Extended Data Figure 6: Input structures of CRF⁺ and SOM⁺ neurons, and axonal targets of CRF⁺ cells.

a, Pseudotyped-rabies tracing approach used to identify input areas to the CEA CRF⁺ or SOM⁺ population. A Cre-conditional AAV encoding the rabies glycoprotein and a G-deleted EnVA-pseudotyped rabies were injected into the CEA of mice. Mouse brain figure reproduced with permission from ref. 31. b, Left, merged image of glycoprotein and rabies expression localized to CRF⁺ cells of the CEA. Arrowheads indicate starter cells, which co-express rabies and glycoprotein. Inset, an example starter cell expressing both rabies and glycoprotein. Top right, rabies expression in the CEA. Bottom right, neurons expressing glycoprotein. c, Example regions providing monosynaptic input to the CRF⁺ network. vHIPP, ventral hippocampus; BLP, basolateral amygdaloid nucleus, posterior part; PVT, paraventricular thalamic nucleus. d, Comparison of the number of presynaptic cells to CRF⁺ and SOM⁺ cells, normalized to the total number of presynaptic cells in the defined regions. While there are a similar number of presynaptic neurons across brain regions projecting to SOM⁺ cells (Friedman test, P = 0.32), there are significantly different numbers of presynaptic neurons projecting to CRF⁺ cells (repeated-measures one-way ANOVA F = 14.39, P < 0.05). APir, amygdalopiriform transition area; BLA, basolateral amygdala; IC, insular cortex. e, Table listing all raw values from rabies-tracing experiments. f, Injection strategy used to locate terminal fields of CEA CRF⁺ axonal projections. A Cre-conditional AAV encoding synaptophysin conjugated to GFP was injected into the CEA of CRF-ires-cre mice. Mouse brain figure reproduced with permission from ref. 31. g, Expression of GFP in the CEA illustrating transfected somata and GFP⁺ puncta representing local axon terminals. h, CRF⁺ neurons project to multiple structures in the hypothalamus. PH, posterior hypothalamic area; LH, lateral hypothalamic area; PeF, perifornical nucleus; DM, dorsomedial hypothalamic nucleus; MTu, medial tuberal nucleus; VMH, ventromedial hypothalamic nucleus; Arc, arcuate hypothalamic nucleus. i, Demonstration of CRF⁺ terminals within the VMH. j, CEA CRF⁺ neurons also project to the peri-aqueductal grey (PAG). dmPAG, dorsomedial PAG; dl/lPAG, dorsolateral/lateral PAG; vlPAG, ventrolateral PAG. k, CEA CRF⁺ cells project widely throughout PAG columns.

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