Fig. 1: Characterization of the PB-CeC/L pathway and optogenetic LTP induction protocols. | Communications Biology

Fig. 1: Characterization of the PB-CeC/L pathway and optogenetic LTP induction protocols.

From: Aversive experiences induce valence plasticity of instructive signals to change future learning rules in mice

Fig. 1

a Schematic of AAV injection and electrophysiological analysis using acute brain slices containing the central amygdala. PB, lateral parabrachial nucleus. b Image showing Chronos:GFP expression in the proximity of the CeC/L region. Scale bars represent 200 μm (left panel) and 20 μm (right panel). BLA basolateral amygdala; Rec, recording pipette. c Traces of EPSCs (gray, 15 consecutive responses; red, average) evoked by photo-stimulation (every 20 s, 5 ms duration). d Time course of EPSC amplitude. e Summary of EPSC amplitude. Data are represented as mean + SEM. f Normalized leEPSC amplitude. The blue bar represents optogenetic high-frequency stimulation (oHFS) after a 10 min baseline recording. An oHFS consisting of five trains (0.1 Hz, each train at 20 Hz [n = 5 cells], 40 Hz [n = 14 cells], or 50 Hz [n = 8 cells], 0.5 s) was applied. g Representative traces of the leEPSCs at the indicated time points. h Plots of mean leEPSC potentiation 25–30 min after oHFS application. i Behavioral and electrophysiological scheme. The day after in vivo optogenetic LTP induction, ex vivo verification was performed using an electrophysiological approach. j Input-output relationship at PB-CeC/L synapse. Representative traces of leEPSCs (average of eight consecutive traces) recorded with increasing stimulus intensities in the oHFS and no-stimulated mice and summary of input-output relationship (No-stim; n = 18 cells, oHFS; n = 18 cells). k Summary of EPSC amplitude evoked by stimulation with 0.125 mW/mm2. l The NMDA/AMPA ratio at PB-CeC/L synapse. Representative traces of leEPSCs (average of 15 consecutive traces) recorded at holding potentials of −60 mV and +40 mV in the oHFS and no-stimulated mice and summary of the NMDA/AMPA ratio (No-stim; n = 8 cells, oHFS; n = 14 cells). m, n Ex vivo verification after successive threat conditioning. Representative average traces (upper) and scaled overlay traces (lower) of leEPSCs (average of 15 consecutive traces) in the oHFS and no-stimulated mice, and summary of the leEPSC amplitude (m) and PPR (n) (No-stim; n = 16 cells, oHFS; n = 22 cells). e One-way repeated-measures ANOVA and Dunnett’s multiple comparisons test. h One-sample t-test. k, l: Two-tailed unpaired t-test. m, n Mann–Whitney test. ns, p > 0.05; *p < 0.05, **p < 0.01.

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