Figure 3. Changes sleep-wake following Nts^LH neuron lesions.

(A) Representative brain section from Nts-GFP mouse injected with AAV-DTA into the lateral hypothalamus. Nts+ neurons were visualized by native GFP fluorescence (Green cells). AAV-DTA selectively kills Cre+ Nts neurons but expresses mCherry in the Cre-negative neurons (Red cells), thereby marking the injection site. Note the absence of GFP+ neurons within the boundaries of injection site. Percentages of wake, REM and NREM sleep during light (C) dark (D) or the entire 24-hr period (B) in mice with complete (>75% bilateral cell loss; black bars) or incomplete Nts^LH lesions (gray bars) and sham controls (white bars). (B)(D) *P < 0.05: One-way ANOVA followed by Tukey’s multiple comparisons test (for B, wake: n =7–8: F = 4.88, P = 0.019, control vs complete lesion: P = 0.015, NREM: n =7–8: F = 4.12, P = 0.032, control vs complete lesion: P = 0.025) (for D, wake: n =7–8: F = 5.41, P = 0.013, control vs complete lesion: P = 0.010, NREM: n =7–8: F = 5.33, P = 0.014, control vs. complete lesion: P = 0.010). Fragmentation index of sleep-wake stages (number of bouts as a function of time spent in that stage normalized to control mice) are shown E, indicating selective fragmentation of wake in mice with complete Nts^LH lesions. *P < 0.05, **P < 0.01: One-way ANOVA followed by Tukey’s multiple comparisons test (n = 7–8: F = 8.29, P = 0.0031, wake vs NREM: P = 0.0052, wake vs REM: P = 0.011). Values are the mean ± SEM.