Quantitative Proteomics Reveals Temporal Proteomic Changes in Signaling Pathways during BV2 Mouse Microglial Cell...

Area of Science:

• Systems biology
• Proteomics
• Neuroinflammation

Background:

• Microglia play a crucial role in brain homeostasis and immune responses.
• Microglial activation, particularly the proinflammatory state induced by lipopolysaccharide (LPS) and interferon-gamma (IFN-γ), is implicated in neurodegenerative diseases.
• Understanding proteomic changes during microglial activation is essential for identifying disease mechanisms.

Purpose of the Study:

• To develop and apply a systematic proteomic quantification platform for in-depth analysis of cellular systems.
• To investigate the intracellular and extracellular proteome changes in activated microglia.
• To identify pathways associated with microglial activation and their potential role in neurodegeneration.

Main Methods:

• Utilized an improved shotgun proteomics approach for global proteome analysis.
• Employed label-free parallel reaction monitoring (PRM) for high-resolution mass spectrometry-based targeted quantification.
• Applied a systematic proteomic platform encompassing global to targeted analysis on a single platform.

Main Results:

• Identified 5497 proteins in the whole-cell proteome and 4938 proteins in the secretome of LPS/IFN-γ-stimulated BV2 mouse microglia.
• Classified differentially expressed proteins into pathways related to immune-inflammatory responses and metabolism.
• Quantified over 450 peptides corresponding to pathway proteins and interactors using label-free PRM in a single run.

Conclusions:

• The developed systematic quantification platform enables comprehensive analysis of activated microglia proteomes.
• Data suggest latent pathways associated with neurodegenerative diseases can be discovered through pathway network modeling.
• The platform holds significant potential for large-scale targeted proteomic analyses in various biological systems.