Macrophage Necrosis through a Mitochondrial-Lysosomal-ER Circuit DOJINDO LABORATORIES

Previous Science Note

This article focusing on the mechanisms of programmed necrosis in infected macrophages. The research demonstrates that the excess tumor necrosis factor triggers programmed necrosis of infected macrophages is not mitochondrion-intrinsic but results from an inter-organellar circuit initiating and culminating in the mitochondrion. The circuit begins and ends with the transit of two inorganic signals - ROS from mitochondrion to lysosome and Ca²⁺ from endoplasmic reticulum to mitochondrion - and requires cathepsin D translocation from lysosome to cytosol.

TNF Induces Pathogenic Programmed Macrophage Necrosis in Tuberculosis through a Mitochondrial-Lysosomal-Endoplasmic Reticulum Circuit

Click here for the original article: Francisco J. Roca, et. al., Cell (2019)

Point of Interest

- TNF triggers mitochondrial ROS production in mycobacterium-infected macrophages, causing necrosis.
- This process involves mitochondrial ROS activating lysosomal enzymes, leading to BAX activation and subsequent ER activation causing Ca²⁺ influx into mitochondria.
- Targeting this mitochondrial Ca²⁺ overload can help prevent detrimental macrophage necrosis in TB.

Related Techniques

Total ROS detection

Higher sensitivity or for long-term live cell imaging HOT

Mitochondrial superoxide detection

MitoBright ROS - Mitochondrial Superoxide Detection

Glycolysis/Oxidative phosphorylation Assay

Glycolysis/OXPHOS Assay Kit

Glycolysis-related metabolites assay

Glucose and Lactate Assay Kit

Oxygen consumption rate assay

Extracellular OCR Plate Assay Kit

Mitochondrial membrane potential detection

JC-1 MitoMP Detection Kit / MT-1 MitoMP Detection Kit

Mitophagy Detection

Mitophagy Detection Kit and Mtphagy Dye

Lysosomal function assay

Lysosomal pH and mass detection Kit

Autophagy detection

DAPGreen / DAPRed (Autophagosome detection), DALGreen (Autolysosome detection)

Related Applications

1. Simultaneously detection of Lysosomal and Mitochondrial Dysfunction

We tried the simultaneous detection of lysosomal and mitochondrial dysfunction in Hela cells treated with CCCP or Antimycin (AN). CCCP and AN are well-known inducers of mitochondrial ROS regarding loss of mitochondrial membrane potential. Recent research showed the result that CCCP induces not only mitochondrial ROS but also lysosomal neutralization. To detect mitochondrial ROS, HeLa cells were labeled by MitoBright ROS - Mitochondrial Superoxide Detection, and the lysosomal mass and pH were detected separately with LysoPrime Green and pHLys Red. Co-staining with mitoBright ROS and Lysosomal dyes demonstrated that CCCP causes lysosomal neutralization and mitochondrial ROS induction at the same time.

Products in Use
- LysoPrime Green
- pHLys Red
- MitoBright ROS - Mitochondrial Superoxide Detection

Related Product
- Lysosomal Acidic pH Detection Kit

2. Monitoring ROS in Macrophage Phagocytosis

Dead cells (2&3) phagocytosed by Cell1 resulted in increased ROS(green).

ROS detection reagent allowed for reliable analysis of the role of ROS in phagocytosis. Its high intracellular residence and low background noise made it possible to perform long-term analysis of ROS production in the cell. This information can provide important insights into the mechanisms of phagocytosis and contribute to the development of treatments for diseases associated with macrophage dysfunction.

> for detail experimental notes are available at Nikon web site.

Products in Use
- ROS Assay Kit -Photo-oxidation Resistant DCFH-DA-

Related Product
- ROS Assay Kit -Highly Sensitive DCFH-DA-