E-Scope: an integrated device allowing synchronous calcium imaging of anterior cingulate cortex and electrophysiological recordings in cerebellum.

(A) Photograph of E-Scope hardware. The multichannel silicon probe (32 channels) connects to the custom Ephys PCB. The Ephys PCB is connected to the CMOS sensor PCB of the Miniscope via a 6-pin connector. The electrophysiology and image data streams are both conveyed through the coaxial cable. (B) Illustration of the process for implanting the E-Scope. (C) Illustrations and photomicrographs showing the location of AAV1-Syn-GCaMP6f virus injection in ACC (left, middle) and multichannel probe implant in the dentate nucleus of the cerebellum (left, right). (D) Pseudo-color (top left) and averaged activity heatmap from calcium imaging ACC neurons segmented using CNMF-E (bottom left). Calcium signals of ACC neurons (right). (E) In vivo single unit electrophysiology recording of PCs in the cerebellum (Cb). (F) Average spike waveform of a dentate nucleus (DN) neuron. (G) Average simple spike (SS) waveform (left) and average complex spike (CS) waveform (middle) of a PC. Cross-correlogram of simple spikes and complex spikes shows the pause in simple spike activity after a complex spike (right).

Purkinje cell and dentate nucleus neuron activity patterns during social behavior.

(A) Graph of the number of interaction bouts between the recorded subject mouse and a novel target mouse or object (two-sided Wilcoxon rank sum test; p = 0.0245, Z = 2.2485) (B) Illustration of probe location in the PC layer (top). Average simple spike waveform of a PC (bottom). (C) Heatmap of normalized (Z-score) firing rates of a Soc– PC neuron aligned to the onset of social interaction shown for 10 interaction bouts (top). The mean normalized firing rate across all interaction sessions shown above (bottom). (D, E, F) Average activity of 3 Soc+ PCs (D), 10 Soc– PCs (E), and 17 ns PCs (F). The mean activity of each group is shown below each heat map. (G) Proportion of Soc+, Soc– and ns PCs in the recorded population. (H) Illustration showing the path of the recorded animal in the social interaction arena (480 × 480 mm). (I) Illustration of probe location for DN recordings (top). Average spike waveform of a DN neuron (bottom). (J) Normalized firing rates of a Soc+ DN neuron aligned to the onset of social interaction shown for 18 interaction bouts (top). The mean normalized firing rate across all interaction sessions shown above. (K, L, M) Average activity of 19 Soc+ DN neurons (K), 10 Soc– DN neurons (E), and 63 ns DN neurons (F). The mean activity of each group is shown below each heat map. (N) Proportion of Soc+, Soc– and ns DNs in the recorded population.

ACC neuron activity patterns during social behavior.

(A) Illustration of GRIN lens implant location (top left). Raw image from ACC calcium imaging recording (middle left). Segmented and averaged activity heatmap from the recording shown above (bottom left). Example of raw behavioral trace of subject mouse (right). (B, C) Heatmap depicting the calcium activity of 10 Soc+ ACC neurons (B) and 15 Soc– ACC neurons (C) during a single social interaction session. Social interaction bouts are shown as gray bars. The average calcium activity is shown below each heatmap. (D, E) Top: Z-scored Soc+ (D) and Soc– (E) ACC neurons calcium activity for all neurons during all social interaction sessions. The onset of social interaction is marked as a dashed black line. Bottom: Mean of Z-scored activity shown for the heatmaps above for Soc+ (D) and Soc– (E) neurons. (F) Percentage of units showing significant modulation by social interaction. (G) Overlap of Soc+ and Obj+ ACC neuronal populations. (H) Overlap of Soc– and Obj– ACC neuronal populations.

Correlated activity in the cerebellar-cortical circuit during social behavior.

(A) Illustration of the cerebellar-cortical circuit. PCs provide converging inhibition to DNs that excite thalamic neurons. Thalamic neurons excite ACC neurons. (B) Simultaneously recorded calcium traces from 9 ACC neurons and the electrophysiologically recorded firing rate of a single PC (red) during a social interaction bout. Social interaction bouts are shown as gray bars. (C) Simultaneously recorded calcium traces from 10 ACC neurons (blue) and the electrophysiologically recorded firing rate of a single DN neuron during a social interaction bout. Social interaction bouts are shown as gray bars. (D-G) Cumulative histogram of the distribution of the correlation coefficients for the activity of (D) Soc+ PCs, (E) Soc– PCs, (F) Soc+ DNs, or (G) Soc– DNs with Soc+ (dark blue), Soc– (light blue), and Socns (blue gray) ACCs. Insets: cumulative histogram of the activity of each set of neurons calculated during periods when the mouse was not engaged in social interaction. (H, I, J, K) Correlation matrix showing percentage of cell pairs showing significant positive (H, J) or negative (I, K) correlations in activity between Soc+ and Soc– PCs (H, I) or Soc+ and Soc–DNs (J, K) with Soc+, Soc– and Socns ACC neurons. The color of the squares represents the proportion of neurons correlated.