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Ventral tegmental area glutamate neurons mediate nonassociative consequences of stress

Abstract

Exposure to trauma is a risk factor for the development of a number of mood disorders, and may enhance vulnerability to future adverse life events. Recent data demonstrate that ventral tegmental area (VTA) neurons expressing the vesicular glutamate transporter 2 (VGluT2) signal and causally contribute to behaviors that involve aversive or threatening stimuli. However, it is unknown whether VTA VGluT2 neurons regulate transsituational outcomes of stress and whether these neurons are sensitive to stressor controllability. This work adapted an operant mouse paradigm to examine the impact of stressor controllability on VTA VGluT2 neuron function as well as the role of VTA VGluT2 neurons in mediating transsituational stressor outcomes. Uncontrollable (inescapable) stress, but not physically identical controllable (escapable) stress, produced social avoidance and exaggerated fear in male mice. Uncontrollable stress in females led to exploratory avoidance of a novel brightly lit environment. Both controllable and uncontrollable stressors increased VTA VGluT2 neuronal activity, and chemogenetic silencing of VTA VGluT2 neurons prevented the behavioral sequelae of uncontrollable stress in male and female mice. Further, we show that stress activates multiple genetically-distinct subtypes of VTA VGluT2 neurons, especially those that are VGluT2+VGaT+, as well as lateral habenula neurons receiving synaptic input from VTA VGluT2 neurons. Our results provide causal evidence that mice can be used for identifying stressor controllability circuitry and that VTA VGluT2 neurons contribute to transsituational stressor outcomes, such as social avoidance, exaggerated fear, or anxiety-like behavior that are observed within trauma-related disorders.

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Fig. 1: Inescapable stress reduces social exploration in male, but not female, C57BL/6 J mice.
Fig. 2: Stress-induced reduction in social exploration is dependent on the controllability of the stressor in male mice.
Fig. 3: VTA VGluT2 neuron signaling is enhanced by both inescapable and escapable stress.
Fig. 4: hM4Di-receptor activation reduces VTA VGluT2 neuronal activity.
Fig. 5: Chemogenetic suppression of VTA VGluT2 neuron activity blocks nonassociative behavioral consequences of inescapable stress.
Fig. 6: VTA VGluT2 cell-types and pathways activated by inescapable stress.

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Acknowledgements

This research was supported by the Webb-Waring Biomedical Research Award from the Boettcher Foundation (DHR), R01 DA047443 (DHR), F31 MH125569 (DJM), a 2020 NARSAD Young Investigator grant from the Brain and Behavior Research Foundation (DHR), R01 MH050479 (MVB), R21 MH116353 (MVB), American Australian Association Fellowship (MVB), DK078749 (STH), and The University of Colorado Boulder. Ai193 mice were generously provided by the Allen Institute and generated with funds from 1U19MH114830-01. The imaging work was performed at the BioFrontiers Institute Advanced Light Microscopy Core (RRID: SCR_018302). Laser scanning confocal microscopy was performed on a Nikon A1R microscope supported by NIST-CU Cooperative Agreement award number 70NANB15H226. The PerkinElmer Opera Phenix is supported by NIH grant 1S10OD025072. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Prism and Biorender were used to make figures and schematics.

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DJM, MVB, and DHR conceived this project. DJM, AL, KLE, DTH, EDP, SCG, CJM, MVB, and DHR designed, performed, or analyzed behavioral-related experiments. ARR and STH designed, performed, and analyzed whole-cell recording experiments. TLD and BT contributed critical reagents. DJM, MVB, and DHR wrote the manuscript with the contribution of all authors.

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Correspondence to Michael V. Baratta or David H. Root.

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McGovern, D.J., Ly, A., Ecton, K.L. et al. Ventral tegmental area glutamate neurons mediate nonassociative consequences of stress. Mol Psychiatry (2022). https://doi.org/10.1038/s41380-022-01858-3

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