Prefrontal Cortex Drives Distinct Projection Neurons in the Basolateral Amygdala  Laura M. McGarry, Adam G. Carter  Cell Reports  Volume 21, Issue 6, Pages 1426-1433 (November 2017) DOI: 10.1016/j.celrep.2017.10.046 Copyright © 2017 The Authors Terms and Conditions

Cell Reports 2017 21, 1426-1433DOI: (10.1016/j.celrep.2017.10.046) Copyright © 2017 The Authors Terms and Conditions

Figure 1 PFC Axons Overlap with Multiple Projection Neurons in the BLA (A) (Left) Injections into the PL PFC to label axons and neurons in the BLA. Axes apply to all images. (Right) Injection of AAV-mCherry (red) and CTB-488 (green) in the PL PFC (top left) labels PFC axons (red) and AC neurons (green) in the BLA (bottom middle), with an expanded view (far right). DAPI staining in gray. (B) Average PFC-evoked AMPAR EPSCs at AC neurons before (green) and after (black) wash-in of NBQX. Arrowhead indicates time of light pulse. (C) (Left) Dual injections of CTB into the PFC and NAc. (Right) Labeling of AC (green) and AS (purple) neurons in the BLA. (D) (Left) Dual injections of CTB into the PFC and vHPC. (Right) Labeling of AC (green) and AH (orange) neurons in the BLA. (E) (Left) Average PFC-evoked AMPAR EPSCs at pairs of AS neurons located in the medial (purple) and lateral (black) BLA. (Right) Summary of EPSC amplitudes at AS neurons located in medial (M) or lateral (L) BLA. (F) (Left) Average PFC-evoked AMPAR EPSCs at AH neurons located in the medial (orange) and lateral (black) BLA. (Right) Summary of EPSC amplitudes at AH neurons located in medial (M) or lateral (L) BLA. Cell Reports 2017 21, 1426-1433DOI: (10.1016/j.celrep.2017.10.046) Copyright © 2017 The Authors Terms and Conditions

Figure 2 PFC Inputs Are Strongest in AC and AH Neurons (A) Dendritic reconstructions of AC (green), AS (purple), and AH (orange) neurons located in the medial BLA. (B) Summary of dendritic Sholl analysis for the five types of neurons. (C) Average PFC-evoked AMPAR and NMDAR EPSCs at pairs of neighboring AC (green) and AS (purple) neurons. (D) Summary of AS/AC amplitude ratio (left) and AMPA/NMDA ratio (right) for PFC-evoked EPSCs at AC and AS neurons. (E and F) Similar to (C) and (D) for AC (green) and AH (orange) neurons. ∗p < 0.05. Cell Reports 2017 21, 1426-1433DOI: (10.1016/j.celrep.2017.10.046) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Differential Synaptic Strength Reflects Number of Connections (A) (Top) PFC-evoked qEPSCs recorded in the presences of strontium (Sr2+) at AS (purple), AC (green), and AH (orange) neurons, where asterisks indicate detected qEPSCs. Arrowheads indicate time of light pulses. (Bottom) Histograms for example cells showing detected post-stimulation qEPSCs across all trials. (B) Summary of PFC-evoked qEPSC Δ frequency (left) and amplitude (right) at AS, AC, and AH neurons. Lines indicate pairs of recorded neurons. (C) Summary of AS/AC (left) and AH/AC (right) ratios for PFC-evoked qEPSC Δ frequency and amplitude. (D) (Left) Average PFC-evoked AMPAR EPSCs evoked by trains of inputs (5 pulses at 20 Hz) at AC (green) and AS (purple) neurons. (Right) Summary of paired-pulse ratio (PPR = EPSCN / EPSC1) during trains. (E) Similar to (D) for AC (green) and AH (orange) neurons. ∗p < 0.05. Cell Reports 2017 21, 1426-1433DOI: (10.1016/j.celrep.2017.10.046) Copyright © 2017 The Authors Terms and Conditions

Figure 4 PFC Inputs Preferentially Drive Firing of AC and AH Neurons (A) AP firing and hyperpolarization of AC (green), AS (purple), and AH (orange) neurons located in the medial BLA in response to +200 pA and −50 pA current steps in the presence of synaptic blockers. (B) Firing versus current (F-I) curves for the five types of neurons. (C) (Left) PFC-evoked EPSPs and APs at neighboring AC (green) and AS (purple) neurons. Example traces to light duration of 4 ms shown. Arrowhead indicates time of light pulse. (Right) Probability of AP firing versus light duration. (D) Similar to (C) for AC (green) and AH (orange) neurons. (E) (Left) PFC-evoked EPSPs and APs at neighboring AC (green) and AS (purple) neurons in response to trains (5 pulses at 20 Hz). Example traces to 4 ms light duration shown. (Right) Probability of AP firing versus pulse number. (F) Similar to (E) for AC (green) and AH (orange) neurons. ∗p < 0.05. Cell Reports 2017 21, 1426-1433DOI: (10.1016/j.celrep.2017.10.046) Copyright © 2017 The Authors Terms and Conditions