Fig. 1: Social buffering of fear: Experimental paradigm and quantification of freezing levels to conditioned stimuli “CS” during different sessions. | Nature Communications

Fig. 1: Social buffering of fear: Experimental paradigm and quantification of freezing levels to conditioned stimuli “CS” during different sessions.

From: Social buffering in rats reduces fear by oxytocin triggering sustained changes in central amygdala neuronal activity

Fig. 1: Social buffering of fear: Experimental paradigm and quantification of freezing levels to conditioned stimuli “CS” during different sessions.The alternative text for this image may have been generated using AI.

a Behavioral protocol. During habituation on Day 1, rats were exposed four times to two different auditory conditioning stimuli (CS1: 5 kHz, red; CS2: 15 kHz, blue) and subsequently fear conditioned by pairing each CS with an electric foot-shock of 0.5 mA (unconditioned stimulus; US). On Day 2 memory of fear was assessed by re-exposure to each CS (fear recall) and, 3 h later, fear memory was again assessed for 10 min during re-exposure to CS1 (as in (b) and further experiments) or CS2 (as in (c)) after introduction of a polyester ball (No Companion, upper panel) or a companion rat in the adjacent compartment (Companion, lower panel) to test the social buffering of the fear response to the CS (SBF). On Day 3, both CSs were presented again in a new cage without the companion (Retention of SBF). b Freezing levels are equally high upon recall in the “No companion” and “Companion” groups, but freezing to CS1 (5 kHz) is reduced by the presence of the companion (Day 2; “SBF”; F(3,20) = 19.47; p < 0.001), and one day later in the absence of the companion (Day 3; “Retention of SBF”, F(5,30) = 18.16; p < 0.001; ns, no effects without companion for CS1 and CS2). Freezing levels of rats across the experimental protocol as shown in (a) in the absence (“No companion”, filled bars, n = 6) or presence of a companion (“+Companion”, striped bars, n = 6). c Freezing to CS2 (15 kHz) was high upon recall in both groups, similar to (b), and reduced by the presence of the companion (Day 2) and again one day later (Day 3) revealing that both the acute (F(3,32) = 51.48, p < 0.001, ns, fear reduction without companion) and memory effects of SBF (F(5,48) = 19.97, p < 0.001; ns, no effects without companion for CS2 and CS1) are independent of CS frequency. (“No companion”, filled bars, n = 9; “+companion”, striped bars, n = 9). Pre-tone (white bars), basal freezing levels before testing. Two-way ANOVA (pre-tone, CS1, CS2) and group (No Companion, Companion), for each session (“Habituation”, “Fear recall”, “SBF”, “Retention of SBF”). All Bonferroni-corrected p-values are indicated in the figures. Individual and mean values ± SEM are shown. See also Supplementary Figs. 2–4.

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