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Subcellular Imaging of Neuronal Calcium Handling In Vivo
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Measuring baseline Ca(2+) levels in subcellular compartments using genetically engineered fluorescent indicators.

Julia M Hill1, Diego De Stefani2, Aleck W E Jones1

  • 1Department of Cell and Developmental Biology, Consortium for Mitochondrial Research, University College London, London, United Kingdom.

Methods in Enzymology
|June 14, 2014
PubMed
Summary
This summary is machine-generated.

This study reviews genetically engineered fluorescent calcium (Ca2+) probes for measuring organelle-specific Ca2+ levels. These tools help assess how basal Ca2+ impacts cell metabolism, particularly in cancer cells.

Keywords:
Endoplasmic reticulumFRETGenetically encoded Ca(2+) indicatorMitochondriaOrganelle targetingRecombinant fluorescent Ca(2+) probe

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Intracellular calcium (Ca2+) signaling is crucial for physiological and pathological processes.
  • A crosstalk exists between cellular bioenergetic metabolism and Ca2+ homeostasis, influencing cell fate.
  • Endoplasmic reticulum and mitochondria are key organelles for metabolism and Ca2+ signaling, but localized fluxes are hard to detect.

Purpose of the Study:

  • To provide an overview of genetically engineered, organelle-targeted fluorescent Ca2+ probes and their evolution.
  • To describe protocols for quantifying baseline Ca2+ concentrations in specific subcellular compartments.
  • To highlight the application of these methods in assessing the metabolic profile of cancer cells based on basal Ca2+ levels.

Main Methods:

  • Development and application of organelle-targeted fluorescent Ca2+ indicators.
  • Utilizing genetically engineered probes for specific subcellular compartment targeting.
  • Establishing protocols to quantify baseline Ca2+ concentrations in organelles.

Main Results:

  • Organelle-targeted fluorescent Ca2+ probes have evolved significantly over two decades.
  • Protocols for quantifying basal Ca2+ in specific subcellular compartments have been developed.
  • These methods enable assessment of the impact of basal Ca2+ on cellular metabolism.

Conclusions:

  • Genetically engineered, organelle-targeted fluorescent Ca2+ probes are valuable tools for studying cellular physiology.
  • Quantifying basal Ca2+ concentrations in organelles is vital for understanding metabolic functions.
  • This approach is suitable for investigating metabolic alterations in cancer cells.