Monitoring Calcium Fluxes and Lysosome Exocytosis During Pyroptosis

Methods Mol Biol. 2023:2641:171-178. doi: 10.1007/978-1-0716-3040-2_14.

Abstract

Inflammasome-mediated activation of inflammatory caspases (caspase-1, caspase-4, caspase-5, caspase-11) initiates a cascade of cellular events that lead to proinflammatory cell death, or pyroptosis. Proteolytic cleavage of gasdermin D results in the formation of transmembrane pores that allow the release of mature cytokines IL-1β and IL-18. Gasdermin pores also allow calcium influx through the plasma membrane, triggering the fusion of lysosomal compartments with the cell surface and release of their contents into the extracellular milieu in a process termed lysosome exocytosis. This chapter outlines methods for measuring calcium flux, lysosome exocytosis, and membrane disruption after inflammatory caspase activation.

Keywords: Calcium; Gasdermin D; Inflammation; Inflammatory caspases; Lysosome exocytosis; Pyroptosis.

MeSH terms

  • Calcium* / metabolism
  • Caspase 1 / metabolism
  • Caspases / metabolism
  • Exocytosis
  • Gasdermins
  • Inflammasomes / metabolism
  • Lysosomes / metabolism
  • Pyroptosis*

Substances

  • Calcium
  • Gasdermins
  • Caspases
  • Caspase 1
  • Inflammasomes