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Activation of Lateral Parabrachial Nucleus (LPBn) PACAP-Expressing Projection Neurons to the Bed Nucleus of the Stria Terminalis (BNST) Enhances Anxiety-like Behavior

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Abstract

Anxiety disorders are among the most common psychiatric disorders, and understanding the underlying neurocircuitry of anxiety- and stress-related behaviors may be important for treatment. The bed nucleus of the stria terminalis (BNST) has been studied for its role in many stress-related pathologies, such as anxiety, pain, depression, and addiction. Our prior work has demonstrated that pituitary adenylate cyclase-activating polypeptide (PACAP) receptor activation in the BNST mediates many of the behavioral consequences of chronic stress. While the BNST contains local PACAP-expressing neurons, a major source of afferent PACAP is the lateral parabrachial nucleus (LPBn), and excitotoxic lesions of the LPBn substantially decreasess PACAP immunostaining in the BNST. Here, we first assessed Cre-dependent reporter expression by injecting AAV2-hSyn-DIO-mCherry into the LPBn of PACAP-IRES-Cre mice for circuit mapping studies and identified PACAP projections to the BNST, lateral capsular central nucleus of the amygdala (CeLC), and ventromedial hypothalamus (VMH). In a second study, we assessed the effects of chemogenetically activating LPBn PACAP afferents in the BNST by injecting AAV2-hSyn-DIO-hM3D(Gq)-mCherry into the LPBn of PACAP-IRES-Cre mice for Cre-dependent expression of excitatory designer receptors exclusively activated by designer drugs (DREADDs). Before behavioral testing, clozapine-N-oxide (CNO), the selective agonist of our DREADD, was infused directly into the BNST. We found that after specific activation of LPBn PACAP afferents in the BNST, mice had increased anxiety-like behavior compared with controls, while total locomotor activity was unaffected. These results indicate that activation of PACAPergic LPBn projections to the BNST may play an important role in producing anxiety-like behavior.

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Availability of Data and Materials

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Galen Missig and Matt McCabe for the preliminary viral injection studies.

Funding

This work was supported in part by the National Institutes of Health (NIH) grant and the National Institute of Mental Health (NIMH) MH097988 (SEH and VM).

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Authors and Affiliations

  1. Department of Psychological Science, University of Vermont, 2 Colchester Avenue, Burlington, VT, 05405, USA

    Melissa N. Boucher, Mahafuza Aktar, Karen M. Braas, Victor May & Sayamwong E. Hammack

  2. Department of Neurological Sciences, College of Medicine, University of Vermont, 149 Beaumont Avenue, Burlington, VT, 05405, USA

    Melissa N. Boucher, Mahafuza Aktar, Karen M. Braas, Victor May & Sayamwong E. Hammack

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  1. Melissa N. Boucher
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  2. Mahafuza Aktar
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  3. Karen M. Braas
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  4. Victor May
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  5. Sayamwong E. Hammack
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Contributions

MNB and SEH designed and executed experiments. MNB, VM, and SEH contributed to the data analyses; MNB, MA, KMB, VM, and SEH all contributed to the writing and creation of the figures for the manuscript.

Corresponding author

Correspondence to Sayamwong E. Hammack.

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Ethics Approval and Consent to Participate

All the animal procedures were approved by the University of Vermont Institutional Animal Care and Use Committee and carried out with accordance with the National Institutes of Health’s Guide for the Care and Use of Laboratory Animals. No human studies were performed in this article.

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Not applicable.

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The authors declare no competing interests.

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Boucher, M.N., Aktar, M., Braas, K.M. et al. Activation of Lateral Parabrachial Nucleus (LPBn) PACAP-Expressing Projection Neurons to the Bed Nucleus of the Stria Terminalis (BNST) Enhances Anxiety-like Behavior. J Mol Neurosci 72, 451–458 (2022). https://doi.org/10.1007/s12031-021-01946-z

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