Related Experiment Video

Updated: Mar 28, 2026

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
09:04

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells

Published on: May 22, 2014

21.5K

Instant membrane resealing in nlrp3 inflammmasome activation of endothelial cells

Yang Chen1, Ming Yuan, Min Xia

  • 1Department of Pharmacology and Toxicology, School of Medicine, Virginia Commonwealth University, 1220 East Broad Street, Richmond, VA 23298, 2Department of Pharmacological & Pharmaceutical Sciences College of Pharmacy, University of Houston, 3605 Cullen B.

Frontiers in Bioscience (Landmark Edition)
|December 29, 2015
PubMed

Insights

Instant cell membrane resealing (CMR) prevents Nlrp3 inflammasome activation after membrane injury. FasL disrupts CMR, leading to inflammasome activation, but methyl-beta-cyclodextrin (MCD) can block this effect.

Area of Science:

  • Cell Biology
  • Immunology
  • Molecular Mechanisms

Background:

  • Nod-like receptor pyrin domain containing 3 (Nlrp3) inflammasomes are crucial immune sensors.
  • Cell membrane injury can trigger Nlrp3 inflammasome activation.
  • The role of cell membrane resealing (CMR) in regulating inflammasome activation is not fully understood.

Purpose of the Study:

  • To investigate the molecular mechanisms linking instant cell membrane resealing (CMR) to Nlrp3 inflammasome activation.
  • To elucidate the role of membrane rafts (MR) in CMR and subsequent inflammasome responses.
  • To determine how FasL affects CMR and Nlrp3 inflammasome activation.

Main Methods:

  • Utilized wavelength-switching fluorescent microscopy to monitor CMR and intracellular calcium ((Ca(2+))i) in mouse microvascular endothelial cells (MVECs).
  • Induced membrane injury using LCWE or saponin.
  • Investigated the effects of FasL, methyl-beta-cyclodextrin (MCD), VA64, and acid sphingomyelinase siRNA on CMR and inflammasome activation.

Main Results:

  • Membrane injury was resealed in a Ca(2+)-dependent manner, but FasL abolished this process, prolonging resealing time.
  • FasL-induced failure of CMR led to Nlrp3 inflammasome activation, characterized by increased lysosomal permeability and cathepsin B release.
  • Methyl-beta-cyclodextrin (MCD), VA64, and acid sphingomyelinase siRNA blocked FasL-induced inflammasome activation by facilitating CMR.

Conclusions:

  • Membrane raft (MR)-associated CMR is a critical protective mechanism against Nlrp3 inflammasome activation following membrane injury.
  • Disruption of CMR by FasL promotes inflammasome activation.
  • Targeting MR-associated CMR presents a potential therapeutic strategy to control Nlrp3 inflammasome-mediated inflammation.

More Related Videos

Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages06:52

Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages

Published on: May 21, 2018

11.4K
Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages08:41

Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages

Published on: April 6, 2022

3.2K

Related Concept Videos

IP3/DAG Signaling Pathway01:11

IP3/DAG Signaling Pathway

Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and...
15.8K
Intracellular Signaling Affects Focal Adhesions01:17

Intracellular Signaling Affects Focal Adhesions

Integrins act both as extracellular input receivers and as intracellular processing activators. As their name suggests, integrins are entirely integrated into the membrane structure. Their hydrophobic membrane-spanning regions interact with the phospholipid bilayer's hydrophobic region. These membrane receptors provide extracellular attachment sites for effectors like hormones and growth factors. They activate intracellular response cascades when their effectors are bound and active.
Some...
3.8K
Regulation of the Unfolded Protein Response01:31

Regulation of the Unfolded Protein Response

Inositol-requiring kinase one or IRE1 is the most conserved eukaryotic unfolded protein response (UPR) receptor. It is a type I transmembrane protein kinase receptor with a distinctive site-specific RNase activity. As the binding mechanics of the misfolded proteins with the N-terminal domain of IRE-1 are unclear, three binding models — direct, indirect, and allosteric -- are proposed for receptor activation. Nevertheless, it is known that once a misfolded protein associates with IRE1, it...
3.2K
NF-κB-dependent Signaling Pathway02:26

NF-κB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The...
10.3K
Inflammation01:38

Inflammation

Overview
64.2K