Simulated synaptic conductance in PC to DCN synapse caused by spontaneous PC spiking.

<p>(A) A representative example of the simulated synaptic conductance (G_syn) induced by PC (black) of AnR (upper panel), AnM (middle panel) and AwM (lower panel), and by corresponding realizations of an inhomogeneous Poisson process (green). Rasters: spikes belonging to patterns (black and green dotted lines: start of patterns, black and green solid lines: following spikes in patterns, blue lines: singles), numbers: number of all spikes in the 500 ms window. (B) Distribution of G_syn values for PCs (black) compared to Poisson processes (green). Bin = 0.2 nS. Red bar: bins where PCs contained significantly more G_syn values. p<0.05.</p

Regular patterns in cerebellar Purkinje cell simple spike trains.

<p>(A) Raster plot of PC SS in an anesthetized rat (AnR). (B) CV₂ distributions of SS trains recorded from anesthetized mice (AnM, left), awake mice (AwM, middle, blue: neurons in cerebral motor cortex), and mean of 92 CV₂ distributions (Pooled, right) which were significantly different from those of inhomogeneous Poisson processes with similarly modulated firing rates (p<0.05, χ² test; *: p<0.001, χ² goodness of fit residual test; red line: CV₂ = 0.2). Insets and right panel: mean±s.e.m. (black: PC, green: inhomogeneous Poisson process) (C) Extracting regular spiking patterns by setting CV₂ threshold at 0.2 (white dotted lines). White dashes: CV₂ values calculated from the two surrounding ISIs, red: first ISI of regular patterns, pink: successive ISIs in regular patterns, dark blue: ISIs not belonging to a regular pattern).</p

Regular patterns and singles related to the membrane potential (MP).

<p>Dendritic patch-clamp recording of PC in anesthetized rat (data from Loewenstein et al. 2005). Voltage trace: large spikes are complex spikes, small ones are simple spikes. Dotted black line: threshold to define up and down-states (MP = −55 mV). Raster plot at top: simple spikes were sorted as either pattern spikes (dotted red lines: start of patterns, solid red lines: following spikes in each pattern) or single spikes (blue lines). All patterns were during up-state, but singles occurred both during up (filled circles) and down (open circles) states.</p

A representative example of regular patterns in tactile stimulus evoked PC SS responses.

<p>(A) Peri-event raster plot of patterns (red) and singles (blue) during tactile stimulation in AnR. (B) Mean rate (± s.e.m.) of overall spikes (black), realization of Poisson process (green), pattern spikes (red) and singles (blue). Bin = 20 ms. (C) Simulated G_syn for the trial indicated by arrow in (A) (bin = 1 ms). (D) CV (SD/Mean) of simulated G_syn (*: p<0.001, Student t test, bin 20 ms). Black dotted line: stimulation time. (E) Mean firing rate (upper panel) and percent ISIs belonging to regular patterns (lower panel) in 200 ms before and after stimulation (upper panel) of simulated spike trains from inhomogeneous Poisson process (green) and from recorded PCs (black). *: p<0.005, Wilcoxon signed ranks test. (F) Pattern mean ISI distribution before (dotted line) and after (solid line) tactile stimulation. Inset: Pattern size distribution before (open) and after (filled) stimulation.</p

Characteristics of regular spike patterns.

<p>(A) ISI distribution of overall ISIs (black), patterns (red) and singles (blue) from a representative sample PC spike train of AnR (left), AnM (middle) and AwM (right). Insets: magnified plot of indicated area (lower) and 90 P (90 percentile, upper) of each population, *: p<0.01, Student t test. (B) The relation between pattern mean ISI and pattern size in AnR (left, cyan), AnM (middle, magenta) and AwM (right, yellow). Insets: maximum pattern mean ISI (90 percentile) of different pattern sizes. *: p<0.001, Wilcoxon signed rank test. (C) Percentage ISIs belonging to patterns (upper, *: p<0.001, Student t test), Average maximum pattern size (middle, *: p<0.001, Student t test), and Pattern size distribution (lower, p<0.05, χ² test). Cyan: AnR, magenta: AnM, yellow: AwM.</p

Effect of CV₂ threshold on patterns.

<p>(A) Mean (± s.e.m.) of normalized number of patterns in spike trains classified with different values of the CV₂ threshold, ranging from 0 to 0.5, in 92 PCs (filled circles) and in simulated spike trains from Poisson processes with similar firing rate profiles as in the PCs (open circles). Arrow: maximum number of patterns, *: range where there was no statistical difference (p>0.05). Inset: same distribution but for all possible thresholds. (B) Raster plots with indication of the spike timings belonging to patterns (red dotted lines: start of patterns, red solid lines: following spikes in each pattern) and singles (blue). Black dots: difference in classified patterns when threshold was 0.2 (upper trace) and 0.24 (lower trace).</p

Coincident patterns in nearby PC pairs in AnR.

<p>(A) Eight cross-correlograms of timings of spikes belonging to regular patterns extracted from recordings of nearby PC pairs, with each pair colored differently. Insets: cross-correlograms of the shuffled spike trains of two pairs (black) superimposed on original cross-correlogram of patterns (blue and gray: pairs showing strongest and weakest synchronization respectively). (B) The relation of pattern mean ISIs in 4 pairs in which pattern starts coincided significantly (inset: cross-correlograms of the first spikes of regular patterns in the 4 pairs). Red dotted line: diagonal.</p

α7-nAChR activation at the mossy fibre-granule cell synapse.

<p>In A, B, and C, patch-clamp recordings performed from granule cells voltage-clamped at –70 mV in cerebellar slices. Drugs were applied for 100 seconds (black bar). (<b>A</b>) EPSC amplitude changes caused by application of nicotinic agents. (<i>Left)</i> Average traces of 10 contiguous EPSCs taken from a representative experiment (1 µM nicotine). <i>(Right)</i> Time course of EPSC amplitude changes (mean ± SEM) during application of 1 µM nicotine (n = 56), 10 mM choline (n = 5), 1 µM nicotine +10 nM MLA (n = 4), 50 nM epibatidine (n = 4), 10 nM MLA (n = 5), 1µM DHβE (n = 7). (<b>B</b>) EPSC amplitude changes in C57/BL6 (n = 4) and α7-nAChR KO mice (n = 4) during application of 1 µM nicotine. Same panel layout as in A. (<b>C</b>) The effect of postsynaptic calcium buffering (0.1 mM or 10 mM intracellular BAPTA) on the action of 1 µM nicotine (black bar). <i>(Left)</i> Average traces from 10 contiguous AMPA-EPSCs and NMDA-EPSCs (isolated with 10 µM NBQX in Mg²⁺-free medium) in representative recordings. <i>(Right)</i> Ensemble effects on AMPA-EPSC PPR and NMDA-EPSC amplitude (mean ± SEM). (<b>D</b>) Immunolabelling for α7-nAChR subunit in electron micrographs at the mossy fibre-granule cell synapse. An immunopositive pre-terminal mossy fibre (mf, bordered by arrows) opens up in a large bouton, which surrounds granule cell dendrites (<i>d</i>) (scale bar 0.5 µm). The inset shows a granule cell dendrite (<i>d</i>), contacted by a mossy fibre bouton (<i>b</i>), bearing an immunopositive post-synaptic specialisation (arrow). Scale bar 0.8 µm.</p

α7-nAChR stimulation facilitates LTP <i>in vivo</i>.

<p>LFP recordings were performed from the granular layer of the Crus-IIa of the right cerebellar hemisphere in response to air-puff stimulation of the homolateral whisker pad. A TSS air-puff induction pattern was delivered at the end of a 5 min microperfusion of drugs in different combinations: TSS+Krebs’ solution (n = 7), just 50 µM nicotine (n = 6), TSS +50 µM nicotine (n = 6), TSS +100 mM choline (n = 5), TSS +50 µM nicotine +0.5 µM MLA (n = 5). <i>Left</i>, LFPs recorded before and after TSS+nicotine (average of 100 traces). <i>Right</i>, time course of the LFP amplitude changes (mean±SEM). Drug microperfusion is indicated by a bar and TSS by an arrow.</p

Evidence for presynaptic expression of nicotine facilitated LTP.

<p>Percent changes induced by 10-pulse bursts in EPSC amplitude, release probability (<i>p</i>), coefficient of variation (<i>CV</i>), and failure rate (<i>FR</i>) between 15 and 20 min after the induction of plasticity in the experiments A-C of Fig. 2 (p<0.01 for all parameters).</p