Measurement of Cytosolic Mitochondrial DNA After NLRP3 Inflammasome Activation

  1. Antón, Olga M.
  2. Traba, Javier
Libro:
Methods in Molecular Biology

ISSN: 1064-3745,  1940-6029

ISBN: 9781071621431,  9781071621448

Año de publicación: 2022

Páginas: 117-129

Tipo: Capítulo de Libro

DOI: 10.1007/978-1-0716-2144-8_12 GOOGLE SCHOLAR lock_openAcceso abierto editor

Resumen

The NLRP3 inflammasome, a key component of the innate immune system that mediates caspase-1 activation, which in turn induces cleavage of the pyroptosis executioner gasdermin D and the proinflammatory cytokines IL-1β and IL-18, requires two signals to be activated. First, inflammasome priming is achieved after activation of Toll-like receptors, which leads to NF-κB signaling and transcriptional activation of the genes for NLRP3 and IL-1β. Next, the inflammasome complex is activated by a second signal that induces extrusion of mitochondrial DNA to the cytosol of the cell, which leads to its oligomerization by a not fully understood mechanism. Here we describe a simple method that employs quantitative polymerase chain reaction (qPCR) using SYBR green to measure the presence of mitochondrial DNA (mtDNA) in the cytosol, which can be used to measure cytosolic mtDNA levels after inflammasome activation.

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