Functional clustering of dendritic activity during decision-making
- Janelia Research Campus, Howard Hughes Medical Institute, United States
Figures
Figure 1 with 3 supplements
Targeted high-speed imaging in behaving mice.
(A) Optical layout for high-speed, high-resolution imaging in three dimensions. An x-axis mirror galvanometer, remote focusing arm, and prism-based GDD compensation unit were added to a high …
Figure 1—figure supplement 1
Histological characterization of transgenic mouse lines.
(A) GCaMP labeling in an example coronal section of Syt17-Cre x Ai93 approximately 1.5 mm anterior to bregma. (B) GCaMP labeling in an example coronal section of OE25-Cre x Ai93 approximately 1.5 mm …
Figure 1—figure supplement 2
Image regestration example.
(A) Tracing of dendrites (top) and spatiotemporal distribution of imaging frames (bottom) for an example L2/3 cell imaging session. (B) Maximum intensity projection (MIP, 80 μm z-axis) of the …
Figure 1—video 1
Registration Example.
Top, target for registration is in yellow, single timepoints are in blue. Red arrows indicate the calculated shifts per imaging field. Arrows are 3x real size to emphasize the pixel sized shifts. …
Figure 2 with 3 supplements
Dendrite and spine calcium activity.
(A) Mice were trained to lick either a right or left target based on pole location. The pole was within reach of the whiskers during the sample epoch. Mice were trained to withhold licks until after …
Figure 2—figure supplement 1
Examples of branch-specific persistent calcium activity.
(A) Example of branch-specific persistence of calcium activity following multi-branch events in the dendrites of a L2/3 neuron (soma not imaged in this session). Top, example apical trunk (black) …
Figure 2—video 1
Layer 2/3 Example Calcium Activity.
Top, post-registration activity (AU) for the soma (white), an example spine (green circle), and a segment of dendrite (magenta arrow). Bottom, ΔF/F of the soma (white), spine (green), and dendrite …
Figure 2—video 2
Layer 5 Example Calcium Activity.
Top, post-registration activity (AU) for the trunk (white), an example spine (green circle), and a segment of dendrite (magenta arrow). Bottom, ΔF/F of the soma (white), spine (green), and dendrite …
Figure 3 with 2 supplements
Coincidence of dendritic calcium transients with events in the soma and apical trunk.
(A) Estimation of isolated spine activity as a function of threshold. (i) Example spine ΔF/F (green), threshold (red - 100% ∆F/F) and estimated uncertainty in ΔF/F due to shot-noise (gray shading). …
Figure 3—figure supplement 1
Coincidence activity analysis with different thresholds.
(A) Proportion of time that dendritic spines (left heatmaps) or dendritic segments (right heatmaps) are co-active with the bAP reference signal (soma or apical trunk) as a function of arbitrary …
Figure 3—figure supplement 2
Excluding independent activity before or after reference activity.
(A) Illustration of the transformation of reference (black) and dendritic (magenta) fluorescence signals to an estimate of independent activity (see Figure 3A; Materials and methods). (B) To exclude …
Figure 4 with 2 supplements
Local selectivity after removing the bAP component.
(A) Subtraction of the bAP component from spine signals and estimation of trial average responses for two example spines. Image: MIP of an example L2/3 cell. Light lines: ΔF/F for all 110 correct …
Figure 4—figure supplement 1
Process for removing the estimated bAP-component of spine and dendritic segment signals.
(A) Example reference signal (soma; black line). The reference signal was deconvolved by FOOPSI (Pnevmatikakis et al., 2016; Vogelstein et al., 2010) with parameters set to bias towards a dense …
Figure 4—figure supplement 2
Example of independent and trial averaged activity in a L5 tuft dendrite segment.
(A) Top, example apical trunk (black) and two dendritic branches (light and dark magenta) calcium signals. Bottom, maximum intensity projections (au) and branch insets at example times (dashed …
Figure 5 with 1 supplement
Dendrite and spine calcium signals exhibit diverse selectivity for trial epoch and trial type.
(A) Location of simultaneously imaged dendrite (red lines) relative to the soma (black dot) and connecting dendrite (purple) that was not imaged for an example imaging session of a L2/3 cell. (B) …
Figure 5—figure supplement 1
Dendrite Signals Exhibit Diverse Selectivity for Trial Epoch and Trial Type.
(A) Distribution for L2/3 neurons of differences in epoch selectivity between epoch selective dendrite segments of 3 um (epoch angle CI <30 degrees) on the same neuron for L2/3 neurons (N = 28389 …
Figure 6 with 2 supplements
Interpretation of dendritic calcium signals before and after removal of the estimated contribution from back-propagating action potentials (bAPs).
(A) Spine responses under different bAP subtraction regimes. Top, Cartoon of the soma and spatial organization of five spines. Soma trial-average response (black curve) is centered between the …
Figure 6—figure supplement 1
Simulating the effects of removing the estimated bAP-component on the inferred correlation between spines and the reference signal.
(A) Conceptual diagram of three processes for simulating fluorescence at spines from pre- and post-synaptic spiking. Traces are normalized to maximum. (B) Illustration of how the temporal structure …
Figure 6—figure supplement 2
Simulating the effects of removing the estimated bAP-component on the inferred length constant of correlation between spines.
(A) Illustration of simulating pre-synaptic clustering. Correlation in the spiking of pre-synaptic input varied as a function of distance between the inputs along the dendrite. (B) Illustration of …
Figure 7
Behavior-related calcium signals are organized in a distance-dependent manner within the dendritic tree.
(A) Estimation of pairwise signal correlation after bAP subtraction for two example spines (denoted in (B)). Trial average responses for each epoch and trial type are calculated for each mask from …
Figure 8
Dendritic branching compartmentalizes behavior-related calcium signals.
(A) Example of clustering of epoch selectivity for a L2/3 session. Hue and saturation were determined for each mask as in Figure 5B after bAP subtraction. Markers (gray: individual masks, black: …
Author response image 1
Correlation between decay, amplitude, and proportion independent.
Each point is a 30 μm segment of a L5 tuft dendrite.
Videos
Video 1
Exploring the data online with SpineVis.
In this screencast we show how to use the SpineVis website to look at the data in Figure 2B. On top center is the main viewing area where dragging will change the view in 3D. Clicking on a mask in …
Tables
Table 1
SpineVis online viewer links.
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (M. musculus) | Syt17 NO14 (Mouse) | GENSAT | MMRRC Cat# 034355-UCD, RRID: MMRRC_034355-UCD | |
| Genetic reagent (M. musculus) | CamK2a-tTA | JAX | IMSR Cat# JAX:007004, RRID: IMSR_JAX:007004 | |
| Genetic reagent (M. musculus) | Ai93 | JAX | IMSR Cat# JAX:024103, RRID: IMSR_JAX:024103 | |
| Genetic reagent (M. musculus) | Chrna2 OE25 | GENSAT | MMRRC Cat# 036502-UCD, RRID: MMRRC_036502-UCD | |
| Genetic reagent (M. musculus) | ZtTA | JAX | IMSR Cat# JAX: 012266, RRID: IMSR_JAX:012266 | |
| Software and Algorithms | Matlab | Mathworks | RRID:SCR_001622 | |
| Software and Algorithms | ScanImage | Vidrio | RRID:SCR_014307 | |
| Software and Algorithms | Neuromantic | University of Reading | RRID:SCR_013597 | |
| Software and Algorithms | Thunder | Janelia | RRID: SCR_016556 | |
| Software and Algorithms | Spark | Apache | RRID: SCR_016557 | |
| Other | MIMMS microscope 1.0 (2016) | Janelia | RRID:SCR_016511 | |
| Other | Tip-Tilt-Z Sample Positioner | Janelia | RRID:SCR_016528 |
