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Cell Synapses (cell + synapsis)
Kinds of Cell Synapses Selected AbstractsFunctions of glutamate transporters in cerebellar Purkinje cell synapsesACTA PHYSIOLOGICA, Issue 1 2009Y. Takayasu Abstract Glutamate transporters play a critical role in the maintenance of low extracellular concentrations of glutamate, which prevents the overactivation of post-synaptic glutamate receptors. Four distinct glutamate transporters, GLAST/EAAT1, GLT-1/EAAT2, EAAC1/EAAT3 and EAAT4, are distributed in the molecular layer of the cerebellum, especially near glutamatergic synapses in Purkinje cells (PCs). This review summarizes the current knowledge about the differential roles of these transporters at excitatory synapses of PCs. Data come predominantly from electrophysiological experiments in mutant mice that are deficient in each of these transporter genes. GLAST expressed in Bergmann glia contributes to the clearing of the majority of glutamate that floods out of the synaptic cleft immediately after transmitter release from the climbing fibre (CF) and parallel fibre (PF) terminals. It is indispensable to maintain a one-to-one relationship in synaptic transmission at the CF synapses by preventing transcellular glutamate spillover. GLT-1 plays a similar but minor role in the uptake of glutamate as GLAST. Although the loss of neither GLAST nor GLT-1 affects cerebellar morphology, the deletion of both GLAST and GLT-1 genes causes the death of the mutant animal and hinders the folium formation of the cerebellum. EAAT4 removes the low concentrations of glutamate that escape from uptake by glial transporters, preventing the transmitter from spilling over into neighbouring synapses. It also regulates the activation of metabotropic glutamate receptor 1 (mGluR1) in perisynaptic regions at PF synapses, which in turn affects mGluR1-mediated events including slow EPSCs and long-term depression. No change in synaptic function is detected in mice that are deficient in EAAC1. [source] TARPs ,-2 and ,-7 are essential for AMPA receptor expression in the cerebellumEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 12 2010Maya Yamazaki Abstract The ,-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors require auxiliary subunits termed transmembrane AMPA receptor regulatory proteins (TARPs), which promote receptor trafficking to the cell surface and synapses and modulate channel pharmacology and gating. Of six TARPs, ,-2 and ,-7 are the two major TARPs expressed in the cerebellum. In the present study, we pursued their roles in synaptic expression of cerebellar AMPA receptors. In the cerebellar cortex, ,-2 and ,-7 were preferentially localized at various asymmetrical synapses. Using quantitative Western blot and immunofluorescence, we found severe reductions in GluA2 and GluA3 and mild reduction in GluA4 in ,-2-knockout (KO) cerebellum, whereas GluA1 and GluA4 were moderately reduced in ,-7-KO cerebellum. GluA2, GluA3 and GluA4 were further reduced in ,-2/,-7 double-KO (DKO) cerebellum. The large losses of GluA2 and GluA3 in ,-2-KO mice and further reductions in DKO mice were confirmed at all asymmetrical synapses examined with postembedding immunogold. Most notably, the GluA2 level in the postsynaptic density fraction, GluA2 labeling density at parallel fiber,Purkinje cell synapses, and AMPA receptor-mediated currents at climbing fiber,Purkinje cell synapses were all reduced to approximately 10% of the wild-type levels in DKO mice. On the other hand, the reduction in GluA4 in ,-7-KO granular layer reflected its loss at mossy fiber,granule cell synapses, whereas that of GluA1 and GluA4 in ,-7-KO molecular layer was caused, at least partly, by their loss in Bergmann glia. Therefore, ,-2 and ,-7 cooperatively promote synaptic expression of cerebellar AMPA receptors, and the latter also promotes glial expression. [source] Involvement of post-synaptic kainate receptors during synaptic transmission between unitary connections in rat neocortexEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 11 2003Afia B. Ali Abstract The properties of functional kainate receptor-mediated EPSCs were studied in acute slices from 19,35-day-old rats. EPSCs elicited in pyramidal and fast-spiking cells in layers 2/3 and 5 of the rat motor cortex by extracellular single shock stimulus in the presence of GYKI 53655 and D-2-amino-5-phosphopentanoic resulted in a residual current. This current was not enhanced by cyclothiazide but was blocked by 6-cyano-7-nitroquinoxalin-2,3-dione and is thought to be mediated by kainate receptors. These kainate receptor-mediated currents displayed a wide range of time courses depending on which pre-synaptic fibres were activated. With paired recordings, unitary EPSCs elicited in pyramidal cells were almost totally blocked by GYKI 53655 and D-2-amino-5-phosphopentanoic. However, when L-transpyrrolidine-2,4-dicarboxylate (PDC), a glutamate uptake blocker, was introduced in the bath, the amplitude of kainate receptor-mediated currents, which is resistant to GYKI 53655 and D-2-amino-5-phosphopentanoic, was revealed. The rise and decay time constants of the kainate receptor-mediated currents were identical to control EPSCs. PDC was not required to reveal the kainate receptor-mediated currents elicited in fast-spiking cells which also displayed similar rise and decay time constants to the control EPSCs. Excitatory input onto pyramidal and fast-spiking cells in the neocortex mediated by kainate receptors contributed between 14 and 40% of the total control unitary EPSCs which displayed identical time courses to the AMPA receptor-mediated component of the EPSCs. Post-synaptic kainate receptors at connected pyramidal cell synapses may be located extra-synaptically. [source] Impairment of eyeblink conditioning in GluR,2-mutant mice depends on the temporal overlap between conditioned and unconditioned stimuliEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2001Yasushi Kishimoto Abstract Mice lacking the glutamate receptor subunit ,2 (GluR,2) are deficient in cerebellar long-term depression (LTD) at the parallel fibre,Purkinje cell synapses. We conducted delay and trace eyeblink conditioning with these mice, using various temporal intervals between the conditioned stimulus (CS) and unconditioned stimulus (US). During trace conditioning in which a stimulus-free trace interval (TI) of 250, 100 or 50 ms intervened between the 352-ms tone CS and 100-ms US, GluR,2-mutant mice learned as successfully as wild-type mice. Even in the paradigm with TI = 0 ms, in which the end of CS and onset of US are simultaneous, there was no difference between the GluR,2-mutant and wild-type mice in their acquisition of a conditioned response. However, in the delay paradigm in which the 452-ms CS overlapped temporally with the coterminating 100-ms US, GluR,2-mutant mice exhibited severe learning impairment. The present study together with our previous work [Kishimoto, Y., Kawahara, S., Suzuki, M., Mori, H., Mishina, M. & Kirino, Y. (2001) Eur. J. Neurosci.,13, 1249,1254], indicates that cerebellar LTD-independent learning is possible in paradigms without temporal overlap between the CS and US. On the other hand, GluR,2 and cerebellar LTD are essential for learning when there is CS,US temporal overlap, suggesting that the cerebellar neural substrates underlying eyeblink conditioning may change, depending on the temporal overlap of the CS and US. [source] NMDA receptor subunits GluR,1, GluR,3 and GluR,1 are enriched at the mossy fibre,granule cell synapse in the adult mouse cerebellumEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 11 2001Kazuyuki Yamada Abstract Cerebellar N -methyl- d -aspartate (NMDA) receptors are concentrated in the granular layer and are involved in motor coordination and the induction of long-term potentiation at mossy fibre,granule cell synapses. In the present study, we used immunohistochemistry to examine the distribution of NMDA receptor subunits in the adult mouse cerebellum. We found that appropriate pepsin pretreatment of sections greatly enhanced the sensitivity and specificity of immunohistochemical detection. As a result, intense immunolabelling for GluR,1 (NR2A), GluR,3 (NR2C), and GluR,1 (NR1) all appeared in synaptic glomeruli of the granular layer. Double immunofluorescence showed that these subunits were colocalized in individual synaptic glomeruli. Within the glomerulus, NMDA receptor subunits were located between centrally-located huge mossy fibre terminals and peripherally-located tiny Golgi axon terminals. By immunoelectron microscopy, all three subunits were detected at the postsynaptic junction in granule cell dendrites, forming synapses with mossy fibre terminals. Consistent with the known functional localization, GluR,1, GluR,3, and GluR,1 are, thus, anatomically concentrated at the mossy fibre,granule cell synapse. By contrast, immunohistochemical signals were very low in Purkinje cell somata and dendrites in the molecular layer. The lack of GluR,1 immunolabelling in Purkinje cells was unexpected because the cells express GluR,1 mRNA at high levels and high levels of GluR,1 protein in the molecular layer were revealed by immunoblot. As Purkinje cells are exceptionally lacking GluR, expression, the discrepant result may provide in vivo evidence suggesting the importance of accompanying GluR, subunits in synaptic localization of GluR,1. [source] Deficient long-term synaptic depression in the rostral cerebellum correlated with impaired motor learning in phospholipase C ,4 mutant miceEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 10 2001Mariko Miyata Abstract Long-term depression (LTD) at parallel fibre,Purkinje cell synapse of the cerebellum is thought to be a cellular substrate for motor learning. LTD requires activation of metabotropic glutamate receptor subtype 1 (mGluR1) and its downstream signalling pathways, which invariably involves phospholipase C,s (PLC,s). PLC,s consist of four isoforms (PLC,1,4) among which PLC,4 is the major isoform in most Purkinje cells in the rostral cerebellum (lobule 1 to the rostral half of lobule 6). We studied mutant mice deficient in PLC,4, and found that LTD was deficient in the rostral but not in the caudal cerebellum of the mutant. Basic properties of parallel fibre,Purkinje cell synapses and voltage-gated Ca2+ channel currents appeared normal. The mGluR1-mediated Ca2+ release induced by repetitive parallel fibre stimulation was absent in the rostral cerebellum of the mutant, suggesting that their LTD lesion was due to the defect in the mGluR1-mediated signalling in Purkinje cells. Importantly, the eyeblink conditioning, a simple form of discrete motor learning, was severely impaired in PLC,4 mutant mice. Wild-type mice developed the conditioned eyeblink response, when pairs of the conditioned stimulus (tone) and the unconditioned stimulus (periorbital shock) were repeatedly applied. In contrast, PLC,4 mutant mice could not learn the association between the conditioned and unconditioned stimuli, although their behavioural responses to the tone or to the periorbital shock appeared normal. These results strongly suggest that PLC,4 is essential for LTD in the rostral cerebellum, which may be required for the acuisition of the conditioned eyeblink response. [source] Differential sensitivity to Zolpidem of IPSPs activated by morphologically identified CA1 interneurons in slices of rat hippocampusEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 2 2000Alex M. Thomson Abstract Hippocampal pyramidal cells express several ,-subunits, which determine the affinity of GABAA (,-aminobutyric acid) receptors for benzodiazepine site ligands. This study asked whether inhibitory postsynaptic potentials (IPSPs) elicited by specific interneuronal subclasses were differentially sensitive to the ,1-preferring agonist Zolpidem, i.e. whether different receptors mediate different inhibitory connections. Paired intracellular recordings in which the presynaptic cell was an interneuron and the postsynaptic cell a CA1 pyramid were performed in slices of adult rat hippocampus. Resultant IPSPs were challenged with Zolpidem, cells filled with biocytin and identified morphologically. IPSPs elicited by fast spiking (FS) basket cells (n = 9) were enhanced more than IPSPs elicited by regular spiking (RS) basket cells (n = 10). At FS basket cell synapses the efficacy of Zolpidem was equivalent to that of Diazepam, while RS basket cell IPSPs are enhanced 50% less by Zolpidem than by Diazepam. Thus, while ,1 subunits may dominate at synapses supplied by FS basket cells, RS basket cell synapses also involve ,2/3 subunits. Two bistratified cell IPSPs tested with Zolpidem did not increase in amplitude, despite powerful enhancements of bistratified cell IPSPs by Diazepam, consistent with previous indications that these synapses utilize ,5-containing receptors. Enhancements of basket cell IPSPs by Zolpidem and Diazepam were bi- or triphasic with steep amplitude increases separated by plateaux, occurring 10,15, 25,30 and 45,55 min after adding the drug to the bath. The entire enhancement was, however, blocked by the antagonist Flumazenil (n = 7). Flumazenil, either alone (n = 3), or after Zolpidem, reduced IPSP amplitude to ,,90% of control, suggesting that ,4-containing receptors were not involved. [source] Calcium signaling in specialized glial cells,GLIA, Issue 7 2006Monica R. Metea Abstract This article reviews calcium signaling in three specialized types of glial cells: Müller cells of the retina, Bergmann glial cells of the cerebellum, and radial glial cells of the developing cortex. Müller cells generate spontaneous and neuronal activity-evoked increases in Ca2+. Neuron to Müller cell signaling is mediated by neuronal release of ATP and activation of glial P2Y receptors. Müller cells, in turn, modulate neuronal excitability and mediate vasomotor responses. Bergmann glial cells also generate spontaneous and activity-evoked Ca2+ increases. Neuron to Bergmann glia signaling is mediated by neuronal release of nitric oxide, noradrenaline, and glutamate. In Bergmann glia, Ca2+ increases control the structural and functional interactions between these cells and Purkinje cell synapses. In the ventricular zone of the developing cortex, radial glial cells generate spontaneous Ca2+ increases that propagate as Ca2+ waves through clusters of neighboring glial cells. These Ca2+ increases control cell proliferation and neurogenesis. © 2006 Wiley-Liss, Inc. [source] IL-4 attenuates the neuroinflammation induced by amyloid-, in vivo and in vitroJOURNAL OF NEUROCHEMISTRY, Issue 3 2007Anthony Lyons Abstract It has been shown that A, inhibits long-term potentiation (LTP) in the rat hippocampus and this is accompanied by an increase in hippocampal concentration of IL-1,. A, also increases microglial activation, which is the likely cell source of IL-1,. Because IL-4 attenuates the effects of IL-1, in hippocampus, and microglial activation is inhibited by minocycline, we assessed the ability of both IL-4 and minocycline to modulate the effects of A, on LTP and IL-1, concentration. Following treatment with A,, IL-4 or minocycline, rats were assessed for their ability to sustain LTP in perforant path-granule cell synapses. We report that the A,-induced inhibition of LTP was associated with increases in expression of MHCII, JNK phosphorylation and IL-1, concentration, and that these changes were attenuated by treatment of rats with IL-4 and minocycline. We also report that A,-induced increases in expression of MHCII and IL-1, were similarly attenuated by IL-4 and minocycline in glial cultures prepared from neonatal rats. These data suggest that glial cell activation and the consequent increase in IL-1, concentration mediate the inhibitory effect of A, on LTP and indicate that IL-4, by down-regulating glial cell activation, antagonizes the effects of A,. [source] Synaptic localization of P2X7 receptors in the rat retinaTHE JOURNAL OF COMPARATIVE NEUROLOGY, Issue 1 2004Theresa Puthussery Abstract The distribution of P2X7 receptor (P2X7R) subunits was studied in the rat retina using a subunit-specific antiserum. Punctate immunofluorescence was observed in the inner and outer plexiform layers. Double labeling of P2X7 and the horizontal cell marker, calbindin, revealed extensive colocalization in the outer plexiform layer (OPL). Significant colocalization of P2X7R and kinesin, a marker of photoreceptor ribbons, was also observed, indicating that this receptor may be expressed at photoreceptor terminals. Furthermore, another band of P2X7R puncta was identified below the level of the photoreceptor terminals, adjacent to the inner nuclear layer (INL). This band of P2X7R puncta colocalized with the active-zone protein, bassoon, suggesting that "synapse-like" structures exist outside photoreceptor terminals. Preembedding immunoelectron microscopy demonstrated P2X7R labeling of photoreceptor terminals adjacent to ribbons. In addition, some horizontal cell dendrites and putative "desmosome-like" junctions below cone pedicles were labeled. In the inner plexiform layer (IPL), P2X7R puncta were observed surrounding terminals immunoreactive for protein kinase C-,, a marker of rod bipolar cells. Double labeling with bassoon in the IPL revealed extensive colocalization, indicating that P2X7R is likely to be found at conventional cell synapses. This finding was confirmed at the ultrastructural level: only processes presynaptic to rod bipolar cells were found to be labeled for the P2X7R, as well as other conventional synapses. These findings suggest that purines play a significant role in neurotransmission within the retina, and may modulate both photoreceptor and rod bipolar cell responses. J. Comp. Neurol. 472:13,23, 2004. © 2004 Wiley-Liss, Inc. [source] Adenosine signalling at immature parallel fibre,Purkinje cell synapses in rat cerebellumTHE JOURNAL OF PHYSIOLOGY, Issue 18 2009Alison Atterbury The purine adenosine is an extracellular signalling molecule involved in a large number of physiological and pathological conditions throughout the mammalian brain. However little is known about how adenosine release and its subsequent clearance change during brain development. We have combined electrophysiology and microelectrode biosensor measurements to investigate the properties of adenosine signalling at early stages of cerebellar development, when parallel fibre,Purkinje cell synapses have recently been formed (postnatal days 9,12). At this stage of development, we could detect little or no inhibitory A1 receptor tone in basal conditions and during trains of stimuli. Addition of pharmacological agents, to inhibit adenosine clearance, had only minor effects on synaptic transmission suggesting that under basal conditions, the concentration of adenosine moving in and out of the extracellular space is small. Active adenosine release was stimulated with hypoxia and trains of electrical stimuli. Although hypoxia released significant concentrations of adenosine, the release was delayed and slow. No adenosine release could be detected following electrical stimulation in the molecular layer. In conclusion, at this stage of development, although adenosine receptors and the mechanisms of adenosine clearance are present there is very little adenosine release. [source] Sustained granule cell activity disinhibits juvenile mouse cerebellar stellate cells through presynaptic mechanismsTHE JOURNAL OF PHYSIOLOGY, Issue 2 2008Simone Astori GABA release from cerebellar molecular layer interneurons can be modulated by presynaptic glutamate and/or GABAB receptors upon perfusing the respective agonists. However, it is unclear how release and potential spillover of endogenous transmitter lead to activation of presynaptic receptors. High frequency firing of granule cells, as observed in vivo upon sensory stimulation, could lead to glutamate and/or GABA spillover. Here, we established sustained glutamatergic activity in the granule cell layer of acute mouse cerebellar slices and performed 190 paired recordings from connected stellate cells. Train stimulation at 50 Hz reduced by about 30% the peak amplitude of IPSCs evoked by brief depolarization of the presynaptic cell in 2-week-old mice. A presynaptic mechanism was indicated by changes in failure rate, paired-pulse ratio and coefficient of variation of evoked IPSCs. Furthermore, two-photon Ca2+ imaging in identified Ca2+ hot spots of stellate cell axons confirmed reduced presynaptic Ca2+ influx after train stimulation within the granular layer. Pharmacological experiments indicated that glutamate released from parallel fibres activated AMPARs in stellate cells, evoking GABA release from surrounding cells. Consequential GABA spillover activated presynaptic GABABRs, which reduced the amplitude of eIPSCs. Two-thirds of the total disinhibitory effect were mediated by GABABRs, one-third being attributable to presynaptic AMPARs. This estimation was confirmed by the observation that bath applied baclofen induced a more pronounced reduction of evoked IPSCs than kainate. Granule cell-mediated disinhibition persisted at near-physiological temperature but was strongly diminished in 3-week-old mice. At this age, GABA release probability was not reduced and presynaptic GABABRs were still detectable, but GABA uptake appeared to be advanced, attenuating GABA spillover. Thus, sustained granule cell activity modulates stellate cell-to-stellate cell synapses, involving transmitter spillover during a developmentally restricted period. [source] Auto-inhibition of rat parallel fibre,Purkinje cell synapses by activity-dependent adenosine releaseTHE JOURNAL OF PHYSIOLOGY, Issue 2 2007Mark J. Wall Adenosine is an important signalling molecule involved in a large number of physiological functions. In the brain these processes are as diverse as sleep, memory, locomotion and neuroprotection during episodes of ischaemia and hypoxia. Although the actions of adenosine, through cell surface G-protein-coupled receptors, are well characterized, in many cases the sources of adenosine and mechanisms of release have not been defined. Here we demonstrate the activity-dependent release of adenosine in the cerebellum using a combination of electrophysiology and biosensors. Short trains of electrical stimuli delivered to the molecular layer in vitro, release adenosine via a process that is both TTX and Ca2+ sensitive. As ATP release cannot be detected, adenosine must either be released directly or rapidly produced by highly localized and efficient extracellular ATP breakdown. Since adenosine release can be modulated by receptors that act on parallel fibre,Purkinje cell synapses, we suggest that the parallel fibres release adenosine. This activity-dependent adenosine release exerts feedback inhibition of parallel fibre,Purkinje cell transmission. Spike-mediated adenosine release from parallel fibres will thus powerfully regulate cerebellar circuit output. [source] NO signalling decodes frequency of neuronal activity and generates synapse-specific plasticity in mouse cerebellumTHE JOURNAL OF PHYSIOLOGY, Issue 3 2005Shigeyuki Namiki Nitric oxide (NO) is an intercellular messenger regulating neuronal functions. To visualize NO signalling in the brain, we generated a novel fluorescent NO indicator, which consists of the heme-binding region (HBR) of soluble guanylyl cyclase and the green fluorescent protein. The indicator (HBR,GFP) was expressed in the Purkinje cells of the mouse cerebellum and we imaged NO signals in acute cerebellar slices upon parallel fibre (PF) activation with a train of burst stimulations (BS, each BS consisting of five pulses at 50 Hz). Our results showed that the intensity of synaptic NO signal decays steeply with the distance from the synaptic input near PF,Purkinje cell synapses and generates synapse-specific long-term potentiation (LTP). Furthermore, the NO release level has a bell-shaped dependence on the frequency of PF activity. At an optimal frequency (1 Hz), but not at a low frequency (0.25 Hz) of a train of 60 BS, NO release as well as LTP was induced. However, both NO release and LTP were significantly reduced at higher frequencies (2,4 Hz) of BS train due to cannabinoid receptor-mediated retrograde inhibition of NO generation at the PF terminals. These results suggest that synaptic NO signalling decodes the frequency of neuronal activity to mediate synaptic plasticity at the PF,Purkinje cell synapse. [source] | ||||||||||