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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Kayagaki N, Stowe IB, Lee BL et al (2015) Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526:666–671. https://doi.org/10.1038/nature15541
Shi J, Zhao Y, Wang K et al (2015) Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526:660–665. https://doi.org/10.1038/nature15514
Bergsbaken T, Fink SL, den Hartigh AB et al (2011) Coordinated host responses during pyroptosis: caspase-1–dependent lysosome exocytosis and inflammatory cytokine maturation. J Immunol 187:2748–2754. https://doi.org/10.4049/jimmunol.1100477
Miao EA, Leaf IA, Treuting PM et al (2010) Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria. Nat Immunol 11:1136–1142. https://doi.org/10.1038/ni.1960
Rühl S, Shkarina K, Demarco B et al (2018) ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation. Science 362:956–960. https://doi.org/10.1126/science.aar7607
Toda G, Yamauchi T, Kadowaki T, Ueki K (2020) Preparation and culture of bone marrow-derived macrophages from mice for functional analysis. STAR Protoc 2:100246. https://doi.org/10.1016/j.xpro.2020.100246
Loomis WP, den Hartigh AB, Cookson BT, Fink SL (2019) Diverse small molecules prevent macrophage lysis during pyroptosis. Cell Death Dis 10:326. https://doi.org/10.1038/s41419-019-1559-4
Bauernfeind FG, Horvath G, Stutz A et al (2009) Cutting edge: NF-κB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression. J Immunol 183:787–791. https://doi.org/10.4049/jimmunol.0901363
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Loomis, W.P., Bergsbaken, T. (2023). Monitoring Calcium Fluxes and Lysosome Exocytosis During Pyroptosis. In: Fink, S.L. (eds) Pyroptosis. Methods in Molecular Biology, vol 2641. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3040-2_14
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3040-2_14
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3039-6
Online ISBN: 978-1-0716-3040-2
eBook Packages: Springer Protocols