations could participate in mechanisms of synaptic plasticity.

Animals↗
PubMed1998

Calpain-mediated regulation of NMDA receptor structure and function.

Calpains have been previously shown to regulate AMPA receptor properties by producing partial truncation of the C-terminal domains of several receptor subunits. We now report that NMDA receptor subunits, in particular NR2 subunits, are also subjected to calpain-mediated truncation. Treatment of synaptic membranes with calpain I resulted in truncation of both NR1 and NR2 subunits, with the appearance of NR2 species with lower mol.wt. than native subunits, but still recognized by antibodies directed at the C-terminal domain. This treatment did not modify the binding of several ligands of the NMDA receptors, such as glutamate, glycine or TCP. Incubation of thin frozen-thawed brain sections with calcium resulted in calpain-mediated selective degradation of NR2 subunits, as truncation into smaller fragments was totally blocked by calpain inhibitors. Under the same conditions, TCP binding to sections was decreased by about 50%, an effect also blocked by calpain inhibitors. Treatment of hippocampal slices in culture with the excitotoxin, kainic acid, also produced calpain-mediated truncation of the C-terminal domain of NR2 but not NR1 subunits of the NMDA receptors. The results indicate that calpain activation produces several modifications of NMDA receptors, including the truncation of the C-terminal domain of NR2 subunits, and changes in channel binding properties. They suggest that calpain-mediated regulation of NMDA receptors might represent a feed-back regulation of the receptors which could be used to limit receptor activation.

Animals↗
PubMed1998

Calpain-mediated proteolysis of GluR1 subunits in organotypic hippocampal cultures following kainic acid treatment.

The present study examined changes in GluR1 subunits after kainic acid (KA) treatment of organotypic hippocampal cultures. Immunoblots labeled with antibodies directed at the C-terminal domain of GluR1 revealed a large decrease in GluR1 immunoreactivity at 6 h and 24 h, while immunoblots labeled with antibodies directed at the N-terminal domain indicated that KA treatment produced changes in structure but not amount of GluR1 protein. Changes in GluR1 subunits were significantly reduced by the calpain inhibitor, calpeptin. These results indicate that KA-induced changes in GluR1 properties are mediated by calpain activation.

Calpain↗
PubMed1998

Bidirectional modulation of AMPA receptor properties by exogenous phospholipase A2 in the hippocampus.

The synaptic modifications underlying long-term potentiation (LTP) and long-term depression (LTD) of synaptic transmission in various brain structures may result from changes in the properties of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) subtype of glutamate receptors. In the present study, we report that treatment of rat synaptoneurosomes with increasing concentrations of phospholipase A2 (PLA2) produces a biphasic effect on AMPA receptor binding, with low concentrations causing a decrease and high concentrations an increase in agonist binding. Analysis of the saturation kinetics of 3H-AMPA binding revealed that the biphasic effect of PLA2 was due to modifications in receptor affinity and not to changes in the maximum number of binding sites for AMPA receptors. The 12-lipoxygenase inhibitors preferentially reduced PLA2-induced decrease in AMPA binding and treatment of hippocampal synaptoneurosomes with arachidonic acid (AA) or 12-HPETE, the first metabolite generated from the hydrolysis of AA by 12-lipoxygenases, decreased 3H-AMPA binding. Moreover, electrophysiological experiments indicated that the 12-lipoxygenase inhibitor baicalein totally blocked LTD formation in area CA1 of hippocampal slices. The decrease in 3H-AMPA binding elicited by low concentrations of PLA2, as well as the level of LTD, were partially reduced by AA-861, a 5-lipoxygenase inhibitor, while the cyclooxygenase inhibitor indomethacin did not prevent LTD formation or the effects of PLA2 on 3H-AMPA binding. Our results provide evidence for a possible involvement of lipoxygenase metabolites in the regulation of AMPA receptor during synaptic depression. In addition, they strongly support the idea that the same biochemical pathway, i.e., NMDA receptor activation and endogenous PLA2 stimulation, may represent a common mechanism resulting in AMPA receptor alterations for both LTP and LTD formation.

Animals↗
PubMed1998

Rapid effects of kainate administration on alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor properties in rat hippocampus.

We investigated changes in AMPA receptor properties in rat hippocampus 5 h after systemic kainate administration. Quantitative [3H]AMPA autoradiography and Western blot analysis of receptor subunits GluR1-3 in different subcellular fractions were used to evaluate possible alterations in binding characteristics and immunological properties of the receptors in synaptic and nonsynaptic fractions. Both ligand-binding and Western blots revealed significant changes in binding and immunological properties of nonsynaptic receptors but relatively smaller changes in synaptic receptors 5 h after kainate administration. GluR2/3 showed a greater relative change in the synaptic receptor population compared to GluR1, suggesting either a shift in subunit composition of AMPA receptors or the formation of a synaptic subpopulation of AMPA receptors with truncated C-terminal domain of GluR1 subunits. The effects of kainic acid were blocked by cycloheximide treatment indicating that the changes were due at least in part to increased synthesis of AMPA receptor subunits. The results indicate that excessive synaptic activity produces rapid changes in both synaptic and nonsynaptic AMPA receptor properties.

Analysis of Variance↗
PubMed1998

Introduction

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Journal Article↗
PubMed1998

The narrow band hypothesis: an interesting approach for high-intensity transient signals (HITS) detection.

We propose a new approach to detect microemboli automatically using the narrow band hypothesis. An initial database of 560 peripheral arterial Doppler high-intensity transient signals (HITS) was created to study microemboli and to define the normal limits to be used in our method. When a HITS occurs, our approach consists of modelling the Doppler signal using amplitude and frequency wave modulation. A threshold was defined experimentally using this database and then applied to 38 recordings from 12 patients. Using another database, six expert Doppler users reported 140, 176, 155, 161, 161 and 146 HITS, corresponding to a total of 197 different observed HITS. When an event was detected by 6, 5, 4, 3, 2 and 1 of the observers, the sensitivity of the automatic detection was 94.8%, 75.9%, 55.6%, 42.9%, 30% and 0%, respectively. The sensitivity of our automatic detection thus is highly associated with the likelihood (defined as the ratio of observers in agreement to the total number of observers) of an event: r = 0.99 for p < 0.0001. Although future research would result in improvement of the specificity, the narrow band hypothesis appears to be a promising technique for the detection of HITS.

Blood Flow Velocity↗
PubMed1998

Exposure to a conditioned aversive environment interferes with long-term potentiation induction in the fimbria-CA3 pathway.

The effect of re-exposure of rats to an aversive environment on the induction of long-term potentiation was investigated in the CA3 region 3 and 12 h after contextual conditioning. Electro-physiological recordings showed that re-exposure of rats to the conditioning chamber produced a significant and long-lasting decrease in population spike amplitude at both post-conditioning delays. High-frequency stimulation of the fimbria induced a large and persistent increase in CA3 population spike amplitude (about 400% of baseline) in animals of control groups and shocked animals that were not re-exposed to the conditioning environment. However, high-frequency stimulation applied during re-exposure of shocked subjects 3 h after the initial exposure resulted in a small and transient increase in population spike amplitude (about 140% of baseline); when applied 12 h after the initial exposure, it produced a persistent depression of the response (-30% of baseline). Behavioural testing indicated that re-exposure of shocked animals to the conditioning environment elicited a qualitatively and quantitatively similar freezing behaviour at both post-conditioning delays (3 or 12 h). In contrast to the long-lasting decrease in CA3 population spike amplitude produced by re-exposure to the aversive environment, the level of freezing behaviour diminished rapidly within 10 min of exposure. These results suggest that, during exposure to a conditioned aversive environment, alterations in fimbria-CA3 neural processing may be dissociated from contextual fear-induced freezing behaviour. In addition, processes underlying long-term potentiation induction in fimbria-CA3 pathway may be opposite to those taking place during hippocampal processing of conditioned aversive contexts.

Animals↗
PubMed1998

High- and low-affinity alpha-[3H]amino-3-hydroxy-5-methylisoxazole-4-propionic acid ([3H]AMPA) binding sites represent immature and mature forms of AMPA receptors and are composed of differentially glycosylated subunits.

Quantitative alpha-[3H]amino-3-hydroxy-5-methylisoxazole-4-propionic acid ([3H]AMPA) binding autoradiography was performed on frozen-thawed sections from rat brain after preincubation at 0 or 35 degrees C for 1 h. Preincubation at 35 degrees C instead of 0 degrees C resulted in a selective decrease of [3H]AMPA binding assayed at a low concentration of [3H]AMPA (50 nM) and an enhancement of binding at a high concentration (500 nM). The decrease in [3H]AMPA binding after preincubation at 35 degrees C was accompanied with the loss of the lighter organelles of P3 (microsomal) fractions. These organelles were found to contain a small subpopulation of AMPA/GluR receptors exhibiting a high affinity for [3H]AMPA (K(D) approximately 14 nM), whereas heavier organelles exhibited lower affinity for AMPA (K(D) approximately 190 nM). This small subpopulation of AMPA/GluR receptors contained almost exclusively a structurally distinct species of GluR2/3 subunits with an apparent molecular mass of 103.5 kDa (assessed with anti-GluR2/3, C-terminal antibodies). Experiments using two deglycosylating enzymes, N-glycopeptidase F and endoglycosidase H, clearly indicated that the 103.5-kDa species represented a partially unglycosylated form of GluR2/3 subunits containing the high-mannose type of oligosaccharide moiety, whereas receptors present in synaptosomal fractions were composed of subunits with complex oligosaccharides. A similar result was obtained by using an antibody recognizing the N-terminal domain of GluR2(4). The same enzymatic treatment indicated that GluR1 subunits also exhibited a partially glycosylated form. These data indicate that high-affinity [3H]AMPA binding sites represent nonsynaptic, intracellular membrane-bound AMPA receptors that differ from synaptic receptors by at least the glycosylation state of GluR2 (and GluR1) subunits. In addition, our results provide a relatively simple way of assessing changes in two spatially and structurally distinct [3H]AMPA binding/GluR sites.

Amidohydrolases↗
PubMed1997

Effects of a nitric oxide synthase inhibitor on NMDA receptor function in organotypic hippocampal cultures.

Nitric oxide (NO) has been proposed to trigger long-term potentiation (LTP) at CA3 to CA1 synapses. We previously reported that NO synthesis inhibitors and blockers reduce an electrophysiological index of NMDA receptor activation in acute hippocampal slices. We now show that the NOS inhibitor, NG-methyl-L-arginine (MLA), also reversibly prevents LTP induction in organotypic hippocampal slices and significantly reduces a biochemical index of NMDA receptor function. These results results further indicate that MLA inhibits LTP induction by interfering with NMDA receptor functions.

Animals↗
PubMed1997

Time-dependent blockade of STP and LTP in hippocampal slices following acute stress in mice.

The characteristics of short-term potentiation (STP) and long-term potentiation (LTP) in the CA1 region of hippocampal slices were determined at various times following exposure to acute stress produced by restraint and tail-shock in mice. In slices prepared from control animals, theta-burst stimulation resulted in a large increase in evoked field excitatory postsynaptic potentials (EPSPs) amplitude and slope that remained stable at least up to 30 min after stimulation. Slices prepared 1 h after stress exhibited a marked decrease in the extent of both STP and LTP. STP and LTP magnitude were still significantly decreased 24 h after stress exposure and were completely restored to control levels by 48 h. These results provide evidence for a reversible impairment of STP and LTP in CA1 following an acute episode of stress, and suggest that stress activates processes different from those activated by LTP-inducing stimuli.

Animals↗
PubMed1997

Glycine-induced long-term potentiation is associated with structural and functional modifications of alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid receptors.

Global long-term potentiation (LTP) was induced in organotypic hippocampal slice cultures by a brief application of 10 mM glycine. Glycine-induced LTP was occluded by previous theta burst stimulation-induced potentiation, indicating that both phenomena share similar cellular processes. Glycine-induced LTP was associated with increased [3H]alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA) binding in membrane fractions as well as increased amount of a selective spectrin breakdown product generated by calpain-mediated spectrin proteolysis. Antibodies against the C-terminal (C-Ab) and N-terminal (N-Ab) domains of GluR1 subunits were used to evaluate structural changes in AMPA receptor properties resulting from glycine-induced LTP. No quantitative or qualitative changes were observed in Western blots from membrane fractions prepared from glycine-treated slices with C-Ab. In contrast, Western blots stained with N-Ab revealed the formation of a 98-kDa species of GluR1 subunits as well as an increased amount of immunoreactivity after glycine-induced LTP. The amount of spectrin breakdown product was positively correlated with the amount of the 98-kDa species of GluR1 after glycine treatment. Functional modifications of AMPA receptors were evaluated by determining changes in the effect of pressure-applied AMPA on synaptic responses before and after glycine-induced LTP. Glycine treatment produced a significant increase in AMPA receptor function after potentiation that correlated with the degree of potentiation. The results indicate that LTP induction produces calpain activation, truncation of the C-Ab domain of GluR1 subunits of AMPA receptors, and increased AMPA receptor function. They also suggest that insertion of new receptors takes place after LTP induction.

Animals↗
PubMed1997

Novel expression mechanism for synaptic potentiation: alignment of presynaptic release site and postsynaptic receptor.

A combination of experimental and modeling approaches was used to study cellular-molecular mechanisms underlying the expression of short-term potentiation (STP) and long-term potentiation (LTP) of glutamatergic synaptic transmission in the hippocampal slice. Electrophysiological recordings from dentate granule cells revealed that high-frequency stimulation of perforant path afferents induced a robust STP and LTP of both (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and N-methyl-D-aspartic acid (NMDA) receptor-mediated synaptic responses. However, the decay time constant for STP of the AMPA receptor-mediated excitatory postsynaptic potential was approximately 6 min, whereas the decay time constant for STP of the NMDA receptor-mediated excitatory postsynaptic potential was only 1 min. In addition, focal application of agonists during the expression of STP revealed that the magnitude of conductance change elicited by NMDA application was significantly enhanced, whereas the magnitude of conductance change elicited by application of AMPA remained constant. These findings are most consistent with a postsynaptic mechanism of STP and LTP. Different putative mechanisms were evaluated formally using a computational model that included diffusion of glutamate within the synaptic cleft, different kinetic properties of AMPA and NMDA receptor/channels, and geometric relations between presynaptic release sites and postsynaptic receptor/channels. Simulation results revealed that the only hypothesis consistent with experimental data is that STP and LTP reflect a relocation of AMPA receptor/channels in the postsynaptic membrane such that they become more closely "aligned" with presynaptic release sites. The same mechanism cannot account for STP or LTP of NMDA receptor-mediated responses; instead, potentiation of the NMDA receptor subtype is most consistent with an increase in receptor sensitivity or number.

Animals↗
PubMed1997

Induction of tumor suppressor p53 and DNA fragmentation in organotypic hippocampal cultures following excitotoxin treatment.

The p53 tumor suppressor gene encodes a cell cycle regulatory protein that is induced by DNA damage and has been implicated in apoptosis. To investigate whether excitotoxic cell death due to kainic acid (KA) and cell death due to N-methyl-D-aspartate (NMDA) share similar molecular mechanisms, we studied p53 expression and DNA fragmentation in organotypic hippocampal slice cultures following excitotoxin treatment. Cellular analyses showed that both p53 induction and DNA fragmentation occurred only in injured neurons following exposure to either excitotoxin. The temporal profiles of these changes demonstrated that p53 induction preceded DNA fragmentation. The extent of regional alterations in p53 expression and DNA fragmentation correlated with drug-related toxicity (i.e., NMDA > KA). These results support the hypothesis that p53 is a marker of neuronal death in the CNS and suggest the possibility that excitotoxin-mediated neuronal death may occur through a p53-dependent pathway.

Animals↗