Serotonin transporter oligomerization documented in RN46A cells and neurons by sensitized acceptor emission FRET and fluorescence lifetime imaging microscopy

Abstract

Serotonin transporter oligomerization documented in RN46A cells and neurons by sensitized acceptor emission FRET and fluorescence lifetime imaging microscopy

Anja W. Fjorback^1&2, Patrick Pla^3&4, Heidi K. Müller², Ove Wiborg², Frédéric Saudou^3&4, Jens R. Nyengaard¹, and Merete K. Raarup^1*

 

The serotonin transporter is a member of the monoamine transporter family that also includes transporters of dopamine and norepinephrine. We have used sensitized acceptor emission fluorescence resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) to study the oligomerization of SERT in HEK-MSR-239 cells, RN46A cells and in cultured hippocampal neurons. We were able to show identical FRET efficiencies in cell lines as well as in primary cultured hippocampal neurons, demonstrating that the oligomerization is cell type independent. The results obtained with both FRET approaches are very similar and furthermore, in agreement with previous results obtained by donor bleaching FRET microscopy.

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¹Stereology and Electron Microscopy Research Laboratory, HistoInformatics and MIND Center, University of Aarhus, Ole Worms Allé, building 8, DK-8000 Aarhus C, Denmark ²Centre  for Psychiatry Research, University Hospital Aarhus, Skovagervej 2, DK-8240 Risskov, Denmark ^ 3Institute Curie, ⁴CNRS UMR146, Orsay F-91405, France