Fig. 6.
Optogenetic activation of the dorsal–intermediate lateral septum (LS) γ-aminobutyric acid–mediated (GABAergic) neuronal terminals in the ventral tegmental area (VTA) promoted wakefulness and shortened emergence time from general anesthesia. (A) Schemetic showing the virus injection into the dorsal–intermediate lateral septum and optical fiber implantation into the ventral tegmental area. (B) Typical image showing the GCaMP6s-labeled neuronal terminals in the ventral tegmental area and the location of the optical fiber. Scale bar, 100 μm. (C) Representative traces of electroencephalogram (EEG)/electromyography (EMG) recordings and EEG spectrum analysis showing an immediate sleep-to-wakefulness transition during non–rapid eye movement (non-REM) sleep after acute photostimulation (20 Hz for 30 s) of the dorsal–intermediate lateral septum GABAergic terminal in the ventral tegmental area. (D) Summary showing that compared to the enhanced yellow fluorescent protein (eYFP) mice (gray), the channelrhodopsin-2 (ChR2) mice (blue) exhibited lower latencies to wake from non-REM sleep after optogenetic stimulation at 20 Hz. U = 0.00, P < 0.001, Mann–Whitney rank sum test (two-tailed). (E through H) Statistical results for power distribution of EEG frequency bands of delta, theta, beta, and gamma during optogenetic activation of dorsal–intermediate lateral septum GABAergic terminals in the ventral tegmental area. Unpaired two-tailed t test, delta: t10 = 5.02, P < 0.001; theta: t10 = 0.61, P = 0.553; Mann–Whitney rank sum test (two-tailed), beta: U = 6.00, P = 0.050; gamma: U = 8.00, P = 0.109. (I) Left: Schematic of sustained optical stimulation protocol after the discontinuation of isoflurane administration. Right: Summary data showing that activation of the dorsal–intermediate lateral septum GABAergic neuronal terminals in the ventral tegmental area accelerated emergence time from isoflurane anesthesia. t10 = 10.21, P < 0.001, unpaired two-tailed t test. The data represent means ± SD. *P < 0.05; ***P < 0.001. DAPI, 4′,6-diamidino-2-phenylindole.

Optogenetic activation of the dorsal–intermediate lateral septum (LS) γ-aminobutyric acid–mediated (GABAergic) neuronal terminals in the ventral tegmental area (VTA) promoted wakefulness and shortened emergence time from general anesthesia. (A) Schemetic showing the virus injection into the dorsal–intermediate lateral septum and optical fiber implantation into the ventral tegmental area. (B) Typical image showing the GCaMP6s-labeled neuronal terminals in the ventral tegmental area and the location of the optical fiber. Scale bar, 100 μm. (C) Representative traces of electroencephalogram (EEG)/electromyography (EMG) recordings and EEG spectrum analysis showing an immediate sleep-to-wakefulness transition during non–rapid eye movement (non-REM) sleep after acute photostimulation (20 Hz for 30 s) of the dorsal–intermediate lateral septum GABAergic terminal in the ventral tegmental area. (D) Summary showing that compared to the enhanced yellow fluorescent protein (eYFP) mice (gray), the channelrhodopsin-2 (ChR2) mice (blue) exhibited lower latencies to wake from non-REM sleep after optogenetic stimulation at 20 Hz. U = 0.00, P < 0.001, Mann–Whitney rank sum test (two-tailed). (E through H) Statistical results for power distribution of EEG frequency bands of delta, theta, beta, and gamma during optogenetic activation of dorsal–intermediate lateral septum GABAergic terminals in the ventral tegmental area. Unpaired two-tailed t test, delta: t10 = 5.02, P < 0.001; theta: t10 = 0.61, P = 0.553; Mann–Whitney rank sum test (two-tailed), beta: U = 6.00, P = 0.050; gamma: U = 8.00, P = 0.109. (I) Left: Schematic of sustained optical stimulation protocol after the discontinuation of isoflurane administration. Right: Summary data showing that activation of the dorsal–intermediate lateral septum GABAergic neuronal terminals in the ventral tegmental area accelerated emergence time from isoflurane anesthesia. t10 = 10.21, P < 0.001, unpaired two-tailed t test. The data represent means ± SD. *P < 0.05; ***P < 0.001. DAPI, 4′,6-diamidino-2-phenylindole.

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