Skip to main content
. Author manuscript; available in PMC: 2019 Jul 3.
Published in final edited form as: Cell Metab. 2018 May 31;28(1):55–68.e7. doi: 10.1016/j.cmet.2018.05.001

Figure 5: Viral transduction of Designer Receptors Exclusively Activated by Designer Drugs (DREADDs; hM3Gq) is a valid method of inducing selective and functionally-relevant activation of MCH neurons.

Figure 5:

(A) Representative images of the mCherry reporter gene present in MCH neurons, indicating successful infection of the DREADDS AAV selective in MCH neurons. MCH: Green; mCherry: Red; Colocalization: Yellow. Scale bar: 50 μm. (B) Representative differential interference contrast (DIC) and fluorescence microscopy images of hypothalamic slice preparations. (C) Representative trace of electrophysiological recordings from mCherry positive DREADDs-expressing MCH neurons before, during, and after clozapine-N-oxide (CNO; the DREADDs ligand) application. (D) Rats were bilaterally injected in the lateral hypothalamic area and zona incerta with an AAV driving hM3Gq DREADDS expression under the control of a promoter for MCH (AAV2-rMCHp-hM3D(Gq)-mCherry). Data from electrophysiological recordings showing that CNO significantly increased the firing rate of mCherry positive DREADDs-expressing MCH neurons (n=8). (E) CNO (18 mmoles) was used to pharmacologically activate MCH neurons, which resulted in increased feeding that was blocked by pretreatment with the MCH 1R antagonist H6408 (ANT; 10μg)(n=7,8/group) (*P<.05). All data are represented as means ± SEM. mt, mammillothalamic tract; fx, fornix; V3, third ventricle.