Detecting G Protein-coupled Receptor Complexes in Postmortem Human Brain with Proximity Ligation Assay and a Bayesian Classifier

Overview

Journal Biotechniques

Specialties Biomedical Engineering
Biotechnology

Date 2019 Dec 21

PMID 31859535

Citations 8

Authors

Ying Zhu

Jozsef Meszaros

Roman Walle

Rongxi Fan

Ziyi Sun

Andrew J Dwork

Pierre Trifilieff

Jonathan A Javitch

Affiliations

Soon will be listed here.

Abstract

Despite the controversy regarding the existence and physiological relevance of class A G protein-coupled receptor dimerization, there is substantial evidence for functional interactions between the dopamine D2 receptor (D2R) and the adenosine A2A receptor (A2AR). A2AR-D2R complexes have been detected in rodent brains by proximity ligation assay; however, their existence in the human brain has not been demonstrated. In this study, we used Brightfield proximity ligation assay, combined with a systematic sampling and a parameter-free naive Bayesian classifier, and demonstrated proximity between the D2R and the A2AR in the adult human ventral striatum, consistent with their colocalization within complexes and the possible existence of D2R-A2AR heteromers. These methods are applicable to the relative quantification of proximity of two proteins, as well as the expression levels of individual proteins.

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