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Time Resolved IRMS Study of Staurosporine Induced Apoptosis in Murine Astrocyte Cells
Author(s) -
Weber Kayla N,
Husher Sara C,
Mazzer Paula A
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.945.3
Subject(s) - staurosporine , metabolomics , lipidomics , lipid metabolism , necroptosis , apoptosis , biology , proteomics , microbiology and biotechnology , chemistry , biochemistry , computational biology , programmed cell death , bioinformatics , signal transduction , protein kinase c , gene
The advent of genomic and post genomic technologies is transforming the field of toxicology, and is moving the discipline from the realm of classical biology into those of molecular biology and bioanalytical chemistry. At the molecular level, it is possible to focus on both the direct impact of toxic insult as well as the cellular or organismic response to the resulting damage. The “‐omics” technologies—genomics, proteomics, and metabolomics—are likely to lead the way in the development of approaches to molecular pathology. Metabolomics, in fact, shows such promise in understanding the biochemical functioning of cells that the development of metabolomic technologies has been a major NIH roadmap initiative. One subspecialty of metabolomics, lipidomics, shows a special promise because lipids are simple to analyze and have short response time to toxic insult. One shortcoming of current lipidomic exploration is a lack of knowledge of which lipid synthetic pathways are activated in response to toxic insult. Therefore, we undertook to study lipid synthesis in response to apoptotic cell death in murine (C8‐D1A ATCC) astrocyte cells. To investigate time resolved lipid expression we used a model of staurosporine‐induced apoptosis in murine astrocyte cells. We extracted lipids at defined time points after staurosporine insult using a modified Bligh and Dyer technique. Lipids were then sorted by lipid class using rapid HPTLC and then derivatized to fatty acid methyl esters (FAMEs). Time dependent changes in fatty acid expression were observed over the first six hours following staurosporine treatment. Inclusion of 13 C‐labeled glucose immediately prior to staurosporine addition allowed for investigation of de novo fatty acid synthesis by Isotope Ratio Mass Spectroscopy (IRMS). The ability to use IRMS to trace formation of lipid synthesis has the potential to add a new dimension to future lipidomic studies. Support or Funding Information This work was sponsored by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103443.