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. Author manuscript; available in PMC: 2017 Nov 17.
Published in final edited form as: Nature. 2017 May 17;545(7655):477–481. doi: 10.1038/nature22350

Extended Data Figure 4. Optogenetic manipulation of axon projections of POA GABAergic neurons to TMN, DMH, and Hb and effect of anti-histamine on optogenetic activation of the TMN axon projections.

Extended Data Figure 4

a, AAV-DIO-ChR2-eYFP or AAV-DIO-iC++-eYFP was injected into the POA of GAD2-Cre mice and an optic fiber was implanted into the TMN for optogenetic activation/inhibition. Mouse brain figure adapted with permission from ref. 31. b, Percentage of time in wake, NREM, or REM state before, during, and after laser stimulation (blue shading, 10 Hz, 120 s) in mice expressing ChR2, averaged from 9 mice (P<0.0001 for wake, REM and NREM, bootstrap). Shading for each trace, 95% confidence interval (CI). c, Similar to b, but in mice expressing iC++ (blue shading: constant light, 60 s), averaged from 4 mice (P<0.0001 for wake and NREM, P=0.004 for REM, bootstrap). d, Schematic for optogenetic activation of POA GABAergic projection to the habenula (Hb). e, Similar to b, for activating POA→Hb projection (P=0.28, 0.35 and 0.72 for REM, wake and NREM, bootstrap, n=3 mice). f, Schematic for optogenetic activation of POA GABAergic projection to the dorsomedial hypothalamus (DMH). g, Similar to b, for POA→DMH activation (P=0.02, 0.12, and 0.09 for wake, REM, and NREM, n=3 mice). h, Left, schematic for optogenetic activation of POA GABAergic projection to the TMN without drug treatment. Right, percentage of time in wake, NREM, or REM state before, during, and after laser stimulation (blue shading, 10 Hz, 120 s) in mice with no drug (effect of laser: P<0.001 for wake, REM and NREM, bootstrap; n=14 mice). i, Similar to h, but after injection of triprolidine (20 mg/kg, i.p.; effect of laser: P=0.21, 0.84, 0.57 for wake, REM and NREM, n=5 mice). j, Percentage of time in NREM, REM or wake state before and during laser stimulation in no drug and triprolidine groups (120 s periods before and during laser stimulation, *P<0.05, **P<0.01, ***P<0.001; ns, P>0.05, signed rank test between before and during laser, rank sum test between no drug and triprolidine for the period before laser stimulation). k, Laser-induced change in the percentage of each state (difference between the 120s periods before and during laser stimulation, *P<0.05, rank sum test). Error bar, ±s.e.m.