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. 2021 Aug 23;12:5080. doi: 10.1038/s41467-021-25368-y

Fig. 3. The PVT-BNST projection provides polysynaptic inhibition onto BNSTCRF neurons.

Fig. 3

a–b Approach for slice recordings in BNSTCRF neurons during optical excitation of ChR2 in PVT axon terminals (a) and representative coronal image of the dorsal BNST (b); ac = anterior commissure, L = lateral, M = medial. Scale bar = 100 μM. Similar images were used to visually confirm ChR2 expression in each round of surgery and produced similar results. c–d Representative traces (c) and quantification (d) of time-locked optically-evoked EPSCs (oEPSCs) and IPSCs (oIPSCs) in BNSTCRF neurons in response to 2 ms pulses of blue LED at baseline, in the presence of tetrodotoxin (TTX, 1 μM), and with the addition of 4-aminopyridine (4-AP, 100 μM). oEPSCs: 1xRM-ANOVA effect of drug (F2,14 = 22.6, P < 0.0001), post hoc two-tailed paired t-tests with Holm-Sidak corrections (aCSF vs. TTX: t14 = 6.71, ****P < 0.0001; TTX vs. TTX + 4-AP: t14 = 3.16, **P = 0.007); N = 5 mice, 8 cells. oIPSCs: 1xRM-ANOVA effect of the drug (F2,10 = 429.0, P < 0.0001), post hoc two-tailed paired t-tests with Holm-Sidak corrections (aCSF vs. TTX: t10 = 25.90, ****P < 0.0001; TTX vs. TTX + 4-AP: t10 = 1.10, P = 0.299); N = 4 mice, 6 cells. e Paired pulse ratio of pharmacologically-isolated oEPSCs is lower in females than males (two-tailed unpaired t-test: t27 = 2.33, *P = 0.028) and below 1 in females (one-sample t-test: t12 = 4.07, ##P = 0.002) but not males (one-sample t-test: t15 = 2.33, P = 0.235). N’s = 4 M, 16 cells; 3 F, 13 cells. f oEPSC AMPA/NMDA ratios between sexes (two-tailed unpaired t-test: t22 = 1.61, P = 0.121). N’s = 4 M, 15 cells; 3 F, 9 cells. Data are presented as mean values ± SEM.