Fig. 1: CRHR1 mRNA is highly expressed in parvalbumin-positive TRN neurons.

a Example image overview of RNAscope in situ mRNA hybridization stained for DAPI (blue), with parvalbumin (PV—magenta) highlighting the TRN and white squares indicating the corresponding regions shown as close-ups in hippocampal CA3 (b), TRN (c), and basolateral amygdala (BLA, d), where CRHR1 mRNA can be seen as white puncta. Quantification from independent observations is provided in (e). e TRN has significantly higher levels of CRHR1 mRNA compared to CA3 and BLA (n = 11). f No significant difference was found between males (n = 6) and females (n = 5) in CRHR1 mRNA expression in TRN. g Example image of in situ mRNA hybridization for parvalbumin (PV—magenta), somatostatin (SOM—green), and CRHR1 (white) mRNA labeling in TRN, showing the expected anatomical segregation into a PV+ core and SOM+ shell in the sensory sector. h Close-up of TRN, exemplifying a higher density of CRHR1 mRNA puncta in PV+ neurons compared to SOM+ neurons. i CRHR1 mRNA has a significantly higher expression in PV+ TRN neurons. Each point is the average density of CRHR1 mRNA in PV+ or SOM+ only TRN nuclei detected with DAPI of a mouse (n = 11). Data are represented as mean ± SEM. All scale bars are 100 µm. Statistical analysis was performed by e repeated measures one-way ANOVA with Holm–Šídák’s multiple comparisons test and f–i two-tailed unpaired t-test, with **p < 0.01, ***p < 0.001. For additional statistical information, see Supplementary Table 1. Source data are provided as a Source data file.