Comprehensive mRNA profiling of lipid-related genes in microglia and macrophages using taqman arrays.
Quantitative real-time reverse-transcription (RT)-PCR is a precise and sensitive method to measure mRNA levels over a broad dynamic range. This chapter describes the quantitative transcript analysis of 41 selected lipid-related transcripts in macrophages and microglia using a novel "Lipidomic" Taqman Array. The Taqman Array results show that (1) stimulation with the liver-X-receptor and retinoid-X-receptor ligands T0901317 and 9-cis retinoic acid induces several genes of lipid metabolism, (2) lipopolysaccharide (LPS) and interferon-g (Ifn-g) strongly repress lipid-related genes, and (3) coincubation with docosahexaenoic acid dampens the repressing effect of LPS. The method described in this chapter can be used to monitor the transcriptional response of 41 dynamic "lipid" genes simultaneously in any cell type.
Synlab Medical Care Service, Medical Care Center, Weiden, Germany.
Richard
R
Mauerer
Yana
Y
Walczak
Thomas
T
Langmann
D000818:Animals; D002460:Cell Line; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology; D050356:Lipid Metabolism / Q000187:drug effects / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D017628:Microglia / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013449:Sulfonamides / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology; D005466:Fluorocarbons; D000096926:Benzenesulfonamides
D006845:Hydrocarbons, Fluorinated; D007371:Interferon-gamma; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D013449:Sulfonamides; D014212:Tretinoin; C423915:T0901317; D005466:Fluorocarbons; D000096926:Benzenesulfonamides