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Visualizing physiological parameters in cells and tissues using genetically encoded indicators for metabolites.

A San Martín1, R Arce-Molina1, C Aburto2

  • 1Centro de Estudios Científicos (CECs), Valdivia, Chile.

Free Radical Biology & Medicine
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Metabolic research is advancing with genetically-encoded fluorescent indicators (GEFIs) that visualize metabolite dynamics in living cells. These tools offer high resolution for studying metabolic pathways and cellular functions.

Keywords:
FRETFluorescenceFluxesMetabolitesSingle-fluorophore

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Area of Science:

  • Cellular Metabolism
  • Biochemistry
  • Microscopy

Background:

  • Metabolism research is rapidly evolving with new technologies.
  • Over 50 genetically-encoded fluorescent indicators (GEFIs) have been developed since 2002.
  • GEFIs enable high spatial and temporal resolution monitoring of metabolites using fluorescence microscopy.

Purpose of the Study:

  • To review genetically-encoded fluorescent indicators (GEFIs) validated in mammalian systems.
  • To highlight the capabilities of GEFIs for monitoring various metabolites.
  • To emphasize the importance of proper data handling for GEFI applications.

Main Methods:

  • Survey of validated GEFIs in mammalian systems.
  • Description of GEFI mechanism (fusion proteins changing fluorescence upon metabolite binding).
  • Discussion of GEFI applications for monitoring sugars, monocarboxylates, Krebs cycle intermediates, amino acids, cofactors, and energy nucleotides.

Main Results:

  • GEFIs allow monitoring of relative metabolite levels, concentrations, and fluxes in living systems.
  • Some GEFIs can provide absolute concentration data with calibration.
  • These indicators are minimally invasive and crucial for metabolic research.

Conclusions:

  • GEFIs are powerful tools for metabolic research, enabling confirmation, rebuttal, and discovery.
  • Accurate data collection, processing, and interpretation are essential for effective use of GEFIs.
  • This technology is expected to significantly advance metabolic physiology.