Biomedical subjects

Laurent Groc

Publications and source records attributed to Laurent Groc.

15 recordsLinked to original sources

PubMed2006

NMDA receptor surface mobility depends on NR2A-2B subunits.

The NR2 subunit composition of NMDA receptors (NMDARs) varies during development, and this change is important in NMDAR-dependent signaling. In particular, synaptic NMDAR switch from containing mostly NR2B subunit to a mixture of NR2B and NR2A subunits. The pathways by which neurons differentially traffic NR2A- and NR2B-containing NMDARs are poorly understood. Using single-particle and -molecule approaches and specific antibodies directed against NR2A and NR2B extracellular epitopes, we investigated the surface mobility of native NR2A and NR2B subunits at the surface of cultured neurons. The surface mobility of NMDARs depends on the NR2 subunit subtype, with NR2A-containing NMDARs being more stable than NR2B-containing ones, and NR2A subunit overexpression stabilizes surface NR2B-containing NMDARs. The developmental change in the synaptic surface content of NR2A and NR2B subunits was correlated with a developmental change in the time spent by the subunits within synapses. This suggests that the switch in synaptic NMDAR subtypes depends on the regulation of the receptor surface trafficking.

Animals↗

PubMed2006

Single nanoparticle photothermal tracking (SNaPT) of 5-nm gold beads in live cells.

Tracking individual nano-objects in live cells during arbitrary long times is a ubiquitous need in modern biology. We present here a method for tracking individual 5-nm gold nanoparticles on live cells. It relies on the photothermal effect and the detection of the Laser Induced Scattering around a NanoAbsorber (LISNA). The key point for recording trajectories at video rate is the use of a triangulation procedure. The effectiveness of the method is tested against single fluorescent molecule tracking in live COS7 cells on subsecond timescales. We further demonstrate recordings for several minutes of AMPA receptors trajectories on the plasma membrane of live neurons. Single Nanoparticle Photothermal Tracking has the unique potential to record arbitrary long trajectory of membrane proteins using nonfluorescent nanometer-sized labels.

Animals↗

PubMed2006

AMPA and NMDA glutamate receptor trafficking: multiple roads for reaching and leaving the synapse.

Glutamate receptor trafficking in and out of synapses is one of the core mechanisms for rapid changes in the number of functional receptors during synaptic plasticity. Recent data have shown that the fast gain and loss of receptors from synaptic sites are accounted for by endocytic/exocytic processes and by their lateral diffusion in the plane of the membrane. These events are interdependent and regulated by neuronal activity and interactions with scaffolding proteins. We review here the main cellular steps for AMPA and NMDA receptor synthesis, traffic within intracellular organelles, membrane exocytosis/endocytosis and surface trafficking. We focus on new findings that shed light on the regulation of receptor cycling events and surface trafficking and the way that this might reshape our thinking about the specific regulation of receptor accumulation at synapses.

Animals↗

PubMed2006

Multiple routes for glutamate receptor trafficking: surface diffusion and membrane traffic cooperate to bring receptors to synapses.

Trafficking of glutamate receptors into and out of synapses is critically involved in the plasticity of excitatory synaptic transmission. Endocytosis and exocytosis of receptors have initially been thought to account alone for this trafficking. However, membrane proteins also traffic through surface lateral diffusion in the plasma membrane. We describe developments in electrophysiological and optical approaches that have allowed for the real-time measurement of glutamate receptor surface trafficking in live neurons. These include (i) specific imaging of surface receptors using a pH-sensitive fluorescent protein; (ii) design of a photoactivable drug to locally inactivate surface receptors and monitor electrophysiologically their recovery; and (iii) application of single-molecule fluorescence microscopy to directly track the movement of individual surface receptors with nanometer resolution inside and outside synapses. Together, these approaches have demonstrated that glutamate receptors diffuse at high rates in the neuronal membrane and suggest a key role for surface diffusion in the regulation of receptor numbers at synapses.

Animals↗

PubMed2006

AMPA signalling in nascent glutamatergic synapses: there and not there!

Nascent glutamatergic synapses are thought to be equipped with only NMDA receptors and to mature in a stepwise fashion when AMPA receptors are acquired later, through specific patterns of activity. We review recent data suggesting that AMPA receptors are in fact present in the nascent synapse but in a labile state. The nascent synapse can easily switch between AMPA-signalling and AMPA-silent states in a manner not requiring activation of NMDA receptors or metabotropic glutamate receptors. NMDA receptor activation by correlated presynaptic and postsynaptic activity can switch the nascent synapse to a mature, more stable state, in which AMPA receptor signalling is modified only through conventional plasticity processes. Thus, the AMPA receptor silence of nascent glutamatergic synapses depends on the synaptic activation history rather than on the nascent state itself.

Animals↗

PubMed2005

Behavioral and neurochemical repercussions of hippocampal network activity blockade during the neonatal period.

Early destruction of the ventral hippocampus from postnatal day 7 (P7) has been shown to induce behavioral alterations in post-pubertal rats, similar to those observed in models for schizophrenia. Using a single injection of tetanus toxin into the ventral hippocampus at P1, we tested the consequences of an early neonatal activity deprivation (<P7) on behavioral and neurochemical parameters of pre- and post-pubertal rats. We found no significant differences in either behavioral or biochemical pattern, indicating that an early neonate neural activity blockade does not induce behavioral alterations in pre- and post-puberty rats.

Animals↗

PubMed2005

Resveratrol-induced apoptotic death in human U251 glioma cells.

Resveratrol (trans-3,4',5-trihydroxystilbene) is a naturally occurring polyphenolic compound highly enriched in grapes, peanuts, red wine, and a variety of food sources. Resveratrol has antiinflammatory and antioxidant properties, and also has potent anticancer properties. Human glioma U251 cells were used to understand the molecular mechanisms by which resveratrol acts as an anticancer agent, since glioma is a particularly difficult cancer to treat and eradicate. Our data show that resveratrol induces dose- and time-dependent death of U251 cells, as measured by lactate dehydrogenase release and internucleosomal DNA fragmentation assays. Resveratrol induces activation of caspase-3 and increases the cleavage of the downstream caspase substrate, poly(ADP-ribose) polymerase. Resveratrol-induced DNA fragmentation can be completely blocked by either a general caspase inhibitor (Z-VAD-FMK) or a selective caspase-3 inhibitor (Z-DEVD-FMK), but not by a selective caspase-1 inhibitor. Resveratrol induces cytochrome c release from mitochondria to the cytoplasm and activation of caspase-9. Resveratrol also increases expression of proapoptotic Bax and its translocation to the mitochondria. Resveratrol inhibits U251 proliferation, as measured by MTS assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt], and induces G0/G1 growth arrest, as determined by flow cytometry. The cyclin-dependent kinase inhibitor, olomoucine, prevents cell cycle progression and resveratrol-induced apoptosis. These results suggest that multiple signaling pathways may underlie the apoptotic death of U251 glioma induced by resveratrol, which warrants further exploration as an anticancer agent in human glioma.

Amino Acid Chloromethyl Ketones↗

PubMed2005

Roles of Ras-Erk in apoptosis of PC12 cells induced by trophic factor withdrawal or oxidative stress.

To understand the role of Ras-MAPK (mitogen-activated protein kinase) in trophic factor withdrawal- and oxidative stress-induced apoptotic cell death processes, undifferentiated rat pheochromocytoma PC12 cells and a PC12 variant cell line stably expressing the Ras dominant-negative mutant (M-M17-26) were subjected to serum withdrawal in the absence or presence of H2O2 treatment. The extent of cell death was analyzed by lactate dehydrogenase release, internucleosomal DNA fragmentation, and caspase-3 assays. Both serum withdrawal and H2O2 treatment induced apoptotic cell death in PC12 cells, and the extent of cell death was greatly enhanced in M-M17-26 cells. DNA fragmentation induced by serum withdrawal or H2O2 treatment was blocked completely by a general caspase inhibitor, Z-VAD-FMK. A selective MAPK kinase inhibitor, U0126, blocked the H2O2-induced phosphorylation of Erk1/2 (extracellular signal-regulated kinase) in PC12 cells and increased the levels of active caspase-3 in M-M17-26 under serum withdrawal or H2O2 treatment. In addition, the short-term H2O2 treatment (5-30 min) was sufficient to cause DNA fragmentation in M-M17-26 cells even though H2O2 was removed and cells were incubated in regular growth medium with complete serum for 24 h. However, similar, short-term H2O2 treatment of PC12 cells did not induce DNA fragmentation 24 h later. These results suggest that the Ras-Erk pathway is critical in mediating protection against apoptotic cell death induced by either trophic factor withdrawal or increased oxidative stress.

Animals↗

PubMed2004

Differential activity-dependent regulation of the lateral mobilities of AMPA and NMDA receptors.

The basis for differences in activity-dependent trafficking of AMPA receptors (AMPARs) and NMDA receptors (NMDARs) remains unclear. Using single-molecule tracking, we found different lateral mobilities for AMPARs and NMDARs: changes in neuronal activity modified AMPAR but not NMDAR mobility, whereas protein kinase C activation modified both. Differences in mobility were mainly detected for extrasynaptic AMPARs, suggesting that receptor diffusion between synaptic and extrasynaptic domains is involved in plasticity processes.

Animals↗

PubMed2003

Kainic acid lesion-induced nigral neuronal death.

Parkinson's disease (PD) is characterized by progressive death of dopamine (DA) neurons in the substantia nigra pars compacta. We report a rat model that exhibits progressive death of nigral neurons following unilateral injection of kainic acid in the striatum. In situ end-labeling revealed significant numbers of dying nigral neurons ipsilateral to the lesion during the first 3 weeks following injection. An indication of the gradual nature of death was that similar small numbers of cells were detected at each time point. These early morphological markers of neuronal death led to a significant reduction (20%) at 5 months of tyrosine hydroxylase-positive neurons and total number of neurons in the ipsilateral substantia nigra compared with the contralateral control. To examine the role of nigrostriatal DA metabolism in the observed nigral neuronal death, we manipulated DA metabolism during the initial 2 weeks following kainic acid lesion. Neurons in the ventral tier of the substantia nigra pars compacta were protected from death by treatment with 2,4-diamino-6-hydroxy-pyrimidine (DAHP), an inhibitor of GTP cyclohydrolase, the initial enzyme in the synthesis of the tyrosine hydroxylase co-substrate, tetrahydrobiopterin (BH(4)). Neurons in both the dorsal and ventral tier of substantia nigra pars compacta were protected from death by treatment with DAHP and L-DOPA. These experiments suggest that intrastriatal kainic acid lesion is an in vivo model of trophic support withdrawal. This experimental procedure is useful for studying mechanisms underlying protracted death of nigral DA neurons and may provide valuable mechanistic information relevant to understanding the etiology of PD.

Animals↗

PubMed2003

Early establishment of multiple release site connectivity between interneurons and pyramidal neurons in the developing hippocampus.

The strength of the synaptic transmission between two neurons critically depends on the number of release sites connecting the neurons. Here we examine the development of connectivity between gamma-aminobutyric acid (GABA)ergic interneurons and CA1 pyramidal neurons in the hippocampus. GABAergic postsynaptic currents (PSCs) were recorded in whole-cell voltage-clamped CA1 pyramidal neurons. By comparing spontaneous and miniature (action potential-independent) GABAergic PSCs, we found that multiple release site connectivity is established already at the first postnatal day and that the degree of connectivity remains unaltered into adulthood. During the same time there is a dramatic increase in the number of GABAergic synapses on each pyramidal neuron as indicated by the increase in frequency of miniature GABAergic PSCs. These results indicate that during development a given interneuron contacts an increasing number of target pyramidal neurons but with the same multiple release site connectivity. It has been shown previously that the connectivity between CA3 and CA1 pyramidal neurons is initially restricted to one release site, and develops gradually. The present result thus suggests different mechanisms to govern the maturation of excitatory and inhibitory synaptic transmissions.

Animals↗

PubMed2003

Compensatory dendritic growth of CA1 pyramidal cells following growth impairment in the neonatal period.

In the neonatal rat, inhibition of hippocampal neural activity in vivo by tetanus toxin results in a severe growth impairment of the basal dendrites of CA1 pyramidal cells. Here we tested whether this early growth impairment results in a permanent reduction of the basal dendritic tree or whether recovery processes are recruited later in development when synaptic activity has fully recovered. Quantitative analysis of dendritic parameters and spine density from reconstructed CA1 pyramidal cells showed that young adult CA1 pyramidal (postnatal day 31-34) cells that were exposed to activity deprivation in the neonatal period were almost indistinguishable from control cells of the same age. These results suggest that the early hippocampal activity controls the growth rate but is not necessary for the generation of an adult normal basal dendritic tree.

Aging↗

PubMed2002

Nitric oxide synthase inhibition during development: effect on apoptotic death of dopamine neurons.

Naturally occurring cell death via apoptosis occurs in the substantia nigra pars compacta (SNc) during rat development, culminating during the perinatal period. We previously showed that lipid peroxidation-mediated oxidative stress is not involved in this cell death process. Nitric oxide (NO) has been proposed to be critical for many developmental processes in brain and has been shown to mediate cell death in neurotoxin models of neurodegenerative disorders. Here, we reported that in vivo pre- and postnatal treatment with the non-specific NO synthase (NOS) inhibitor, L-NAME (60 mg/kg), or with the neuronal NOS inhibitor, 7-NI (30 mg/kg), dramatically decreased the NOS activity as well as the NADPH-diaphorase staining in brain. However, those treatments did not rescue dopamine neurons from developmental death, suggesting that NO is not involved in vivo in developmental death of these neurons or in the overall development of the SNc.

Animals↗

PubMed2002

Spontaneous unitary synaptic activity in CA1 pyramidal neurons during early postnatal development: constant contribution of AMPA and NMDA receptors.

Maturation of the glutamatergic synapse is thought to require the incorporation of AMPA receptors at pure NMDA synapses, also called "silent" synapses. However, the relative number of silent synapses at different developmental stages, and even the concept that silent synapses lack AMPA receptors, is actively debated. In the present work, spontaneous synaptic events were used to investigate the relative contribution of synaptic AMPA and NMDA receptor-mediated transmission in CA1 pyramidal cells during the early postnatal development. Spontaneous postsynaptic currents, mediated by AMPA and NMDA receptors, were recorded from visualized CA1 pyramidal neurons over the first postnatal week. AMPA/NMDA ratio for frequency was close to one, and, importantly, it was constant over the first postnatal week. These findings suggest that the vast majority of nascent glutamatergic synapses express both functional AMPA and NMDA receptors in the neonatal hippocampus.

Animals↗

PubMed2002

In vivo blockade of neural activity alters dendritic development of neonatal CA1 pyramidal cells.

During development, neural activity has been proposed to promote neuronal growth. During the first postnatal week, the hippocampus is characterized by an oscillating neural network activity and a rapid neuronal growth. In the present study we tested in vivo, by injecting tetanus toxin into the hippocampus of P1 rats, whether this neural activity indeed promotes growth of pyramidal cells. We have previously shown that tetanus toxin injection leads to a strong reduction in the frequency of spontaneous GABA and glutamatergic synaptic currents, and to a complete blockade of the early neural network activity during the first postnatal week. Morphology of neurobiotin-filled CA1 pyramidal cells was analyzed at the end of the first postnatal week (P6-10). In activity-reduced neurons, the total length of basal dendritic tree was three times less than control. The number, but not the length, of basal dendritic branches was affected. The growth impairment was restricted to the basal dendrites. The apical dendrite, the axons, or the soma grew normally during activity deprivation. Thus, the in vivo neural activity in the neonate hippocampus seems to promote neuronal growth by initiating novel branches.

Animals↗