Live-cell imaging of rapid calcium dynamics using fluorescent, genetically-encoded GCaMP probes with Aspergillus fumigatus

Fungal Genet Biol. 2021 Jun:151:103470. doi: 10.1016/j.fgb.2020.103470. Epub 2020 Sep 24.

Abstract

Calcium signalling plays a fundamental role in fungal intracellular signalling. Previous approaches (fluorescent dyes, bioluminescent aequorin, genetically encoded cameleon probes) with imaging rapid subcellular changes in cytosolic free calcium ([Ca2+]c) in fungal cells have produced inconsistent results. Recent data obtained with new fluorescent, genetically encoded GCaMP probes, that are very bright, have resolved this problem. Here, exposing conidia or conidial germlings to high external Ca2+, as an example of an external stressor, induced very dramatic, rapid and dynamic [Ca2+]c changes with localized [Ca2+]c transients and waves. Considerable heterogeneity in the timing of Ca2+ responses of different spores/germlings within the cell population was observed.

Keywords: Aspergillus fumigatus; Calcium imaging; GCaMP calcium probes; Live-cell imaging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus fumigatus / metabolism*
  • Calcium / metabolism*
  • Calcium Signaling
  • Calmodulin / genetics
  • Calmodulin / metabolism
  • Fluorescent Dyes / metabolism*
  • Genes, Reporter*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Molecular Probes
  • Myosin-Light-Chain Kinase / genetics
  • Myosin-Light-Chain Kinase / metabolism
  • Spores, Fungal / metabolism

Substances

  • Calmodulin
  • Fluorescent Dyes
  • Molecular Probes
  • Green Fluorescent Proteins
  • Myosin-Light-Chain Kinase
  • Calcium