Stable isotope labeling with amino acids in cell culture-based proteomic analysis of ethanol-induced protein expression profiles in microglia.
Ethanol exposure causes neurotoxicity, where neuroinflammation has been proposed to contribute to ethanol neurotoxicity. In addition to astroglia, microglia, as resident immune cells in the central nervous system, have been implicated as a key contributor to the neuroimmune and inflammatory processes. However, little is known regarding the role of microglia in alcohol-induced neuronal dysfunction. In this chapter, we describe a method that provides an effective and unbiased global-scale analysis for relative quantitation of protein expression in microglial cells to elucidate the molecular mechanisms underlying microglial activation after ethanol exposure. The approach involves stable isotope labeling with amino acids in cell culture followed by mass spectrometric analysis of stable isotope-labeled proteins derived from cultured microglial cells and represents a powerful tool that can be used for general assessment of microglial response at the protein level.
Department of Pharmacodynamics, University of Florida, Gainesville, FL, USA.
Bin
B
Liu
David S
DS
Barber
Stanley M
SM
Stevens
D000437:Alcoholism / Q000473:pathology; D000596:Amino Acids / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques; D002460:Cell Line; D000431:Ethanol / Q000378:metabolism* / Q000494:pharmacology*; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism; D020258:Neurotoxicity Syndromes / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D040901:Proteomics / Q000379:methods
D000596:Amino Acids; D015415:Biomarkers; D000431:Ethanol