G protein-coupled receptor heterodimerization in the brain.
G protein-coupled receptors (GPCRs) play critical roles in cellular processes and signaling and have been shown to form heteromers with diverge biochemical and/or pharmacological activities that are different from those of the corresponding monomers or homomers. However, despite extensive experimental results supporting the formation of GPCR heteromers in heterologous systems, the existence of such receptor heterocomplexes in the brain remains largely unknown, mostly because of the lack of appropriate methodology. Herein, we describe the in situ proximity ligation assay procedure underlining its high selectivity and sensitivity to image GPCR heteromers with confocal microscopy in brain sections. We describe here how the assay is performed and discuss advantages and disadvantages of this method compared with other available techniques.
Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.
Dasiel O
DO
Borroto-Escuela
Wilber
W
Romero-Fernandez
Pere
P
Garriga
Francisco
F
Ciruela
Manuel
M
Narvaez
Alexander O
AO
Tarakanov
Miklós
M
Palkovits
Luigi F
LF
Agnati
Kjell
K
Fuxe
D000494:Allosteric Regulation; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D008958:Models, Molecular; D009473:Neuronal Plasticity; D025941:Protein Interaction Mapping / Q000379:methods; D055503:Protein Multimerization; D017468:Receptors, Fibroblast Growth Factor / Q000032:analysis / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000378:metabolism*; D011985:Receptors, Serotonin / Q000032:analysis / Q000378:metabolism; D015398:Signal Transduction
D017468:Receptors, Fibroblast Growth Factor; D043562:Receptors, G-Protein-Coupled; D011985:Receptors, Serotonin