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PAC1 Receptor Internalization and Endosomal MEK/ERK Activation Is Essential for PACAP-Mediated Neuronal Excitability

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Abstract

Pituitary adenylate cyclase activating polypeptide (PACAP, Adcyap1) activation of PAC1 receptors (Adcyap1r1) can significantly increase the excitability of diverse neurons through differential mechanisms. For guinea pig cardiac neurons, the modulation of excitability can be mediated in part by PAC1 receptor plasma membrane G protein-dependent activation of adenylyl cyclase and downstream signaling cascades. By contrast, PAC1 receptor-mediated excitability of hippocampal dentate gyrus granule cells appears independent of membrane-delimited AC/cAMP/PKA and PLC/PKC signaling. For both neuronal types, there is mechanistic convergence demonstrating that endosomal PAC1 receptor signaling has prominent roles. In these models, neuronal exposure to Pitstop2 to inhibit β-arrestin/clathrin-mediated PAC1 receptor internalization eliminates PACAP modulation of excitability. β-arrestin is a scaffold for a number of effectors especially MEK/ERK and notably, paradigms that inhibit PAC1 receptor endosome formation and ERK signaling also blunt the PACAP-induced increase in excitability. Detailed PAC1 receptor internalization and endosomal ERK signaling mechanisms have been confirmed in HEK PAC1R-EGFP cells and shown to be long lasting which appear to recapitulate the sustained electrophysiological responses. Thus, PAC1 receptor internalization/endosomal recruitment efficiently and efficaciously activates MEK/ERK signaling and appears to represent a singular and critical common denominator in regulating neuronal excitability by PACAP.

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Acknowledgements

We thank Thomas Buttolph for excellent technical assistance.

Funding

This work was supported in part by National Institutes of Health (NIH) grant National Institute of General Medical Sciences (NIGMS) P30 GM103498/National Center for Research Resources (NCRR) P30 RR032135 (RLP) and National Institute of Mental Health (NIMH) MH097988 (SEH and VM).

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Authors and Affiliations

  1. Departmental of Neurological Sciences, University of Vermont College of Medicine, Burlington, VT, USA

    Victor May, Karen M. Braas & Rodney L. Parsons

  2. Department of Psychological Science, University of Vermont, Burlington, VT, USA

    Gregory C. Johnson & Sayamwong E. Hammack

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  1. Victor May
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  2. Gregory C. Johnson
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  3. Sayamwong E. Hammack
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RLP and VM wrote the paper; GCJ, SEH, and KMB edited and revised the manuscript. VM and KMB generated data and created figures.

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Correspondence to Victor May.

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May, V., Johnson, G.C., Hammack, S.E. et al. PAC1 Receptor Internalization and Endosomal MEK/ERK Activation Is Essential for PACAP-Mediated Neuronal Excitability. J Mol Neurosci 71, 1536–1542 (2021). https://doi.org/10.1007/s12031-021-01821-x

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