In situ visualization of opioid and cannabinoid drug effects using phosphosite-specific GPCR antibodies.

ABSTRACT: G protein-coupled receptors (GPCRs) are important signal transducers that are phosphorylated upon activation at intracellular serine and threonine residues. Although antibodies that specifically recognize the phosphorylation state of GPCRs have been available for many years, efficient immunolocalization of phosphorylated receptors in their tissues of origin has not been possible. Here, we show that phosphorylation of receptors is highly unstable during routine immunohistochemical procedures, requiring the use of appropriate phosphatase inhibitors particular during tissue perfusion, post-fixation, and cryoprotection but not during immunostaining of tissue sections. We provide proof of concept using phosphorylation state-specific μ-opioid receptor (MOP) and cannabinoid receptor 1 (CB1) antib

SUBMITTER: Fritzwanker S 

PROVIDER: S-EPMC10105690 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature