C 4, densities of DR Tph2→SC and →AC neurons along D–V axis. C 3, coronal projection showing the location of individual cells from the DR Tph2→SC and →AC groups. (C) Merged surface view of the DR Tph2→SC (subcortical) cluster (cyan), →AC (anterior cortex) cluster (red), and →ENT cluster (brown) in coronal (C 1) and sagittal (C 2) view. Right insets (top, coronal view bottom, sagittal view): yellow, cyan, and red structures represent 3D surface of the clusters of, respectively, all DR Tph2 + neurons those that project to PVH/CeA/LHb/dLGN or those that project to OB/PIR/OFC/ENT. Left insets, high-magnification images of neurons marked by arrows in individual channels. Magenta, retrogradely labeled cells from eight projecting sites yellow, anti-Tph2 staining. (B) Representative coronal confocal sections of the DR. DBSCAN was applied for spatial clustering to generate a 3D surface based on the location of Tph2 + neurons (STAR Methods). Each section was registered to the Allen reference brain and reconstructed in 3D. The positions of Tph2 +/tdTomato + cells were recorded in confocal images. Anti-Tph2 staining was performed on consecutive coronal sections containing DR (star). Cyan and red dots represent injection sites of HSV-Cre in Ai14 mice. (A) Schematic of retrograde labeling and 3D reconstruction of spatial locations of DR serotonin neurons. ![]() These results provide compelling evidence that the DR serotonin system contains parallel sub-systems that differ in input and output connectivity, physiological response properties, and behavioral functions.ĥ-HT Tph2 Vglut3 anxiety central amygdala depression dorsal raphe fiber photometry iDISCO orbital frontal cortex serotonin.Ĭopyright © 2018 Elsevier Inc. ![]() Gain- and loss-of-function experiments suggest that amygdala-projecting DR serotonin neurons promote anxiety-like behavior, whereas frontal-cortex-projecting neurons promote active coping in the face of challenge. Further, amygdala- and frontal-cortex-projecting DR serotonin neurons have largely complementary whole-brain collateralization patterns, receive biased inputs from presynaptic partners, and exhibit opposite responses to aversive stimuli. Using viral-genetic methods, we found that subcortical- and cortical-projecting serotonin neurons have distinct cell-body distributions within the DR and differentially co-express a vesicular glutamate transporter. Most functional studies to date have treated DR serotonin neurons as a single population. The dorsal raphe (DR) constitutes a major serotonergic input to the forebrain and modulates diverse functions and brain states, including mood, anxiety, and sensory and motor functions.
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