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DOI: 10.3791/4328-v
This procedure outlines the manipulation of cerebral cortical pyramidal neurons using optogenetics while monitoring electroencephalogram (EEG), electromyogram (EMG), and cerebral lactate concentration. The experimental setup is conducted on cable-tethered mice during spontaneous sleep/wake cycles.
A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.
The overall aim of this procedure is to measure EEG defined sleep and lactate concentration during optogenetic stimulation at precise stereotaxic coordinates. Surgically implant the necessary electroencephalographic leads and cannulas for a lactate sensor and fiber optic cable. Then insert the fiber optic cable and pre calibrated lactate sensor into their guide cannulas in the animal's skull.
Next, adjust the blue light stimulus intensity to achieve the desired electrophysiological response. Then proceed to collect the electroencephalographic, electromyographic and lactate concentration data while the animal is allowed to spontaneously behave and or sleep. Compared to other methods like pharmacological studies of the EEG, this experimental approach has the advantage of monitoring brain biochemistry and electrophysiology while the activity of neurons is manipulated in real time on a sub-second time scale.
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