Fig. 2: Chemogenetic activation of PSTN CR neurons increased wakefulness and exploratory behaviors. | Nature Communications

Fig. 2: Chemogenetic activation of PSTN CR neurons increased wakefulness and exploratory behaviors.

From: Parasubthalamic calretinin neurons modulate wakefulness associated with exploration in male mice

Fig. 2: Chemogenetic activation of PSTN CR neurons increased wakefulness and exploratory behaviors.The alternative text for this image may have been generated using AI.

a Diagram of the bilateral viral infection area of AAV-hSyn-DIO-hM3Dq-mCherry in the PSTN. b Representative photomicrographs of the PSTN depicting mCherry (red), c-Fos (green), and merge (yellow) images of PSTN-mCherry and PSTN-hM3Dq mice after administration of CNO. PSTN: parasubthalamic nucleus, cp: cerebral peduncle. Scale bar: 50 µm. c Representative voltage traces recorded from an mCherry-expressing neuron and an hM3Dq-expressing neuron during the application of CNO, which produced depolarization and firing in a patched hM3Dq-expressing neuron. d Examples of relative EEG power, EEG/EMG traces, and hypnograms over 6 h following CNO injection of PSTN-mCherry or PSTN-hM3Dq mice at 09:00. e Time course changes in wakefulness, NREM sleep, and REM sleep after administration of CNO to PSTN-mCherry or PSTN-hM3Dq mice (mCherry mice: n = 6, hM3Dq mice: n = 8; Two-way ANOVA, Wake: F1,12 = 76.13, P = 1.53 × 10−6; NREM: F1,12 = 78.02, P = 1.34 × 10−6; REM: F1,12 = 8.66, P = 0.0123). f Total time spent in each stage for 4 h after CNO injection of PSTN-mCherry or PSTN-hM3Dq mice. (mCherry mice: n = 6, hM3Dq mice: n = 8; unpaired two-sided t-test, Wake: t12 = 17.76, P = 5.548 × 10−10; NREM: t12 = 16.69, P = 1.142 × 10−9 REM: t12 = 14.8, P = 4.542 × 10−9). g EEG power density of wakefulness, NREM sleep, and REM sleep during the 4 h after CNO injection of PSTN-mCherry or PSTN-hM3Dq mice (mCherry mice: n = 6, hM3Dq mice: n = 8; Two-way ANOVA; not statistically significant). h Quantification of the behaviors observed in mice in the 1 h following CNO or saline injection at 21:00 (n = 8, paired two-sided t-test; sniff: t7 = 3.041, P = 0.0188; rear: t7 = 2.448, P = 0.0443). Bars represent the mean percentage of awaken time (±SEM) that mice spent conducting each behavior. i, j Chemogenetic activation of PSTN CR neurons increased the locomotion and center time in the open-field test (n = 8, unpaired two-sided t-test; total distance: t7 = 4.806, P = 0.0003; center time: t7 = 2.502, P = 0.0254) (i) and the number of entries in the white box (mCherry groups and hM3Dq+vehicle: n = 7, hM3Dq+CNO: n = 8; unpaired two-sided t-test, t13 = 3.604, P = 0.0032) (j). Data represent mean ± SEM, *P < 0.05, **P < 0.01.

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