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Author Spotlight: Fluorescence-Based Quantification of Mitochondrial Membrane Potential and Superoxide Levels Using Live Imaging in HeLa Cells
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Mitochondrial Ca2+ concentrations in live cells: quantification methods and discrepancies.

Celia Fernandez-Sanz1, Sergio De la Fuente1, Shey-Shing Sheu1

  • 1Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA, USA.

FEBS Letters
|May 7, 2019
PubMed
Summary

Accurately measuring mitochondrial calcium (Ca2+) is crucial for understanding cell function. This review highlights discrepancies in current methods for quantifying mitochondrial Ca2+ levels and suggests future research directions.

Keywords:
fluorescent Ca2+ indicatorsgenetically encoded Ca2+ indicatorslive cellsmitochondrial Ca2+ concentrations

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Area of Science:

  • Cellular biology
  • Mitochondrial function
  • Calcium signaling

Background:

  • Intracellular calcium (Ca2+) regulates vital cellular processes.
  • Mitochondria are key players in Ca2+ signaling, influencing cellular functions and Ca2+ dynamics.
  • Accurate measurement of mitochondrial Ca2+ ([Ca2+]m) is essential for understanding these processes.

Purpose of the Study:

  • To review and summarize reported calibrated mitochondrial Ca2+ ([Ca2+]m) values across various cell types.
  • To discuss the significant discrepancies observed in [Ca2+]m measurements using different methodologies.
  • To identify areas for future research to improve the precision of [Ca2+]m determination.

Main Methods:

  • Literature review of studies reporting calibrated mitochondrial Ca2+ ([Ca2+]m) measurements.
  • Analysis of discrepancies arising from distinct measurement techniques, Ca2+ ratios, and cell-specific dynamics.
  • Synthesis of existing data to provide a comprehensive overview of [Ca2+]m values.

Main Results:

  • Significant discrepancies exist in the reported absolute [Ca2+]m values in stimulated live cells.
  • Methodological differences, Ca2+ binding properties, and cell/stimulus-dependent dynamics contribute to these variations.
  • Existing data reveal a wide range of [Ca2+]m values depending on the experimental approach.

Conclusions:

  • Precise determination of absolute [Ca2+]m is critical for understanding Ca2+ signaling and mitochondrial function.
  • Addressing the discrepancies in measurement techniques is imperative for advancing the field.
  • Further research is needed to establish standardized and reliable methods for quantifying [Ca2+]m.