How inhibitory and excitatory inputs gate output of the inferior olive
- Department of Neuroscience, Erasmus MC, Netherlands
- Netherlands Institute for Neuroscience, Royal Academy of Arts & Sciences, Netherlands
- Faculty of Health Sciences, Universidad Católica Silva Henríquez, Chile
Figures
Figure 1
In vitro whole-cell patch-clamp recordings of oscillating and non-oscillating neurons in the inferior olive (IO).
(A) Dye coupling between electrically coupled neurons in the IO was visualized by performing immunostaining on neurons dialyzed with biocytin. Note that the dendrites of the cluster of the labeled …
Figure 2 with 2 supplements
Responses of oscillating and non-oscillating neurons in the inferior olive (IO) to stimulation of GABAergic afferents from the cerebellar nuclei (CN).
(A) Either we expressed ChrimsonR-tdTomato in the CN of wild type mice or we use GAD2Cre/Chr2- H134R-EYFP transgenic mice so as to be able to specifically stimulate the GABAergic input to the IO …
Figure 2—figure supplement 1
Picrotoxin blocks the cerebellar nuclei (CN) synaptic response.
(A) Example trace of synaptic response to CN inputs before and after the presence of picrotoxin (PTX, 100 µM). (B) Vhyp and Vreb normalized amplitude before (baseline) and after the presence of PTX. …
Figure 2—figure supplement 2
Voltage dependence of the rebound depolarization (Vreb) amplitude of cerebellar nuclei (CN) and mesodiencephalic junction (MDJ) synaptic responses.
(A) Vreb amplitude (mV) of CN synaptic response plotted as a function of membrane voltage (Vm) previous to stimulation. Open purple circles represent individual trials. (B) Vreb amplitude (mV) of CN …
Figure 3 with 1 supplement
Responses of oscillating and non-oscillating neurons in the inferior olive to stimulation of excitatory afferents from the mesodiencephalic junction (MDJ).
(A) To stimulate neurons in the principal olive (PO) and medial accessory olive (MAO), we used either direct electrical stimulation of the central and medial tegmental tract, respectively (top …
Figure 3—figure supplement 1
Mesodiencephalic junction (MDJ) evoked synaptic responses in the inferior olive (IO).
(A) Synaptic responses to increasing stimulus intensities using electrical stimulation. Arrows indicate how depolarizing (Vdep), hyperpolarizing (Vhyp), and rebound (Vreb) synaptic components …
Figure 4
Comparison of subthreshold responses of oscillating olivary neurons obtained with experiments (data) and those found following simulations (model).
(A) Subthreshold oscillatory responses to stimulation of the cerebellar nuclei (onset indicated by pink arrow-heads). Panels on the left and right show subthreshold activity according to the data …
Figure 5 with 1 supplement
Timing of input from mesodiencephalic junction (MDJ) with respect to that from the cerebellar nuclei (CN) is critical for olivary output.
(A) To stimulate the excitatory input from the MDJ to neurons in the principal olive (PO) and medial accessory olive (MAO), we used either optogenetic stimulation using Chronos-eYFP (top) or direct …
Figure 5—figure supplement 1
Phase response curve (PRC) calculation.
(A) Period previous to stimulation (T0) was calculated as the average of four periods preceding the stimuli. Period containing the synaptic responses caused by either single or dual stimuli (T1) was …
Figure 6
Impact of cerebellar nuclei (CN), mesodiencephalic junction (MDJ), and combined CN-MDJ stimulation on suprathreshold responses of inferior olive (IO) oscillating neurons.
(A) Suprathreshold responses in IO neurons evoked by CN stimulation. In the left panel CN stimulus onset is indicated by a pink arrow-head, while action potentials are clipped in order to visualize …
Figure 7
Modeling indicates that resetting of subthreshold activity and spike generation are more effective at the preferred interval.
A single model example cell (with a Ca v.3.1 conductance of 1.1 mS/cm2) is stimulated at different phases of the oscillation. Stimulations were delivered at 16 different moments within one cycle. (A)…
Figure 8 with 6 supplements
Network model of inferior olive can recapitulate timing-sensitive interaction between inhibitory and excitatory inputs.
Combined stimulation at a preferred interval of about one cycle leads to highest rebound and maintenance of network synchronization. (A) Data from model of the olivary neuropil based on a …
Figure 8—figure supplement 1
Cell conductances parameter spaces for coupled and uncoupled networks show effect of gap junction coupling in murine scale model network.
Networks generated for this project have high levels of randomization to capture expected experimental variability. All 1105 cells in the model receive randomized conductance parameters. Displayed …
Figure 8—figure supplement 2
Stimulation at non-preferred intervals leads to enduring phase differences.
(A) Example phase average of stimulated group and non-stimulated network. The mesodiencephalic junction (MDJ) signal causes momentary desynchronization of the stimulated group. After the cerebellar …
Figure 8—figure supplement 3
Responses of network to combined stimulation under noise.
Due to its incoming connections form a wide array of excitatory sources, the inferior olive is subjected to highly variable inputs (Negrello et al., 2019). When olivary neurons are given noisy …
Figure 8—video 1
Combined stimulation can lead to standing wave propagations.
Displayed is phase of the oscillation for all cells in network (blue is 0 pi and red is 2 pi). The combined impact of inhibition and excitation and the ongoing subthreshold oscillations creates …
Figure 8—video 2
Different stimulation intervals lead to systematic phase differences.
Top row: phase distributions displaying whole network (in gray) and stimulated population (violet). Bottom row, phase distributions over time for stimulated group (top) and rest of the network …
Figure 8—video 3
Oscillator phase in the reconstructed olive during the presence of Ornstein-Ulhenbeck noise.
Even though conjoined stimulation (AMPA input follows GABA input after 110 ms) drives phase reset, this effect washes off in about two cycles. Noise parameters: theta = 1/50 ms, mean = 0 pA/cm2, std …
Figure 9 with 1 supplement
Network responses to combined stimulation for different intervals show an increase in rebound amplitude as well as an increase in the proportion of oscillating cells following stimulation in the preferred interval.
(A) Average of membrane potential for entire network for the preferred network stimulus (100 ms). (B) Collected response snippets for a variety of stimulation intervals. Dashed inset indicates the …
Figure 9—figure supplement 1
Network tuning for calcium conductance to capture proportion of oscillating cells.
The main determinant of oscillation amplitude is calcium low-threshold type (v 3.1). To obtain a network with the correct proportion of oscillating cells, we have produced five networks with …
Author response image 1
Author response image 2
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | C57BL/6J mice | Charles Rivers | IMSR_JAX:000664 | |
| Strain, strain background (Mus musculus) | Gad2-IRES-Cre × Ai32 RCL-ChR2(H134R)/EYFP | https://doi.org/10.1073/pnas.1017210108; https://doi.org/10.1038/nature13276 | ||
| Transfected construct (AAV1-Synapsin-Chronos-GFP) | Chronos | https://doi.org/10.1038/nmeth.2836 | ||
| Transfected construct (AAV1-Synapsin-ChrimsonR-tdTomato) | Chrimson | https://doi.org/10.1038/nmeth.2836 | ||
| Chemical compound, drug | Alexa Fluor 594- Conjugated Streptavidin | Life Technologies | Stock concentration 2 mg/ml (1:200 dilution) | |
| Chemical compound, drug | Alexa Fluor 633- Conjugated Streptavidin | Life Technologies | Stock concentration 2 mg/ml (1:200 dilution) | |
| Chemical compound, drug | APV | Hello Bio | #HB0225-50mM | A final concentration of 50 µM was used |
| Chemical compound, drug | CNQX | Hello Bio | #HB0205-50mg | A final concentration of 20 µM was used |
| Chemical compound, drug | Picrotoxin (PTX) | Sigma-Aldrich | #P1675-5G | A final concentration of 100 µM was used |
| Chemical compound, drug | Biocytin | Sigma-Aldrich | #B4261-50MG | A final concentration of 0.5% (wt/vol) was used |
| Software, algorithm | Clampfit 11.0.3 | Molecular Devices | ||
| Software, algorithm | Prism 9 | GraphPad | ||
| Software, algorithm | MATLAB | MathWorks |
Additional files
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Supplementary file 1
Reponse properties of neurons in the inferrior olive.
(A) Kinetic properties of cerebellar nuclei (CN) and mesodiencephalic junction (MDJ) synaptic responses using different stimulation approaches. Latency and half-width of hyperpolarization (Vhyp) and rebound (Vreb) synaptic components of CN synaptic responses evoked in mice transduced with ChrimsonR-tdTomato in the CN and GAD2Cre/Chr2-H134R-EYFP transgenic mice line, and latency and half-width of depolarization (Vdep), hyperpolarization (Vhyp), and rebound (Vreb) synaptic components of MDJ synaptic response evoked in mice transduced with Chronos in the MDJ and electrical stimulation. (B) Phase response curve (PRC) parameters of different stimulation paradigms. Slope, Y-intercept, and R2 of PRCs generated by dual (CN and MDJ afferents stimulation) stimulation at different time intervals (all of them evoking subthreshold synaptic responses except for ‘+150 ms spike’ which evokes suprathreshold synaptic responses), CN afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘IPSP’ and ‘CN spike’, respectively), and MDJ afferent stimulation evoking suprathreshold synaptic responses (‘MDJ spike’). (C) Statistics of PRC slopes of different stimulation paradigms. Comparison of PRC slopes between +150 ms and the other time intervals (all of them evoking subthreshold synaptic responses except for ‘+150 ms spike’ which evokes suprathreshold synaptic responses), CN afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘IPSP’ and ‘CN spike’, respectively) and MDJ afferent stimulation evoking suprathreshold synaptic responses (‘MDJ spike’), using one-way ANOVA test followed by post hoc uncorrected Fisher’s LSD multiple comparison test. (D) Statistics of PRC Y-intercept. Comparison of PRC Y-intercepts between +150 ms and the other time intervals (all of them evoking subthreshold synaptic responses except for ‘+150 ms spike’ which evokes suprathreshold synaptic responses), CN afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘IPSP’ and ‘CN spike’, respectively) and MDJ afferent stimulation evoking suprathreshold synaptic responses (‘MDJ spike’) using one-way ANOVA test followed by post hoc uncorrected Fisher’s LSD multiple comparison test. (E) Intertrial phase jitter at different stimulation paradigms. Intertrial phase jitter pre- and post-stimulus (standard deviation of phase lags pre- and post-stimulus, respectively) and intertrial phase jitter change (standard deviation of phase lags post stimulus – standard deviation of phase lags pre-stimulus) at different time intervals of stimulation (all of them evoking subthreshold synaptic responses except for ‘+150 ms spike’ which evokes suprathreshold synaptic responses), CN afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘IPSP’ and ‘CN spike’, respectively) and MDJ afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘EPSP’ and ‘MDJ spike’, respectively). (F). Statistics of amplitude of first cycle following stimulation at different time intervals. Comparison between subthreshold oscillation (STO) amplitude change of the first cycle (rebound component) following stimulation at different time intervals (all of them evoking subthreshold synaptic responses) and STO amplitude previous to stimulation using Friedman test followed by post hoc uncorrected Dunn’s multiple comparison test. (G) STO amplitude change of first cycle following stimulation at different stimulation paradigms. STO amplitude change of the first cycle (rebound component) following dual stimulation at different time intervals (all of them evoking subthreshold synaptic responses except for ‘+150 ms spike’ which evokes suprathreshold synaptic responses), CN afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘IPSP’ and ‘CN spike’, respectively), and MDJ afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘EPSP’ and ‘MDJ spike’, respectively). (H) Statistics of the comparison between first cycle post-stimulus change evoked by +150 ms and other stimulation paradigms. Comparison of first cycle post-stimulus change between the time interval of +150 ms and the other time intervals (all of them evoking subthreshold synaptic responses except for ‘+150 ms spike’ which evokes suprathreshold synaptic responses), CN afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘IPSP’ and ‘CN spike’, respectively), and MDJ afferent stimulation evoking subthreshold and suprathreshold synaptic responses (‘EPSP’ and ‘MDJ spike’, respectively), using Kruskal-Wallis test followed by post hoc uncorrected Dunn’s multiple comparison test. (I) Spike probability at different stimulation paradigms. Spike probability (Pspike) following dual stimulation at different time intervals and isolated CN and MDJ afferents stimulation (‘IPSP’ and ‘MDJ’, respectively).
- https://cdn.elifesciences.org/articles/83239/elife-83239-supp1-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/83239/elife-83239-mdarchecklist1-v1.pdf
