Fig. 7: LC NA neurons innervated by VP Glu neurons project to the lateral hypothalamus.

a Schematic diagram of virus injection. Anterograde trans-synaptic tracer AAV1-EF1α-DIO-Flp was injected into the VP in DBH-Cre mice. Two weeks later, AAV-hSyn-fDIO-eYFP was injected into the LC to label LC NA neurons innervated by VP glutaminergic neurons. b, c Representative images and summary of eYFP-labeled neurons stained with TH-antibody in the LC. n = 3 mice. d Representative images showing that downstream nuclei of LC NA neurons innervated by VP Glu neurons include the dorsal part and the medial posterolateral part of the bed nucleus of stria terminalis (BNST), the lateral hypothalamus (LH), the central amygdala (CeA), and the subthalamic parafascicular nucleus (SPF). e Schematic diagram of virus injection and optical fiber implantation. f Representative images of virus expression in the VP, LC, and LH, and locations of optical fibers in the LC and LH. g−j Heat map, averaged traces, peak, and area under curve (AUC) of peak of fluorescent signal in the LH of NE2M mice in response to optogenetic stimulation of the VP-LC Glu projection by i.p. injection of normal saline or CNO. n = 18 trials from 5 NE2M mice. k−m Representative traces, time course (blue, Guanfacine perfusion, n = 13; black, no drug application, n = 8, from 3 mice), and summary of firing in LH neurons before, during, and after bath perfusion of 100 μM guanfacine, an α2a receptor agonist. n A Pie chart showing the percentage of LH neurons inhibited by guanfacine. Two-tailed paired t tests in (i, j). One-way repeated measures ANOVA in (m). P-values are two-sided. Mean ± SEM in (c, i, j, l). Mean in histogram in (m). The source data are in the Supplementary Dataset, Source Data.xlsx. Statistical results are in Supplementary Table 1.