Assessing CB Expression in the Brain by Immunohistochemical Methods: Light, Confocal, and Electron Microscopy

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2022 Sep 24

PMID 36152206

Authors

Roberta Imperatore

Luigia Cristino

Affiliations

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Abstract

Conventional techniques to reveal the neuroanatomical distribution of type 1 cannabinoid receptor (CB) in the brain, at the cellular and subcellular level, are mainly represented by light, confocal, and electron microscopy. By using immunoperoxidase and immunofluorescence methods, it is possible to reveal CB distribution and localization in the brain and its changes under pathological conditions. Moreover, by using electron microscopy, we can define the ultrastructural localization at the level of subcellular structures and organelles. Here, we describe immunoperoxidase, immunofluorescence, and electron microscopy protocols used to get information about CB spatial distribution and localization in the brain. Preparation of reagents, resin embedding, preparation for an endogenous activity-blocking step, and background counterstaining and revelation of CB by using specific labeled secondary antibodies will be presented. The methods here discussed are highly sensitive and specific multistep processes, where each step is critical to finally obtain an optimum signal.

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