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

Biochem Biophys Res Commun. 2009 Mar 20;380(4):724-8. doi: 10.1016/j.bbrc.2009.01.128. Epub 2009 Jan 27.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Fluorescence Recovery After Photobleaching / methods
  • Fluorescence Resonance Energy Transfer / methods*
  • Green Fluorescent Proteins
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Humans
  • Microscopy, Fluorescence / methods
  • Neurons / metabolism*
  • Serotonin Plasma Membrane Transport Proteins / metabolism*

Substances

  • SLC6A4 protein, human
  • Serotonin Plasma Membrane Transport Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins