Flow cytometry application for studies on adenosine A2A receptors expression

Adenosine A(2A) receptors belong to the heptaspanning membrane receptors family A, also known as G protein-coupled receptors. In human brain they are highly expressed in striatum, where they co-exist and co-function with adenosine A(1), glutamate mGlu(5) and dopamine D-2 receptors. As glutaminergic neurotransmission modulators in GABAergic enkephalinergic neurons, adenosine A(2A) receptors are attractive targets for new, alternative therapies of neurodegenerative disorders, like Parkinson's disease and Huntington's disease. The aim of the research was to obtained fluorescently tagged adenosine A(2A) receptors. Gene encoding human adenosine A(2A) receptor was inserted into plasmid pEYFP-N1, bearing enhanced yellow fluorescent protein (EYFP). The construct was expressed in HEK 293 cells. Fluorescence was observed by flow cytometry and epifluorescence microscopy. Functional ligand binding properties were investigated by saturation binding analysis of adenosine A(2A) receptors specific agonist [H-3] CGS 21680.