Yang, Su-Rong Hu, Zhen-Zhen Luo, Yan-Jia Zhao, Ya-Nan Sun, Huan-Xin Yin, Dou Wang, Chen-Yao Yan, Yu-Dong Wang, Dian-Ru Yuan, Xiang-Shan Ye, Chen-Bo Guo, Wei Qu, Wei-Min Cherasse, Yoan Lazarus, Michael Ding, Yu-Qiang Huang, Zhi-Li Pharmacogenetic activation of the hM3Dq-expressing rat rostromedial tegmental nucleus (RMTg) neurons. <p>Schematic representation of the Cre-independent adeno-associated virus (AAV) vectors expressing hM3Dq receptors under the control of hSyn promoter. hSyn, human synapsin promoter; WPRE, woodchuck hepatitis virus post-transcriptional regulatory element; CNO, clozapine-N-oxide; IP, intraperitoneal. (B) The coronal section shows the superimposed virus-injected area in 9 rats numbered with the same color characters as the closed curves. The bilateral shaded areas indicate the rat RMTg locations. (C) mCherry immunolabeling (red) indicates hM3Dq-expressing neurons of the RMTg. IPN, interpeduncular nucleus; ml, medial lemniscus. (D) Representative photomicrographs of the RMTg depicting mCherry expression (red), γ-amino butyric acid (GABA) immunoreactivity (green), 4,6-diamidino-2-phenylindole (DAPI, blue), and merge (yellow) images from a rat microinjected with Cre-independent AAV vectors containing hM3Dq. (E, F) CNO (E), not saline (F), drives c-Fos expression in hM3Dq-expressing neurons in the RMTg. (G) Quantification of c-Fos staining in mCherry-expressing RMTg neurons after saline or CNO injection in rats. **<i>p</i> < 0.01 versus saline by unpaired <i>t</i> tests (<i>n</i> = 3, per group). (H) A typical example of voltage trace recorded from an hM3Dq<sup>+</sup> rat RMTg neuron during application of CNO in a brain slice. CNO application (horizontal bar) increased the action potential firing. (I) Merged image of mCherry and biocytin showing a fluorescent view of the patched cell. Left panel: biocytin-labeled neuron; right panel: cell with green (biocytin; top), red (hM3Dq-mCherry; middle), and yellow (merged; bottom) fluorescence. (J, K) Average membrane potential of RMTg hM3Dq<sup>+</sup> neurons (J, <i>n</i> = 7 cells), but not hM3Dq<sup>−</sup> neurons (K, <i>n</i> = 5 cells), was significantly increased by CNO application. The results from each cell are shown on the scatterplot. **<i>p</i> < 0.01 versus saline by paired <i>t</i> tests. Underlying data can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2002909#pbio.2002909.s001" target="_blank">S1 Data</a>.</p> control rats;2- hour recovery period;midbrain dopaminergic system;baseline levels;rat RMTg neurons;midbrain dopaminergic neurons;ventral tegmental area;pharmacogenetic;VTA;NREM;rostromedial tegmental nucleus;SWA;non-rapid eye movement;lesion;dorsal raphe nucleus 2018-04-13
    https://plos.figshare.com/articles/figure/Pharmacogenetic_activation_of_the_hM3Dq-expressing_rat_rostromedial_tegmental_nucleus_RMTg_neurons_/6141428
10.1371/journal.pbio.2002909.g001