The effects of chloride dynamics on substantia nigra pars reticulata responses to pallidal and striatal inputs - PubMed
- ️Wed Jan 01 2020
The effects of chloride dynamics on substantia nigra pars reticulata responses to pallidal and striatal inputs
Ryan S Phillips et al. Elife. 2020.
Abstract
As a rodent basal ganglia (BG) output nucleus, the substantia nigra pars reticulata (SNr) is well positioned to impact behavior. SNr neurons receive GABAergic inputs from the striatum (direct pathway) and globus pallidus (GPe, indirect pathway). Dominant theories of action selection rely on these pathways' inhibitory actions. Yet, experimental results on SNr responses to these inputs are limited and include excitatory effects. Our study combines experimental and computational work to characterize, explain, and make predictions about these pathways. We observe diverse SNr responses to stimulation of SNr-projecting striatal and GPe neurons, including biphasic and excitatory effects, which our modeling shows can be explained by intracellular chloride processing. Our work predicts that ongoing GPe activity could tune the SNr operating mode, including its responses in decision-making scenarios, and GPe output may modulate synchrony and low-frequency oscillations of SNr neurons, which we confirm using optogenetic stimulation of GPe terminals within the SNr.
Keywords: GABA; chloride homeostasis; decision-making; mouse; neuroscience; oscillations; substantia nigra; synchrony.
© 2020, Phillips et al.
Conflict of interest statement
RP, IR, JR No competing interests declared, AG Reviewing editor, eLife
Figures

(A) Schematic diagram of the model. (B) Tonic spiking voltage traces for both compartments, with minimum voltages labeled. (C) Model f-I curve. (D) Phase plot of the rate of change of the membrane potential (dVm/dt) against the membrane potential (Vm) showing afterhyperpolarization (AHP) and spike height (AP Peak) for both compartments.

For the GPe and Str simulations, the left traces (A and C) show current and right panels (B and D) show the pared pulse ratios (PPR) resulting from repeated synaptic stimulation at different frequencies. The amplitude of each IPSC (Pn) was normalized to the amplitude of the first evoked IPSC (P1). For this set of simulations the membrane potential was held at VS=-60.0mV and EGABA for the somatic and dendritic compartments was held fixed at –72 mV. Model parameters and behavior were tuned to match voltage-clamp data from Connelly et al., 2010.

For the GPe and Str simulations, the left traces (A and C) show voltage and right panels (B and D) show the pared pulse ratios (PPR) resulting from repeated synaptic stimulation at different frequencies. The amplitude of each IPSP (Pn) was normalized to the amplitude of the first evoked IPSP (P1). For this set of simulations IAPP was tuned to set the resting membrane of the somatic compartment at VS=-60.0mV. In both compartments EGABA was held fixed at –70 mV. Model performance is qualitatively, and somewhat quantitatively, similar to experimental current-clamp data (Lavian and Korngreen, 2016, Figures 2, 3).

(A) Dependence of somatic EGABA on the tonic chloride conductance (gGABATonic) and the potassium-chloride co-transporter KCC2 extrusion capacity (gKCC2). (B–E) Examples of SNr responses to simulated indirect pathway stimulation at different positions in the 2D (gGABATonic,gKCC2) parameter space, as labeled in panel (A). (E1 and E2) Notice the two distinct types of partial inhibition. Inset highlights the drift in EGABA during stimulation.

(A) Dependence of somatic EGABA on the tonic chloride conductance (gGABATonic) and the potassium-chloride co-transporter KCC2 extrusion capacity (gKCC2) as previously shown in Figure 4A. (B) Example of ‘Partial Inhibition’ in response to 1 s of stimulation resulting due to a small accumulation of intracellular Cl-, as shown in Figure 4E2. (C) Example of longer 10 s stimulation resulting in larger Cl- accumulation resulting in a transition of EGABA from inhibitory to excitatory and thus causing a biphasic response in a simulated SNr neuron’s firing rate, as seen in 10 s stimulation experiments. gGABATonic and gKCC2 are the same in (B) and (C) and take the values indicated in (A). To generate the biphasic example the synaptic conductance gGABAS was increased from 0.2 to 0.4 nS/pF.

(A) Dependence of somatic EGABA on the tonic chloride conductance (gGABATonic) and the potassium-chloride co-transporter KCC2 extrusion capacity (gKCC2). (B–F) Examples of SNr responses to simulated indirect pathway stimulation at different locations in the 2D (gGABATonic,gKCC2) parameter space. gGABATonic and gKCC2 for each example are indicated in panel (A). (E1 and E2) Notice the two distinct types of partial inhibition. Inset highlights the drift in EGABA during stimulation. (F) Example of a biphasic inhibition-to-excitation response elicited by increasing the stimulation frequency to 40 Hz under the same conditions shown in E2. Alternatively, same response could be elicited by increasing the synaptic weight (WGABAStr). .

(A1 and B1) Examples of response types observed for 10 s stimulation of GPe or Str projections. (A2 and B2) Quantification types of SNr response to optogenetic stimulation at varying frequencies (GPe: n=4 animals, 12 slices, 10 Hz, 20 Hz, and 40 Hz = 40 cells, 60 Hz = 39 cells; Str: n=4 animals, 12 slices, 10 Hz, 20 Hz, and 40 Hz = 33 cells, 60 Hz = 31 cells). (A3 and B3) Effect of GPe or Str stimulation on the firing rate of SNr neurons averaged across all trials for stimulation at 40 Hz. Error bars indicate the standard deviation. The 10 s stimulation period was broken into 1 s intervals to show the gradual weakening of inhibition during stimulation.

(A1–A4) Raster plots of spiking sorted by the duration of the pause in spiking at the start of the stimulation period for all SNr neurons tested. (B1–B4) Effect of GPe stimulation on the firing rate of SNr neurons averaged across all neurons and each stimulation frequencies tested. Error bars indicate SD. (C1 and C2) Quantification of types of SNr responses to optogenetic stimulation for varying frequency characterized in the first (C1) 1 s or the full (C2) 10 s. Notice that fewer neurons are completely inhibited in the full 10s period and some biphasic responses emerge.Str: n=4 animals, 12 slices, 10 Hz, 20 Hz, and 40 Hz = 33 cells, 60 Hz = 31 cells.

(A1–A4) Raster plots of spiking sorted by the duration of the pause in spiking at the start of the stimulation period for all SNr neurons tested. (B1–B4) Effect of Str stimulation on the firing rate of SNr neurons averaged across all neurons and each stimulation frequencies tested. Error bars indicate SD. (C1 and C2) Quantification of types of SNr responses to optogenetic stimulation for varying frequency characterized in the first (C1) 1 s or the full (C2) 10 s. Notice the decrease in the number of completely inhibited neurons and increase in the number of biphasic responses in the full 10 s period. Str: n=4 animals, 12 slices, 10 Hz, 20 Hz, and 40 Hz = 33 cells, 60 Hz = 31 cells.

(A and B) Example traces illustrating the effect of a single GABAergic synaptic input on the phase of spiking in a simulated SNr neuron for hyperpolarized and depolarized EGABA, respectively. (C) For an ongoing voltage oscillation of a spiking SNr neuron (blue trace), we define a phase variable as progressing from 0 immediately after a spike to one at the peak of a spike. As EGABA is varied from –60 mV to –50 mV, progressively more of the SNr voltage trace lies below EGABA, where GABAergic inputs have depolarizing effects. (D) PRCs computed for a model SNr neuron in response to GABAergic input stimuli arriving at different phases of an ongoing SNr oscillation. As EGABA is varied from –60 mV to –50 mV, the PRC transitions from a curve showing a delay of the next spike for most stimulus arrival phases, through some biphasic regimes, to a curve showing an advance of the next spike for almost all possible phases. In panel D, the A and B labels at approximately 0.5 phase on the EGABA=-60mV and EGABA=-50mV PRCs correspond to the examples shown in panels A and B. The conductance of the synaptic input was fixed at 0.1 nS/pF in order to produce deflections in Vm for hyperpolarized EGABA that are consistent with data presented in Higgs and Wilson, 2016.

(A1-A3 and B1-B3) (Top) Identification of PRC fixed points and (Bottom) histogram of the timing of synaptic inputs in the phase of neuron 2 (Input Phase) as a function of EGABA. Recall that positive changes in phase correspond to delays. (A1,B1) EGABA=-60mV; (A2,B2) EGABA=-55mV; (A3,B3) EGABA=-50mV. Black dots indicate dataset used to generate PRC in red/blue. Stable and unstable fixed points are indicated by green and white filled circles, respectively. For reference, all PRCs and fixed points are included in gray for all values of EGABA tested. (A4,B4) Effect of EGABA on SNr phase locking. Blue histograms show the distribution of synaptic inputs relative to the phase of neuron 2 (input phase) for the two network simulations for different levels of EGABA. Green and white filled circles indicate the stable and unstable locking predicted by analysis of PRCs. Note the unstable fixed points for the lowest values of EGABA in the unidirectional case. (A5) In the unidirectional case, slow 1 Hz oscillations in the frequency of neuron 2 arise due to phase slipping at hyperpolarized values of EGABA.

Each vertical, deeply colored bar denotes a spike time of the cell with that color (red or blue). Following the spike time of each cell, the PRC for that cell is shown (the PRCs for EGABA=-60mV are used). The spike time of one cell becomes the input time to the other, target cell (‘Input’). The height of the PRC for the target cell at the arrival time of its input determines the delay (‘Delay’) until its next spike. Each pale vertical bar shows the time when that next spike would have occurred had the corresponding cell not received an input; each horizontal arrow shows how the delay due to input perturbs the cell’s actual next spike time and determines the timing of the next input to the other cell. Over successive spikes and delays, the relative phases of the cells drift, such that the cells’ spike times approach the fully anti-phase locked state, in which each cell spikes, and sends input to the other cell, half-way through each interspike interval.

(A) Illustration of the phase of the postsynaptic neuron at the moment when it receives each input from the presynaptic neuron (input phase) for the unidirectionally connected two neuron network as a function of time for EGABA=-60mV. Dots denote phases of the postsynaptic neuron when the presynaptic neuron spikes. The phase value of 1 corresponds to the postsynaptic neuron being at spike threshold. Insets show the timing of the presynaptic neuron spike (black triangle/dashed line), the phase of the postsynaptic neuron spike after it receives the input (blue), and the spike train of the postsynaptic neuron in the absence of input (gray). The red cycle is used in B. (B) Overlay of the PRC generated for EGABA=-60mV and the resulting progression of phase for one full phase slipping oscillation. Light gray dots indicate the data points used to generate the blue PRC. Recall that positive changes in phase correspond to delays. (C) The frequency of phase slipping increases as EGABA decreases, with a steeper relationship for larger synaptic weight (WGABASNr) between the SNr neurons. .

(A) Tuning curve for presynaptic firing rate (FR) versus applied current, IAPP. Dashed line indicates the baseline firing rate (10.5 Hz) with no applied current. (B) Histograms of the input phase in the postsynaptic neuron under baseline conditions (IAPP=0pA/pF). (C1–C4) Input phase histograms for different presynaptic firing rates (9.76 Hz, 10.15 Hz, 10.91 Hz, 11.26 Hz from left to right). EGABA ranges are not the same in all panels. Regions of phase slipping (PS) and phase advancing (PA) oscillations are each indicated by a solid horizontal bar. Example oscillations in input phase (D1–D4) and postsynaptic firing rate (E1–E4) at specific values of EGABA for different presynaptic firing rates highlighted in red for the corresponding D panels. Notice that the PA oscillations in C1-2 and D1-2 result in periodic increases in the postsynaptic firing rate in E1-2 whereas PS oscillations in C3-4 and D3-4 result in periodic decreases in firing rate in E3-4.

(A) Relationship between applied current (IAPP) and the firing rate of an isolated SNr model neuron. (B) Example voltage trace for a simulated neuron firing at 33.0 Hz with IAPP=0.8pA/pF. (C) Histograms of input phase in the two SNr neurons with unidirectional (feed forward) connectivity as a function of EGABA. Oscillations occur for EGABA≤-57mV and strict phase locking for EGABA>-57mV. (D) Example plot showing slow oscillations in the phase of the presynaptic neuron at which the postsynaptic spike occurs (input phase) over time. (E) Example slow oscillations in the instantaneous firing rate of the postsynaptic neuron. EGABA=-60mV in panels D and E.

(A) Dependence of CV (blue) and firing rate (red) of model SNr neuron on applied noise amplitude. (B) Example voltage trace at the highest level of added Gaussian noise (0.6pA/pF) and CV=0.1. (C) Relationships between EGABA and input phase distributions for increasing Gaussian noise: (C1) 0.2pA/pF, (C2) 0.4pA/pF, and (c3) 0.6pA/pF. (D–E) Example slow oscillations in the input phase (D1–D3) and the post synaptic firing rate (E1–E3). EGABA values in the example traces are indicated by red histograms in the corresponding panel in (C). (F1) Example power spectrum curves in the 0–3 Hz range with 0.0, 0.2, 0.4, and 0.6 pA/pf of Gaussian noise and EGABA=-60mV. In each case, results shown are remaining power after subtraction of the mean power in the 3–4 Hz range. (G) Area of the power spectrum peak as a function of EGABA and applied Gaussian noise. (H) CV in the postsynaptic neuron as a function of EGABA and added Gaussian noise. Note that postsynaptic CVs exceed those reported in the literature when we take a noise amplitude of 0.6 pA/pf, suggesting that this value may be excessive.

(A–C) Examples of synaptic delays of increasing magnitude: 0 mS, 1.6 mS, and 8.6 mS, respectively. (D) Histogram of input phase in the postsynaptic neuron as a function of EGABA and varying delay duration. Notice that delays do not strongly effect the neurons’ spiking relationship.

(A) Characterization of the varying degrees of presynaptic synchrony defined by the parameter presynaptic offset. The presynaptic offset is the phase difference between the first (red) and second (blue) presynaptic neuron. (B and C) Effect of varying presynaptic offset on postsynaptic (B) firing rate and (C) coefficient of variation (CV). (D1–D6) Input phase histograms for synaptic inputs in the postsynaptic neuron from the first presynaptic neuron as a function of the presynaptic offset and EGABA. (E1–E6) Power spectrum in the post synaptic neurons as a function of the presynaptic offset and EGABA.

(A) Characterization of the varying degrees of presynaptic synchrony defined by the parameter presynaptic offset. The presynaptic offset is the phase difference between the first (red),second (green) and third (blue) presynaptic neuron. (B and C) Effect of varying presynaptic offset on postsynaptic (B) firing rate and (C) coefficient of variation (CV). (D1–D6) Input phase histograms for synaptic inputs in the postsynaptic neuron from the first presynaptic neuron as a function of the presynaptic offset and EGABA. (E1–E6) Power spectrum in the post synaptic neurons as a function of the presynaptic offset and EGABA.

(A) Characterization of the varying degrees of presynaptic synchrony defined by the parameter presynaptic offset. The presynaptic offset is the phase difference between the first (red),second (green),third (blue) and fourth (purple) presynaptic neuron. (B and C) Effect of varying presynaptic offset on postsynaptic (B) firing rate and (C) coefficient of variation (CV). (D1–D6) Input phase histograms for synaptic inputs in the postsynaptic neuron from the first presynaptic neuron as a function of the presynaptic offset and EGABA. (E1–E6) Power spectrum in the post synaptic neurons as a function of the presynaptic offset and EGABA.

(A) Phase difference between the two presynaptic neurons (cell 1 and cell 2) as a function of EGABA. (B) Firing rate and coefficient of variation (CV) in the postsynaptic neuron (cell 3) as a function of EGABA. (C) Input phase histograms for synaptic inputs in the postsynaptic neuron from the first presynaptic neuron as a function of EGABA. (D) Power spectrum in the postsynaptic neuron as a function of EGABA. (E1 and E2) Example traces of the input phase relationship between the postsynaptic neuron and the first presynaptic neuron. The value of EGABA is indicated to the left of each trace.

(A) Histogram showing the number of neurons receiving zero to eight synaptic inputs. (B) Mean network firing rate as a function of EGABA. (C) Mean network CV as a function of EGABA. Shaded regions in B and C represent standard deviation. (D1–D3) Example raster plots of spikes in the network for three different values of EGABA. (E1–E3) Power spectrum for each neuron in the network for the same three values of EGABA used in (D1–D3). Rows in D and E panels are sorted by the number of inputs from least (cell 1) to most (cell 100). Green arrows point out peaks in the power spectra of example neurons examined in the following panels. (F1 and F2) Left panel: Power spectrum for two example neurons with peaks indicating slow oscillations. Right panels: slow oscillation in the instantaneous firing rate in the two example neurons. These are shown for EGABA=-60mV (F1) and for EGABA=-50mV (F2).

(A–B) Example (A) raster plot and (B) power spectrum of a single spiking unit in SNr without (blue) and with (red) optogenetic stimulation of GPe terminals over multiple trials. (C) Frequencies of slow oscillations in the 12 unit dataset before optogenetic stimulation. (D) Distribution of single unit firing rates without (blue) and with (red) optogenetic stimulation for all recorded units (n = 12). Notice that stimulation has no significant effect on firing rate (t-test p=0.8531). (E) Band power (0.75–3.0 Hz) without (blue) and with (red) optogenetic stimulation for oscillatory units (n = 5). Solid blue and red horizontal bars indicate mean band power. Notice that stimulation significantly reduces the power of the slow oscillations (t-test p=0.0341). Recordings were collected from four animals. .

(A) Raster plot of spikes in the simulation of an SNr network model containing 50 simulated neurons that receive tonic somatic inhibition from GPe projections. (B) Integrated SNr population activity gives a mean firing rate of about 23 Hz, as seen in in vivo conditions (Freeze et al., 2013; Mastro et al., 2017; Willard et al., 2019). (C) Increasing tonic Cl- depolarizes somatic and dendritic EGABA. (D) Ramping Str synaptic inputs used to represent evidence accumulation in a perceptual decision-making task. (E) Inhibition and pause generation in the SNr during evidence accumulation/ramping Str activity, for two different tonic somatic Cl- conductances. (F) Increasing the tonic Cl- conductance lengthens Tpause, the time for the SNr firing rate to drop below threshold (colors correspond to threshold levels in E). If the tonic conductance becomes too great, then SNr firing cannot be pushed to arbitrarily low rates. .

Neuronal response (blue) and EGABA dynamics (red) in a neuron with (A,C) two or (B,D) four thin dendrites. For comparison EGABA dynamics in a neuron with a single dendrite is shown in gray in all panels. The total capacitance and surface area of the dendritic compartments in A and B matched that of the single dendrite; the total volume of the dendritic compartments in C and D matched that of the single dendrite. The stimulation period is from 0–1 s and is indicated by the horizontal black bar.

Relationship between the tonic Cl- conductance and Tpause for (A) one dendrite, (B) two (B1) or four (B2) dendrites with total surface area and capacitance matched to the single dendrite, and (C) two (C1) or four (C2) dendrites with total volume matched to the single dendrite. Colors correspond to three different pause thresholds, defined by SNr firing rates, as in Figure 11E in the main manuscript.

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