1,721,006 research outputs found
Postsynaptic currents in deep cerebellar nuclei
No full textPostsynaptic currents were studied by whole cell recordings in visually identified large neurons of the deep cerebellar nuclei (DCN) in slices of 4- to 11-day-old mice, Spontaneous postsynaptic currents were abolished by the GABAA receptor antagonist bicuculline and had a single-exponential decay with a mean time constant of 13.6 ± 3.2 (SD) ms. Excitatory postsynaptic currents (EPSCs) were evoked in 48/56 neurons recorded. The addition of AMPA and N-methyl-D-aspartate (NMDA) receptor antagonists together completely abolished all synaptic responses. In 1 mM [Mg2+]o and at a holding potential of -60 mV, the peak amplitude of the NMDA component of the EPSC (NMDA-EPSC) was 83.2 ± 21.2% of the AMPA component (AMPA-EPSC). This indicates that in DCN neurons, at a physiological [Mg2+]o and at the resting membrane potential, NMDA receptors contribute to the synaptic signal. AMPA-EPSCs had a linear current-voltage relationship with a reversal potential of +2.3 ± 0.4 mV and a single-exponential decay with a voltage-dependent time constant that at -60 mV was 7.1 ± 3.3 ms. In 10 μM glycine and 1 mM [Mg2+]o, the I-V relationship of NMDA-EPSCs had a reversal potential of -0.5 ± 3.3 mV and a maximal inward current at -33.4 ± 5.8 mV. The apparent dissociation constant (KD) of Mg2+ for the NMDA receptor-channel at -60 mV, measured by varying [Mg2+]o, was 135.5 ± 55.3 μM, and when measured by fitting the I-V curves with a theoretical function, it was 169.9 ± 119.5 μM. Thus in the DCN, NMDA receptors have a sensitivity to Mg2+ that corresponds to subunits that are weakly blocked by this ion (ε3 and ε4) of which the DCN express ε4. NMDA-EPSCs had a double-exponential decay with voltage-dependent time constants that at -60 mV were 20.2 ± 8.9 and 136.4 ± 62.8 ms. At positive voltages, the time constants were slower and their contributions were about equal, while in the negative slope conductance region of the I-V curve, the faster time constant became predominant, conferring faster kinetics to the EPSC. The weak sensitivity to Mg2+ of NMDA receptors, together with a relatively fast kinetics, provide DCN neurons with strong excitatory inputs in which fast dynamic signals are relatively well preserved
Spontaneous saccades and gaze holding ability in the pigmented rat: I. Effects of inferior olive lesion.
No full textWe have studied the effects of lesion of the inferior olive on the spontaneous eye movements performed both in the light and dark in head restrained pigmented rats. The inferior olive lesion was made at least 1 month before study with 3-acetylpyridine and eye movements were recorded through a phase detection search coil apparatus. Following lesion, the spontaneous saccades performed in the dark present a postsaccadic drift which is made up of two components characterized by their different time courses, the first one being fast and the second one slow. The latter component is due to the leakage of the neural integrator and the former is mainly the consequence of a mismatch between the phasic and the tonic component of the ocular movement. In the light only the first component is present and then the eye maintains a steady position. After the lesion the saccades in the dark present a time constant of the slow component of the postsaccadic drift which is significantly reduced to approximately 600 - 900 ms from a value of 1600 - 4000 ms of the intact rats. This means that the integrity of the inferior olive is necessary to keep the time constant of the neural integrator within the physiological range. In the light, the amplitude of the postsaccadic drift depends on two factors. First, there is a mismatch between the phasic and the tonic components of the ocular movement, which are due to the pulse and the step of innervation of the extraocular muscles respectively. Different types of analysis have shown that the gain of the pulse to step transformation is about 0.77 at all saccadic amplitudes and eccentricities. Second, there is an increased leakiness of the neural integrator. Such a contribution increases linearly as a function of the eccentricity with a slope of 0.21. The main sequence of the saccades is not appreciably affected by the olivary lesion. Thus, the consequence of the inferior olive lesion may be interpreted as a general disruption of the integration process which, in physiological conditions, generates a proper and sustained oculomotor signal. More generally, it may be viewed as a loss of coordination between phasic and tonic motor commands
Elevation of intradendritic sodium concentration mediated by synaptic activation of metabotropic glutamate receptors in cerebellar Purkinje cells
No full textCerebellar Purkinje cells express both ionotropic glutamate receptors and metabotropic glutamate receptors. Brief tetanic stimulation of parallel fibers in rat and mouse cerebellar slices evokes a slow excitatory postsynaptic current in Purkinje cells that is mediated by the mGluR1 subtype of metabotropic glutamate receptors. The effector system underlying this mGluR1 EPSC has not yet been identified. In the present study, we recorded the mGluR1 EPSC using the whole-cell patch-clamp technique in combination with microfluorometric recordings of the intracellular sodium concentration ([Na+](i)) by means of the fluorescent sodium indicator SBFI. The mGluR1 EPSC was induced by local parallel fibre stimulation in the presence of the ionotropic glutamate receptor antagonists NBQX and D-APV and the GABA(A) receptor antagonists bicuculline or picrotoxin, The mGluR1 EPSC was associated with an increase in [Na+](i) that was restricted to a specific portion of the dendritic tree. The mGluR1 EPSC as well as the increase in [Na+](i) were inhibited by the mGluR antagonist S-MCPG. In the presence of NBQX, D-APV, pictrotoxin and TTX, bath application of the selective mGluR agonist 3,5-DHPG induced an elevation in [Na+](i) which extended over the whole dendritic field of the Purkinje cell. This finding demonstrates that the mGluR1-mediated postsynaptic current leads to a significant influx of sodium into the dendritic cytoplasm of Purkinje cells and thereby provides a novel intracellular signalling mechanism that might be involved in mGluR1-dependent synaptic plasticity at this synapse
Postsynaptic current mediated by metabotropic glutamate receptors in cerebellar Purkinje cells.
No full textIn rat cerebellar slices, repetitive parallel fiber stimulation evokes an inward, postsynaptic current in Purkinje cells with a fast component mediated by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptors and a slower component mediated by metabotropic glutamate receptors (mGluR). The mGluR-mediated excitatory postsynaptic current (mGluR-EPSC) is evoked selectively by parallel fiber stimulation; climbing fiber stimulation is ineffective. The mGluR-EPSC is elicited most effectively with increasing frequencies of parallel fiber stimulation, from a threshold of 10 Hz to a maximum response at similar to 100 Hz. The amplitude of the mGluR-EPSc is a linear function of the number of stimulus pulses without any apparent saturation, even with >10 pulses. Thus mGluRs at the parallel fiber-Purkinje cell synapse can function as linear detectors of the number of spikes in a burst of activity in parallel fibers. The mGluR-EPSC is present from postnatal day 15 and persists into adulthood. It is inhibited by the generic mGluR antagonist (RS)-a-methyl-4-carboxyphenylglycine and by the group I mGluR antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid at a concentration selective for mGluR1. Although the intracellular transduction pathway involves a G protein, the putative mediators of mGluR1 (phospholipase C and protein kinase C) are not directly involved, indicating that the mGluR-EPSC studied here is mediated by a different and still unidentified second-messenger pathway. Heparin, a nonselective antagonist of inositol-trisphosphate (IP3) receptors, has no significant effect on the mGluR-EPSC, suggesting that also IP3 might be not required for the response. buffering intracellular Ca2+ with a high concentration of bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid partially inhibits the mGluR-EPSC, indicating that Ca2+ is not directly responsible for the response but that resting Ca2+ levels exert a tonic potentiating effect on the mGluR-EPSC
Antagonist action of imidazobenzodiazepine Ro 15-4513 on ethanol-induced alterations of saccadic eye movements in the pigmented rat
No full textFollowing an intraperitoneal injection of ethanol (1 g/kg), spontaneous saccades performed by pigmented rats showed a backward post-saccadic drift which was still present 45 min after the administration. When 5 mg/kg of Ro 15-4513 were injected, 15 min after ethanol, there was an immediate remarkable reduction in post-saccadic drift amplitude, with a full recovery in about 30 min after Ro 15-4513 administration. This drug is thus able to reverse the effect of ethanol and to restore effectively a mechanism of motor integration
GIRK1-Mediated Inwardly Rectifying Potassium Current Is a Candidate Mechanism Behind Purkinje Cell Excitability, Plasticity, and Neuromodulation
No full textG-protein-coupled inwardly rectifying potassium (GIRK) channels contribute to the resting membrane potential of many neurons and play an important role in controlling neuronal excitability. Although previous studies have revealed a high expression of GIRK subunits in the cerebellum, their functional role has never been clearly described. Using patch-clamp recordings in mice cerebellar slices, we examined the properties of the GIRK currents in Purkinje cells (PCs) and investigated the effects of a selective agonist of GIRK1-containing channels, ML297 (ML), on PC firing and synaptic plasticity. We demonstrated that GIRK channel activation decreases the PC excitability by inhibiting both sodium and calcium spikes and, in addition, modulates the complex spike response evoked by climbing fiber stimulation. Our results indicate that GIRK channels have also a marked effect on synaptic plasticity of the parallel fiber-PC synapse, as the application of ML297 increased the expression of LTP while preventing LTD. We, therefore, propose that the recruitment of GIRK channels represents a crucial mechanism by which neuromodulators can control synaptic strength and membrane conductance for proper refinement of the neural network involved in memory storage and higher cognitive functions
Postsynaptic current mediated by metabotropic glutamate receptors in cerebellar Purkinje cells
No full textIn rat cerebellar slices, repetitive parallel fiber stimulation evokes an inward, postsynaptic current in Purkinje cells with a fast component mediated by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptors and a slower component mediated by metabotropic glutamate receptors (mGluR). The mGluR-mediated excitatory postsynaptic current (mGluR-EPSC) is evoked selectively by parallel fiber stimulation; climbing fiber stimulation is ineffective. The mGluR-EPSC is elicited most effectively with increasing frequencies of parallel fiber stimulation, from a threshold of 10 Hz to a maximum response at ~ 100 Hz. The amplitude of the mGluR-EPSC is a linear function of the number of stimulus pulses without any apparent saturation, even with > 10 pulses. Thus mGluRs at the parallel fiber-Purkinje cell synapse can function as linear detectors of the number of spikes in a burst of activity in parallel fibers. The mGluR-EPSC is present from postnatal day 15 and persists into adulthood. It is inhibited by the generic mGluR antagonist (RS)-a-methyl-4-carboxyphenylglycine and by the group I mGluR antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid at a concentration selective for mGluR1. Although the intracellular transduction pathway involves a G protein, the putative mediators of mGluR1 (phospholipase C and protein kinase C) are not directly involved, indicating that the mGluR- EPSC studied here is mediated by a different and still unidentified second- messenger pathway. Heparin, a nonselective antagonist of inositol- trisphosphate (IP3) receptors, has no significant effect on the mGluR-EPSC, suggesting that also IP3 might be not required for the response. Buffering intracellular Ca2+ with a high concentration of bis-(o-aminophenoxy)- N,N,N',N'-tetraacetic acid partially inhibits the mGluR-EPSC, indicating that Ca2+ is not directly responsible for the response but that resting Ca2+ levels exert a tonic potentiating effect on the mGluR-EPSC
Saccadic eye movements and gaze in the head-restrained pigmented rat.
No full textSpontaneous saccadic eye movements were recorded in seven head-restrained pigmented rats by means of a phase detection search coil system, both in the light and in the dark. In an illuminated environment, all the rats made numerous spontaneous saccades with an average amplitude of 13.2 deg (+/- 2.2 SD) and a maximal amplitude of 35 deg. In the dark, mean saccadic amplitude was significantly reduced to 9.2 deg (+/- 2.0 SD). Saccadic peak velocity increased linearly as a function of saccadic size, with no saturation at high amplitude values. In the light, peak velocity increase was 32.7 deg/s/deg (+/- 3.5 SD). This value is higher than that described in many other species including man and is similar to that of the monkey. Also saccadic duration increased linearly as a function of size at a rate of 1 ms/deg, which is closer to that of monkey than to that of other species including man. Both peak velocity and duration were not significantly different in the dark from those measured in the light. In the light, following a saccadic gaze shift, the rats were able to maintain a steady eye position for long periods, also at large orbital eccentricities. In the dark, on the contrary, the eye presented a drift towards the central position in the orbit. Such a drift had an exponential-like time course with a time constant of 1567 ms (+/- 829 SD), a value which is much shorter than that of cat and primates. This indicates that in the absence of a visual input, the rat has a poor gaze holding ability compared to other species
Effects of ethanol and imidazobenzodiazepine Ro 15-4513 on spontaneous saccades of the pigmented rat
No full textThe present study was aimed at investigating the alterations of the spontaneous saccadic eye movements of pigmented rats following ethanol administration. In addition we have studied the efficacy of the imidazobenzodiazepine Ro 15-4513 in reversing the effects of alcohol on saccades. The horizontal component of spontaneous eye movements was recorded by means of the magnetic field search coil technique on 11 head-restrained, pigmented rats. After the intraperitoneal injection of ethanol (1 g/kg) spontaneous saccades showed: i) a backward post-saccadic drift, with an exponential-like time course (time constant 100-150 ms); ii) a remarkable reduction of mean saccadic amplitude, up to 37% of control; iii) a significant decrease of peak velocity, which was reduced to about 80% of control. All these effects appeared and developed within a few minutes after the administration and were still present one hour later. When Ro 15-4513 (5 mg/kg) was injected i.p., 15 min after ethanol, the post-saccadic drift amplitude was immediately reduced and the drift was completely abolished within about 30 min. Mean saccadic amplitude returned to control values within a few minutes and was then steadily maintained for the following period examined (30 min). On the contrary, peak velocity showed only a slight tendency to recover which never was significant. When the same dose of Ro 15-4513 was injected alone there was no post-saccadic drift. However, mean saccadic amplitude increased, almost immediately, up to 160% of control. Its value showed a slight constant decrease in the following 30 min. Peak velocity was only slightly increased (up to 106% of control), but never was significantly different from control. Our results show that ethanol induces a remarkable impairment in the performance of spontaneous saccades. The imidazobenzodiazepine Ro 15-4513 is able to reverse completely only some of the alcohol-induced alterations, i.e. the post-saccadic drift and the reduction of saccadic amplitude, while it fails to counter efficiently the reduction of peak velocity. Ro 15-4513 exerts an intrinsic action, which is opposite to that of ethanol, on some of the saccadic parameters we have examined
Spontaneous saccades and gaze holding ability in the pigmented rat: II. Effects of localized cerebellar lesions.
No full textWe have studied the effects of the ablation of the cerebellar vermal area corresponding to lobules VI - VIII and of the flocculus - paraflocculus of both sides on the spontaneous eye movements performed in the light and in the dark in head-restrained pigmented rats. These effects have been compared with those already described for the inferior olive lesion. The cerebellar lesions were performed 1 week to 6 months in advance. Eye movements were recorded through a phase detection search coil apparatus. Following vermal topectomy, the main characteristics of the spontaneous saccades are unmodified. Following the ablation of the flocculus - paraflocculus there is no change in the saccadic main sequence. However, the spontaneous saccades in the dark present a postsaccadic drift made up of two components with different time courses, the first one being fast and the second one slow. The former is due in part to a mismatch between the phasic (the pulse) and the tonic (the step) components of the eye movements; the latter to the leakage of the neural integrator. In light only the first component is present and the eye maintains a steady position. The time constant of the neural integrator is considerably reduced to approximately 600 - 900 ms from a value of approximately 1600 - 4000 ms in the intact rats. The amplitude of the postsaccadic drift in the light depends on both the mismatch between the pulse and the step of innervation of the extraocular muscles and the increased leakiness of the neural integrator. The gain of the pulse to step transformation is reduced to approximately 0.79 at all saccadic amplitudes and eccentricities and such a reduction is due to a decreased step amplitude, while the pulse amplitude remains unchanged. The contribution of the leakage of the neural integrator to the postsaccadic drift in the light is a function of the eccentricity with a slope of 0.23. The deficits described after flocculus - paraflocculus ablation are also very similar to those described following inferior olive lesion from a quantitative point of view. The possible mechanisms of the visually activated olivocerebellar system in the control of saccadic performance and in maintaining its calibration are discussed
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