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ACh signaling modulates activity of the GABAergic signaling network in the basolateral amygdala and behavior in stress-relevant paradigms

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Abstract

The balance between excitatory and inhibitory (E/I) signaling is important for maintaining homeostatic function in the brain. Indeed, dysregulation of inhibitory GABA interneurons in the amygdala has been implicated in human mood disorders. We hypothesized that acetylcholine (ACh) signaling in the basolateral amygdala (BLA) might alter E/I balance resulting in changes in stress-sensitive behaviors. We therefore measured ACh release as well as activity of calmodulin-dependent protein kinase II (CAMKII)-, parvalbumin (PV)-, somatostatin (SOM)- and vasoactive intestinal protein (VIP)-expressing neurons in the BLA of awake, behaving male mice. ACh levels and activity of both excitatory and inhibitory BLA neurons increased when animals were actively coping, and decreased during passive coping, in the light–dark box, tail suspension and social defeat. Changes in neuronal activity preceded behavioral state transitions, suggesting that BLA activity may drive the shift in coping strategy. In contrast to exposure to escapable stressors, prolonging ACh signaling with a cholinesterase antagonist changed the balance of activity among BLA cell types, significantly increasing activity of VIP neurons and decreasing activity of SOM cells, with little effect on CaMKII or PV neurons. Knockdown of α7 or β2-containing nAChR subtypes in PV and SOM, but not CaMKII or VIP, BLA neurons altered behavioral responses to stressors, suggesting that ACh signaling through nAChRs on GABA neuron subtypes contributes to stress-induced changes in behavior. These studies show that ACh modulates the GABAergic signaling network in the BLA, shifting the balance between SOM, PV, VIP and CaMKII neurons, which are normally activated coordinately during active coping in response to stress. Thus, prolonging ACh signaling, as occurs in response to chronic stress, may contribute to maladaptive behaviors by shifting the balance of inhibitory signaling in the BLA.

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Fig. 1: Activity of BLA neurons and ACh release during stress-relevant behaviors.
Fig. 2: GCaMP6f activity in CaMKII, VIP, PV and SOM neurons following physostigmine administration (0.15 mg/kg, i.p.).
Fig. 3: Knockdown of the α7 or β2 nAChR subunits in principal neurons (CAMKII-Cre), inhibitory neurons (GAD65-Cre) or GABAergic neuronal subtypes (VIP-Cre, PV-Cre, SOM-Cre) in the BLA.
Fig. 4: Schematic summarizing effects of ACh and cholinergic receptors in the BLA and their influence on behavior.

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Acknowledgements

These studies were supported by National Institutes of Health grants MH077681, MH105824, and DA033945 from the National Institutes of Health and a NARSAD Distinguished Investigator grant from the Brain and Behavior Research Foundation. This work was funded in part by the State of Connecticut, Department of Mental Health and Addiction Services, but this publication does not express the views of the Department of Mental Health and Addiction Services or the State of Connecticut.

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  1. Seth R. Taylor

    Present address: Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

Authors and Affiliations

  1. Department of Psychiatry, Yale University School of Medicine, 34 Park Street, 3rd Floor Research, New Haven, CT, 06508, USA

    Yann S. Mineur, Tenna N. Mose, Kathrine Lefoli Maibom, Steven T. Pittenger, Alexa R. Soares, Hao Wu, Seth R. Taylor, Yaqing Huang & Marina R. Picciotto

  2. Interdepartmental Neuroscience Program, Yale University School of Medicine, 34 Park Street, 3rd Floor Research, New Haven, CT, 06508, USA

    Alexa R. Soares, Hao Wu, Seth R. Taylor & Marina R. Picciotto

  3. Experimental Pathology Graduate Program, Yale University School of Medicine, 34 Park Street, 3rd Floor Research, New Haven, CT, 06508, USA

    Yaqing Huang

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Contributions

YSM designed, carried out and contributed to all experiments, analyzed data, and wrote the manuscript. TNM and STP carried out and contributed to all knockdown experiments; KLM carried out and contributed to knockdown experiments and GCaMP fiber photometry recording and analyses; ARS contributed to fiber photometry analyses; HW and YH carried out and contributed to GACh fiber photometry experiments and analyses; SRT designed and validated the conditional nAChR knockdown vector; MRP designed the study, analyzed outcomes, and wrote the manuscript. All authors reviewed and approved the manuscript.

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Correspondence to Marina R. Picciotto.

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Mineur, Y.S., Mose, T.N., Maibom, K.L. et al. ACh signaling modulates activity of the GABAergic signaling network in the basolateral amygdala and behavior in stress-relevant paradigms. Mol Psychiatry 27, 4918–4927 (2022). https://doi.org/10.1038/s41380-022-01749-7

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