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. Author manuscript; available in PMC: 2017 Nov 12.
Published in final edited form as: Neuroscience. 2016 Sep 3;336:102–113. doi: 10.1016/j.neuroscience.2016.08.046

Figure 1. Specific expression of AAV-hM3Dq-mCherry in MCH neurons in MCH-Cre mice and their activation by CNO.

Figure 1

A. Photomicrograph of a brain section labeled for mCherry (red cells) and MCH (green cells) by immunofluorescence from a mouse injected with AAV-hM3Dq into the lateral hypothalamus indicating DREADD expression in the MCH neurons (white arrows indicate doubly labeled neurons). The percentage of MCH neurons transfected ranged from 21–60%. B- Representative whole cell, current clamp recording from a MCH neuron (identified by mCherry fluorescence) indicating bath application of CNO (500 nM) induced depolarization and increased the firing rate of these neurons. Intraperitoneal (IP) injections of CNO induced cFos expression (black nuclei) in DREADD-expressing neurons (brown cytoplasm) in the lateral hypothalamus (C). Double labelling of sections for MCH (brown) and cFos (black) indicated cFos expression in MCH neurons after IP CNO (black arrows in D). Brains were obtained for cFos analysis 180 minutes after IP CNO (0.3 mg/kg of body weight). Activation of MCH neurons by IP CNO caused specific increases in REM sleep in mice. REM sleep amounts (in minutes) during the first two 4 hr periods following IP saline or CNO injections either during the light (E; n=11) or dark period (F; n=9) in MCH-Cre mice injected with AAV-hM3Dq into the LH. Data are mean ± SEM. **P<0.01; ***P<0.001; paired t test.