Recording intracellular cAMP levels with EPAC-based FRET sensors by fluorescence lifetime imaging

Methods Mol Biol. 2015:1294:13-24. doi: 10.1007/978-1-4939-2537-7_2.

Abstract

Eukaryotic cells use second messengers such as Ca(2+), IP3, cGMP, and cAMP to transduce extracellular signals like hormones, via membrane receptors to downstream cellular effectors. FRET-based sensors are ideal to visualize and measure these rapid changes of second messenger concentrations in time and place. Here, we describe the use of EPAC-based FRET sensors to measure cAMP with spatiotemporal resolution in cells by fluorescence lifetime imaging (FLIM).

MeSH terms

  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Cell Line, Tumor
  • Cyclic AMP / metabolism*
  • Fluorescence Resonance Energy Transfer / methods
  • Genes, Reporter
  • Humans
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods

Substances

  • Cyclic AMP