Protocol to downregulate GABAergic-astrocyte signaling via astrocyte-selective ablation of GABAB receptor in adult mice

Sara Mederos; Andrea Sánchez-Ruiz; Gertrudis Perea

doi:10.1016/j.xpro.2022.101667

Protocol to downregulate GABAergic-astrocyte signaling via astrocyte-selective ablation of GABA_B receptor in adult mice

STAR Protoc. 2022 Dec 16;3(4):101667. doi: 10.1016/j.xpro.2022.101667. Epub 2022 Sep 13.

Authors

Sara Mederos¹, Andrea Sánchez-Ruiz², Gertrudis Perea³

Affiliations

¹ Sainsbury Wellcome Centre for Neural Circuits and Behaviour, University College London, London W1T 4JG, UK. Electronic address: s.mederos@ucl.ac.uk.
² Neuron-Glia Networks Lab. Cajal Institute, CSIC, Madrid 28002, Spain.
³ Neuron-Glia Networks Lab. Cajal Institute, CSIC, Madrid 28002, Spain. Electronic address: gperea@cajal.csic.es.

Abstract

Here, we present a protocol to selectively downregulate GABA_B receptor (GABA_BR) expression in astrocytes of mouse medial prefrontal cortex (mPFC). We first describe the procedure of surgeries and viral injections. We then detail genetic, histological, and functional characterizations of astrocytic GABA_BR ablation using RT-PCR, imaging, and behavioral assays. The use of GABA_B flox mice can be easily adapted to generate astrocyte-selective GABA_BR ablation in different brain areas and postnatal stages, leading to local downregulation of GABAergic-astrocyte signaling without developmental issues. For complete details on the use and execution of this protocol, please refer to Mederos et al. (2021).

Keywords: Behavior; Gene Expression; Microscopy; Model Organisms; Molecular Biology; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Astrocytes* / metabolism
Mice
Prefrontal Cortex / metabolism
Receptors, GABA-B* / metabolism
Signal Transduction
gamma-Aminobutyric Acid / metabolism

Substances

Receptors, GABA-B
gamma-Aminobutyric Acid