Figure 2. PRF preferentially activates subregions of mPFC and BNST.

(A) Schematic depiction of experimental procedure for assessing ex vivo neuronal regional activity (via c-Fos immunohistochemistry) after PRF or FRF retrieval, along with CS-only controls. Representative images and c-Fos+ cell count differences for basal amygdala (B), paraventricular nucleus of the thalamus (C), infralimbic cortex (D), prelimbic cortex (E), posterior portion of the anterior cingulate cortex (F), anteroventral BNST (G), anterodorsal BNST (H), and lateral habenula (I). For corresponding behavioral data, see Figure 1B. Scale bars = 30 µm (B,D–F), 100 µm (C,I), 300 µm (G,H). n = 4–8 mice per group. Data are means ± SEM. *p<0.05.

Figure 2—figure supplement 1. Ex vivo neuronal regional activity correlates of PRF.

(A) Schematic depiction of experimental procedure for assessing ex vivo neuronal regional activity (via c-Fos immunohistochemistry) after PRF or FRF retrieval. Representative images and c-Fos+ cell counts for the dorsal dentate gyrus (B), dorsal CA3 (C), and ventral CA1/subiculum (D) regions of the hippocampus and the medial habenula (E). There were no group differences. Scale bars = 100 µm. n = 4–8 mice per group. Data are means ± SEM.

Figure 2—figure supplement 2. Connectivity between mPFC, BNST, and downstream targets.

(A) Schematic depiction of viral strategy to label mPFC inputs to BNST neurons and their onward projections to the hypothalamus. (B) Representative image of synaptophysin-mCherry labeling in BNST neurons receiving mPFC input (scale bar = 1000 µm). (C) Representative image of synaptophysin-mCherry labeling in mPFC-innervated BNST neuronal projections in hypothalamus (scale bar = 500 µm). (D) Schematic depiction of viral strategy to label PVN oxytocin cell inputs to the PAG in Oxt-Cre mice. (E) Representative image of ChR2-EYFP labeling in the PVN (scale bar = 200 µm). Low (F) and high (G) magnification images of ChR2-EYFP labeling in the PAG (scale bars = 200 µm and 100 µm, respectively). (H) Schematic depiction of viral strategy to label PVN oxytocin cell inputs to the PAG in Oxt-Cre mice. (I) Representative image of synaptophysin-EYFP labeling in the PVN (scale bar = 200 µm). Low (J) and high (K) magnification images of synaptophysin-EYFP labeling in the PAG (scale bars = 200 µm and 100 µm, respectively). Note: LH = Lateral Hypothalamus; PVN = paraventricular nucleus of the hypothalamus; IPAG = lateral periaqueductal gray; vlPAG = ventrolateral periaqueductal gray; Aq = aqueduct.