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. Author manuscript; available in PMC: 2017 Jul 21.
Published in final edited form as: Nat Neurosci. 2016 Sep 5;19(10):1356–1366. doi: 10.1038/nn.4377

Figure 7.

Figure 7

Th+ VTA neuron projections differentially modulate arousal. (a–d) Representative images of Th+ VTA fibers expressing ChR2 at the (a) NAc, (b) mPFC, (c) CeA and (d) DLS. Scale bars, 100 μm (one of six mice for a and b and one of four mice for c and d). Note that each column represents one output region. (e–h) Mean latency (± s.e.m.) to wake from NREM sleep following optogenetic stimulation at 1 and 25 Hz of eYFP and ChR2 mice (8–12 stimulations per frequency per mouse); two-way RM ANOVA between frequency and viral transduction (interactions: F1,10 = 5.4 (NAc), F1,12 = 0.94 (mPFC), F1,6 = 31.71 (CeA), F1,6 = 29.73 (DLS), P(NAc) = 0.042, P(mPFC) = 0.35, P(CeA) = 0.001, P(DLS) = 0.002), followed by Sidak’s post hoc tests (NAc: **P = 0.002, ****P = 9 × 10−6, CeA: **P = 0.001, ****P = 1 × 10−5, DLS: *P = 0.046, ***P = 2 × 10−4, ****P = 4 × 10−7). (i–l) Mean atency (± s.e.m.) to wake from REM sleep following optogenetic stimulation at 1 and 25 Hz of eYFP and ChR2 mice (8–12 stimulations per frequency per mouse); two-way RM ANOVA between frequency and viral transduction (interactions: F1,10 = 0.8 (NAc), F1,12 = 5.081 (mPFC), F1,6 = 1.032 (CeA), F1,6 = 1.94 (DLS); P(NAc) = 0.3, P(mPFC) = 0.043, P(CeA) = 0.35 , P(DLS) = 0.2; virus (CeA): F1,6 = 6.207, P = 0.047) followed by Sidak’s post hoc tests (mPFC: **P = 0.003, ****P = 9 × 10−6). (m–x) The percentage (mean ± s.e.m.) of time spent (m–p) awake, (q–t) in NREM sleep and (u–x) in REM sleep during 24 h starting with photostimulation of the different output regions in eYFP (white) and ChR2 (blue) mice. n = 6 per group for NAc and mPFC and n = 4 per group for CeA and DLS experiments; two-way RM ANOVA between time and viral transduction (NAc interactions: wake F23,115 = 2.829, P = 0.0001; NREM F23,115 = 2.455, P = 0.0009; REM: F23,115 = 2.938, P = 8 × 10−5), followed by Sidak’s post hoc tests (NAc wake: P(ZT 0) = 0.005, P(ZT 1) = 0.022, P(ZT 2) = 0.0004; NREM: P(ZT 0) = 0.003, P(ZT 2) = 0.001; REM: P(ZT 1) = 0.01, P(ZT 2) = 0.005).