Extended Data Fig. 7: Genetic targeting of osmotic and hypovolaemic thirst-activated cell populations in the SFO and the OVLT.

a, Spearman correlation between Fos expression under distinct thirst states and cell-type-specific and thirst-state-specific marker genes. Thirst-state-specific marker genes (Rxfp1 and Pdyn) show higher correlation with Fos expression compared to cell-type-specific genes. b, Two-colour in situ hybridization of Pdyn and Rxfp1. These gene expression patterns are mostly distinct with minor overlap (arrowhead). Representative images from 8 and 2 slices from 2 independent experiments for SFO (left) and OVLT (right) respectively. Scale bar, 10 μm. c, Validation of Cre expression in Pdyn-Cre and Rxfp1-Cre lines. 95.5% of Pdyn-Cre and 100% of Rxfp1-Cre expression matched endogenous gene expression. Representative images from 4 and 2 slices from 2 independent experiments for PDYN/CRE and RXFP1/CRE labelling, respectively. Scale bar, 10 μm. d, Immunostaining of the SFO (top) and OVLT (bottom). Shown are PDYN-positive neurons in Pdyn-Cre/Ai3 animals (representative images out of 8 slices from 4 mice for SFO and out of 2 slices from 2 mice for OVLT, left) and RXFP1-positive neurons in Rxfp1-Cre/Ai14 animals (representative images out of 6 slices from 3 mice for both SFO and OVLT, right). PDYN- and RXFP1-positive neurons (red) are a partial population of ETV1-positive excitatory neurons (green). Almost all (>90%) PDYN- and RXFP1-positive neurons expressed Etv1. RXFP1 and PDYN data are from Fig. 4d. Scale bar, 10 μm.