Fig. 7: CRH modulates NREMS through activation of CRHR1 in PV+ TRN neurons.

a Representative image showing the expression of the shRNAmir virus used to knock down (KD) CRHR1 in TRN. The GFP reporter for shRNAmir expression (green) is colocalized with parvalbumin (PV—magenta) in TRN neurons (scale bars are 200 µm). Comparable expression was found in n = 6 infected mice. b Example images (infrared and fluorescence channel) of patched-clamped TRN neurons from mice injected with a control GFP or shRNAmir (scale bars are 10 µm). Example traces from current-clamp recordings showing that CRHR1 KD prevented the changes in TRN bursting induced by bath application of CRH (500 nM). c Quantification of the CRH-induced decrease in TRN bursting, which was abolished by the CRHR1 KD. d Example images for the GFP injected controls, left—optic fiber implantation over the sensory TRN in CRH-IRES-Cre × Ai27D mice implanted for EEG/EMG recordings and right—channelrhodopsin2 (ChR2—red) expression in CRH-positive axons and GFP (green) labeled TRN neurons. Scale bars are 200 µm. e In control mice injected with GFP, photoactivation of CRH release during NREMS increased the number of MAs, decreased the average sigma power (f), and prolonged the latency to REMS from the sigma peak (g). h Example images for the shRNAmir injected mice, left—optic fiber implantation over the sensory TRN in CRH-IRES-Cre × Ai27D mice implanted for EEG/EMG recordings and right—ChR2 (red) expression in CRH-positive axons and the shRNAmir expression labeled with GFP (green) in TRN PV+ neurons. Scale bars are 200 µm. i–k CRHR1 KD abolished the NREMS fragmentation induced by photoactivation of CRH release. Data are represented as mean ± SEM. Statistical analysis was performed by (b) one-tailed unpaired t-test and (e–g, i–k) one-tailed paired t-test with *p < 0.05, **p < 0.01, ***p < 0.001. For additional statistical information, see Supplementary Table 1. Source data are provided as a Source data file.