Open AccessDetecting G protein-coupled receptor complexes in postmortem human brain with proximity ligation assay and a Bayesian classifier
Author(s) -
Ying Zhu,
József Mészáros,
Roman Walle,
Rongxi Fan,
Ziyin Sun,
Andrew J. Dwork,
Pierre Trifilieff,
Jonathan Javitch
Publication year - 2020
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2019-0083
Subject(s) - proximity ligation assay , colocalization , dopamine receptor d2 , biology , receptor , adenosine a2a receptor , striatum , human brain , g protein coupled receptor , neuroscience , ligation , microbiology and biotechnology , dopamine , biochemistry , adenosine receptor , agonist
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.