Pupil diameter is not an accurate real-time readout of locus coeruleus activity
- Department of Molecular, Cell and Systems Biology, University of California, Riverside, United States
- Neuroscience Graduate Program, University of California, Riverside, United States
Figures
Figure 1 with 5 supplements
Correlating locus coeruleus (LC) activity to pupil responses.
(a) Left: schematic of experimental setup for simultaneous pupil and LC recording/optical stimulation in head-fixed mice. Lightning bolt: light pulse. Right: expression of ChR2 in a DBH;Ai32 mouse …
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Figure 1—source data 1
Source data for Figure 1.
- https://cdn.elifesciences.org/articles/70510/elife-70510-fig1-data1-v1.zip
Figure 1—figure supplement 1
Locus coeruleus (LC) and pupil recordings in mice.
(a) Distribution of interspike interval, spike waveform (top) and spike sorting diagram (bottom) of three example recordings with median interspike interval (ISI, left to right: 0.20, 0.29, and 0.36 …
Figure 1—figure supplement 2
The relationship between the latency of peak pupil diameter and locus coeruleus (LC) spike cluster size.
c.c., Pearson correlation coefficient.
Figure 1—figure supplement 3
The relationship between pupil size changes and locus coeruleus (LC) spike cluster.
The relationship between pupil size changes and LC spike cluster when pupil responses were quantified as % changes from baseline (a), time derivative (b), or using a shorter time window (3 s, c). …
Figure 1—figure supplement 4
Group mean area under the curve (AUC) values when using peak pupil diameter to predict the associated cluster sizes 1 through 8 from all recordings (n = 19).
Figure 1—figure supplement 5
Group mean probability distribution of locus coeruleus (LC) spike clusters (n = 19).
Figure 2 with 2 supplements
Reverse correlating pupil responses to locus coeruleus (LC) activity.
(a) Example pupil–LC traces showing the detected pupil dilation events (blue arrows) based on zero-crossing of pupil derivatives. (b) Probability distributions of LC spike counts associated with …
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Figure 2—source data 1
Source data for Figure 2.
- https://cdn.elifesciences.org/articles/70510/elife-70510-fig2-data1-v1.zip
Figure 2—figure supplement 1
Group mean relationship between peak pupil diameter and locus coeruleus (LC) spike counts using two different methods.
Group mean relationship between peak pupil diameter and LC spike counts based on (1) clustering LC spikes then identifying the associated peak pupil responses (blue, Figure 1), and (2) detecting …
Figure 2—figure supplement 2
Group mean probability distribution of the detected pupil dilation events (n = 19).
Pupil dilation events were binned every 0.3 standard deviation (SD).
Figure 3 with 1 supplement
Locus coeruleus (LC) responses to optogenetic stimulation.
(a) Example LC histological section illustrating optical fiber implant and the estimated excitable volume (light gray cone). Estimates were based on 10-mW laser power, 2.5 mW/mm2 excitation …
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Figure 3—source data 1
Source data for Figure 3.
- https://cdn.elifesciences.org/articles/70510/elife-70510-fig3-data1-v1.zip
Figure 3—figure supplement 1
Locus coeruleus (LC) response to optogenetic stimulation.
The number of evoked spikes from an opto-tagged LC unit in response to optical stimulation using different laser power (single 10 ms pulse; left), or different pulse width (single pulse; right). …
Figure 4 with 6 supplements
Pupil responses to locus coeruleus (LC) optogenetic stimulation.
(a) Example responses from the same pupil to LC stimulation in two awake, baseline pupil-matched sessions (left and right) aligned to the onset of optical stimulation of four 10 ms pulses at 10 Hz. …
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Figure 4—source data 1
Source data for Figure 4.
- https://cdn.elifesciences.org/articles/70510/elife-70510-fig4-data1-v1.zip
Figure 4—figure supplement 1
Raw pupil traces for the two sessions used in Figure 4a, b.
Figure 4—figure supplement 2
Unnormalized group pupil responses as shown in Figure 4c, d.
Session-to-session fluctuations were not observable from group comparisons. p = 0.38, n = 12 for (a) and p = 0.63, n = 11 for (b). Magenta arrows indicate same-day comparison.
Figure 4—figure supplement 3
The variability of pupil responses to locus coeruleus (LC) optical stimulation within individual sessions (Within) was comparable to that of across sessions (Across) in awake mice.
(a) p = 0.53, p = 0.58. (b) p = 0.22, p = 0.15. Two-tailed rank sum test. Across-session variability was estimated by resampling pooled trials from all sessions in each condition. The iteration of …
Figure 4—figure supplement 4
Spontaneous pupil fluctuation was reduced during anesthesia.
(a) The amplitude of spontaneous pupil dilation events in awake, nontask performing condition was larger than in anesthetized condition (five sessions from three mice in each condition, p = 0.0079, …
Figure 4—figure supplement 5
Simultaneous locus coeruleus (LC) and pupil responses to optical stimulation.
Simultaneous recording of an LC unit waveform (a), spike responses (b), and pupil diameter (c) during optogenetic stimulation (10 ms pulse train: four pulses at 10 Hz [top] and eight pulses at 5 Hz …
Figure 4—figure supplement 6
List of all locus coeruleus (LC) recordings.
(a, b) ISI distribution and spike waveform from all recordings, ranked by increasing median ISI. Pupil–LC relationship with R2 > 0.6 (as in Figure 1e) in (a, n = 13), and the remaining in (b, n = …
Figure 5
Pupil–locus coeruleus (LC) coupling correlated with decision-bias-related variables.
(a) The variations in the relationship between peak pupil diameter and LC cluster size (linear slopes in Figure 1e) were strongly correlated with Hit rate (left) and decision bias (right, n = 9). …
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Figure 5—source data 1
Source data for Figure 5.
- https://cdn.elifesciences.org/articles/70510/elife-70510-fig5-data1-v1.zip
Author response image 1
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (M. musculus) | DBH-Cre | MMRRC | RRID:MMRRC_036778-UCD | |
| Strain, strain background (M. musculus) | Ai32 | JAX | RRID:IMSR_JAX:012569 | |
| Software, algorithm | BControl | Princeton University | https://brodylabwiki.princeton.edu/bcontrol | |
| Software, algorithm | WaveSurfer | HHMI Janelia | http://wavesurfer.janelia.org/ | |
| Software, algorithm | Matlab | MathWorks | RRID:SCR_001622 | |
| Software, algorithm | Janelia eye tracker | HHMI Janelia | N/A | |
| Software, algorithm | StreamPix | Norpix | RRID:SCR_015773 | |
| Software, algorithm | Illustrator | Adobe | RRID:SCR_010279 | |
| Other | Camera | PhotonFocus | DR1-D1312-200-G2-8 | |
| Other | Telecentric lens | Edmund Optics | 55–349 | |
| Other | Tetrode drive | Cohen et al., 2012 | N/A | |
| Antibody | Anti-TH primary antibody | Thermo Fisher | OPA104050 RRID:AB_325653 | 1:1000 |
| Antibody | Secondary antibody | Thermo Fisher | A11008 RRID:AB_2534079 | 1:500 |
