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EicosaCell: An Imaging-Based Assay to Identify Spatiotemporal Eicosanoid Synthesis
Christianne Bandeira-Melo1, Ligia Almeida Paiva2,3, Natália R T Amorim2
1Laboratório de Inflamação, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. cbmelo@ufrj.biof.br.
Methods in Molecular Biology (Clifton, N.J.)
|February 11, 2017
Summary
Eicosanoid synthesis compartments can now be detected within cells using the EicosaCell assay. This method immobilizes eicosanoids, enabling visualization of their intracellular production sites.
Area of Science:
- Biochemistry
- Cell Biology
- Lipid Metabolism
Background:
- Eicosanoids are crucial bioactive lipids regulating physiological and pathological processes.
- Their synthesis involves arachidonic acid metabolism via cyclooxygenase (COX) and lipoxygenase (LOX) pathways.
- Understanding intracellular localization of eicosanoid synthesis is key to regulating their function.
Purpose of the Study:
- To introduce and detail the EicosaCell protocol for intracellular eicosanoid detection.
- To enable localization of eicosanoid-synthesizing machinery within cellular compartments.
- To provide a method for studying the functional impact of eicosanoid compartmentalization.
Main Methods:
- The EicosaCell protocol utilizes a strategy to covalently cross-link and immobilize eicosanoids.
- Immunofluorescence techniques are employed to localize the immobilized eicosanoids.
- This allows for the identification of specific intracellular compartments involved in eicosanoid synthesis.
Main Results:
- EicosaCell assays successfully detect and localize eicosanoid synthesis compartments intracellularly.
- The method has been used to identify distinct synthesis sites for prostaglandins and leukotrienes.
- Successful application in cellular activation models demonstrates its utility.
Conclusions:
- The EicosaCell assay provides a powerful tool for visualizing intracellular eicosanoid production.
- It facilitates a deeper understanding of eicosanoid regulation and function.
- This method advances research into lipid mediator signaling pathways.
