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Cytosolic Ca2+ shifts as early markers of cytotoxicity

Philippe Wyrsch1, Christian Blenn, Theresa Pesch

  • 1Institute of Pharmacology and Toxicology, University of Zurich-Vetsuisse, Winterthurerstrasse 260, Zurich, CH-8057, Switzerland. felix.althaus@vetpharm.uzh.ch.

Cell Communication and Signaling : CCS
|February 7, 2013
PubMed

Insights

Assessing compound toxicity is crucial for risk assessment. This study reveals that monitoring changes in free cytosolic calcium (Ca2+) offers a rapid and sensitive method for predicting cell fate, outperforming traditional viability assays.

Area of Science:

  • Toxicology
  • Cell Biology
  • Biochemistry

Background:

  • Cytotoxicity assessment is vital for risk evaluation of novel compounds.
  • Existing methods rely on metabolic, bio-energetic, or morphological endpoints.
  • Predicting cell fate requires understanding toxin-cell interactions.

Purpose of the Study:

  • To evaluate cytosolic calcium (Ca2+) homeostasis alterations as a cytotoxicity endpoint.
  • To compare Ca2+ flux measurements with the Alamar Blue cell viability assay.
  • To establish a rapid and sensitive cytotoxicity assay for high-throughput screening.

Main Methods:

  • Utilized Fluo-4 dye to measure free cytosolic Ca2+ in mammalian cell lines.
  • Tested a panel of toxins including As2O3, gossypol, H2O2, staurosporine, and titanium(IV)-salane complexes.
  • Correlated Ca2+ shifts with Alamar Blue EC25/EC75 values and caspase/PARP activity.

Main Results:

  • All tested toxins induced rapid (within 5 s) and persistent cytosolic Ca2+ increases.
  • Ca2+ shifts were observed independently of toxin structure and cell death mode.
  • Early Ca2+ flux correlated linearly with Alamar Blue assay results.

Conclusions:

  • Cytosolic Ca2+ monitoring provides a sensitive cytotoxicity assay with a low limit of detection.
  • This Ca2+ flux assay is significantly faster (at least 24x) than Alamar Blue.
  • The assay is suitable for high-throughput screening of compound toxicity.

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