Specific and behaviorally consequential astrocyte Gq GPCR signaling attenuation in vivo with iβARK

Jun Nagai; Arash Bellafard; Zhe Qu; Xinzhu Yu; Matthias Ollivier; Mohitkumar R Gangwani; Blanca Diaz-Castro; Giovanni Coppola; Sarah M Schumacher; Peyman Golshani; Viviana Gradinaru; Baljit S Khakh

doi:10.1016/j.neuron.2021.05.023

Specific and behaviorally consequential astrocyte G_q GPCR signaling attenuation in vivo with iβARK

Neuron. 2021 Jul 21;109(14):2256-2274.e9. doi: 10.1016/j.neuron.2021.05.023. Epub 2021 Jun 16.

Authors

Affiliations

¹ Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; RIKEN Center for Brain Science, 2-1 Hirosawa Wako City, Saitama 351-0198, Japan.
² Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA.
³ Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
⁴ Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, 514 Burrill Hall, 407 S. Goodwin Ave, Urbana, IL 61801, USA.
⁵ Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA.
⁶ Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Center for Neurobehavioral Genetics, Semel Institute for Neuroscience and Human Behavior, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA.
⁷ Department of Cardiovascular & Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
⁸ Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Center for Neurobehavioral Genetics, Semel Institute for Neuroscience and Human Behavior, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Integrative Center for Learning and Memory, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; West LA Veterans Affairs Medical Center, Los Angeles, CA 90073, USA; Intellectual and Developmental Disabilities Research Center, Los Angeles, CA, USA.
⁹ Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095-1751, USA; Department of Neurobiology, University of California, David Geffen School of Medicine, Los Angeles, Los Angeles, CA 90095-1751, USA. Electronic address: bkhakh@mednet.ucla.edu.

Abstract

Astrocytes respond to neurotransmitters and neuromodulators using G-protein-coupled receptors (GPCRs) to mediate physiological responses. Despite their importance, there has been no method to genetically, specifically, and effectively attenuate astrocyte G_q GPCR pathways to explore consequences of this prevalent signaling mechanism in vivo. We report a 122-residue inhibitory peptide from β-adrenergic receptor kinase 1 (iβARK; and inactive D110A control) to attenuate astrocyte G_q GPCR signaling. iβARK significantly attenuated G_q GPCR Ca²⁺ signaling in brain slices and, in vivo, altered behavioral responses, spared other GPCR responses, and did not alter astrocyte spontaneous Ca²⁺ signals, morphology, electrophysiological properties, or gene expression in the striatum. Furthermore, brain-wide attenuation of astrocyte G_q GPCR signaling with iβARK using PHP.eB adeno-associated viruses (AAVs), when combined with c-Fos mapping, suggested nuclei-specific contributions to behavioral adaptation and spatial memory. iβARK extends the toolkit needed to explore functions of astrocyte G_q GPCR signaling within neural circuits in vivo.

Keywords: AAV; GPCR; Gq; astrocyte; behavior; behavioral adaptation; calcium; signaling; silencing; spatial memory.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Astrocytes / metabolism*
Brain / metabolism*
Calcium / metabolism
Mice
Neurons / metabolism
Receptors, G-Protein-Coupled / metabolism*
Signal Transduction / physiology*
beta-Adrenergic Receptor Kinases / metabolism*

Substances

Receptors, G-Protein-Coupled
beta-Adrenergic Receptor Kinases
Calcium

Abstract

Publication types

MeSH terms

Substances

Grants and funding