Investigation on Lipopolysaccharide Activated Microglia by Phosphoproteomics and Phosphoinositide Lipidomics

Abstract

Microglia are the confined immune cells of the central nervous system (CNS). In response to injury or infection, microglia readily become activated and release proinflammatory mediators that are believed to contribute to microglia-mediated neurodegeneration. In the present study, inflammation was induced in the immortalized murine microglial cell line BV-2 by lipopolysaccharide (LPS) treatment. We firstly performed phosphoproteomics analysis and phosphoinositide lipidomics analysis with LPS activated microglia in order to compare phosphorylation patterns in active and inactive microglia and to detect the pattern of changes in phosphoinositide regulation upon activation of microglia. Mass spectrometry analysis of the phosphoproteome of the LPS treatment group compared to that of the untreated control group revealed a notable increase in the diversity of cellular phosphorylation upon LPS treatment. Additionally, a lipidomics analysis detected significant increases in the amounts of phosphoinositide species in the LPS treatment. This investigation could provide an insight for understanding molecular mechanisms underlying microglia-mediated neurodegenerative diseases.

Optimization of chromatographic buffer conditions for the simultaneous analysis of phosphatidylinositol and phosphatidylinositol phosphate species in canola

“…The similarities between these species may indicate that this second variable fatty acid is one of the most significant sources of variation between the examined cultivars, so far as the PIlipidome is concerned. An ability to differentiate closely related matrices based entirely on their PI-lipidome may serve as a valuable tool to understand the development and progression of certain diseases, specifically due to the variations these pathologies are known to impart on the PI-lipidome [1,3,6,11,12].…”

“…Such lapses in regulation can likewise result in their irregular molecular distribution, permitting analyses to reveal the development of such pathologies. Hence, continual interest in the activity of these molecules motivates their in-depth characterization [1,3,6,11,12]. Unfortunately, conventional lipidomic methods have proven inadequate when considering these species, as their physical and chemical properties are often prohibitive to their analysis [13].…”