Dual-label fluorescent immunohistochemistry comparing the adult (P60) female (a-d) and male (e-h) AVPV/PeN CRFR1-GFP, tyrosine hydroxylase (TH), and co-localization of CRFR1/TH (d, h; high-magnification display of images c, g respectively). (i) Comparison of female versus male CRFR1-GFP- (green), TH-expressing cells (magenta), and co-expression of TH in CRFR1-GFP cells (white) illustrating significant sex differences in P60 CRFR1-GFP-ir, TH-ir, and co-labeled cells (all F>M). (j) Female versus male comparison of the percentage of CRFR1 cells that co-localize TH, and percentage of TH cells that co-express CRFR1 (both F>M). * indicates statistical significance (p≤0.05), data are reported as mean ± SEM. Inset boxes indicate regions that were further magnified. Arrows indicate examples of co-localized cells. N=5 per sex. 3V, third ventricle.