Imaging approaches to measuring lysosomal calcium

Methods Cell Biol. 2015:126:159-95. doi: 10.1016/bs.mcb.2014.10.031. Epub 2015 Jan 19.

Abstract

Endolysosomes are emerging as key players that generate as well as respond to intracellular Ca(2+) signals. The role of Ca(2+) in modulating acidic organelle function has long been recognized, but it is now emerging that acidic organelles also act as intracellular Ca(2+) stores; they actively sequester Ca(2+) in their lumina and release it to the cytosol upon activation of endolysosomal Ca(2+) channels. This local Ca(2+) signal is crucial for endolysosomal function and/or global Ca(2+) signaling. Importantly, defects in endolysosomal Ca(2+) are associated with disease. This chapter discusses several complimentary approaches to monitor endolysosomal Ca(2+), with particular emphasis on the inherent pitfalls that can plague the unwary.

Keywords: Bafilomycin A1; Ca(2+); GPN; Lysosome; NAADP; Nigericin; pH.

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Calcium Signaling
  • Cells, Cultured
  • Endosomes / metabolism
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Homeostasis
  • Humans
  • Hydrogen-Ion Concentration
  • Lysosomes / metabolism*
  • Microscopy, Fluorescence
  • Staining and Labeling

Substances

  • Fluorescent Dyes
  • Calcium