Supplementary Figure 1: PVN CRFR1⁺ neurons represent a distinct population of neurons residing in the PVN.

(a). PVN-CRFR1⁺ neurons are significantly smaller in size than OT⁺ and AVP⁺ neurons. Volumetric measurements revealed significant differences in somatal-volume between PVN-CRFR1⁺ neurons and OT⁺ and AVP⁺ neurons (One-way ANOVA, F_(3,521)=84.3, p<0.001 followed by Tukey’s post-hoc analysis). Data presented as mean ± SEM, n=5 mice. (b) Brain coronal sections obtained from CRF^Cre x CRFR1^GFP mice following injection of DIO-mCherry virus reveal PVN-CRF⁺ fibers adjacent to PVN-CRFR1-GFP⁺ neurons. Magnification at 40X. Scale bar represents 10 μm (c) Representative image showing single-cell reverse-transcription-PCR products of 5 PVN neurons positive for CRFR1-GFP. Amplification of Gapdh mRNA served as an internal positive control and data was only included when there was Gapdh positive signal. Eleven out of 12 PVN-CRFR1⁺ neurons express mRNA for Gad1 or Gad2, the GABA synthesizing enzymes.