A Biophysical Approach for the Study of Dopamine Receptor Oligomerization
The ability of certain neurotransmitter receptors to form oligomers provides an additional level of fine-tuning of intracellular signaling. Among the techniques allowing study of receptor oligomerization as well as influence of specific ligands on these processes, a biophysical approach with the use of fluorescently tagged receptors is the most sensitive. Measurement of the fluorescence resonance energy transfer (FRET) phenomenon between two fluorescently tagged receptors is considered a very useful and measurable tool to study the physical interactions between receptors either in a single cell or in a population of living cells. Here we describe the use of FRET measurement specifically to monitor protein oligomer formation between dopamine D1R and D2R, but the same methodology can be used to study other receptor proteins as well as their mutants.
Key wordsFRET Neurotransmitters Oligomer formation D1R D2R ECFP EYFP
The authors would like to dedicate this work to the memory of the late professor Zygmunt Wasylewski, who encouraged us to employ fluorescence spectroscopy in our studies of dopamine receptors.