Transcriptome analysis of alcohol-treated microglia reveals downregulation of beta amyloid phagocytosis

• Published in: Journal of neuroinflammation 2018-05, Vol.15 (1), p.141-11, Article 141
• Main Authors: Kalinin, Sergey, González-Prieto, Marta, Scheiblich, Hannah, Lisi, Lucia, Kusumo, Handojo, Heneka, Michael T, Madrigal, Jose L M, Pandey, Subhash C, Feinstein, Douglas L
• Format: Article
• Language: English
• Subjects: Alcohol; Alcoholic beverages; Alzheimer’s disease; Amyloid; Amyloid beta-Peptides - antagonists & inhibitors; Amyloid beta-Peptides - metabolism; Amyloid beta-protein; Animals; Animals, Newborn; Cells, Cultured; Complement; Down-Regulation - drug effects; Down-Regulation - physiology; Ethanol - toxicity; Female; Gene Expression Profiling - methods; Health aspects; Male; Microglia; Microglia - drug effects; Microglia - metabolism; Peptide Fragments - antagonists & inhibitors; Peptide Fragments - metabolism; Phagocytosis; Phagocytosis - drug effects; Phagocytosis - physiology; Rats; Rats, Sprague-Dawley; RNA, Messenger - antagonists & inhibitors; RNA, Messenger - metabolism; Transcription (Genetics)
• ISSN: 1742-2094; 1742-2094
• DOI: 10.1186/s12974-018-1184-7

Microglial activation contributes to the neuropathology associated with chronic alcohol exposure and withdrawal, including the expression of inflammatory and anti-inflammatory genes. In the current study, we examined the transcriptome of primary rat microglial cells following incubation with alcohol alone, or alcohol together with a robust inflammatory stimulus. Primary microglia were prepared from mixed rat glial cultures. Cells were incubated with 75 mM ethanol alone or with proinflammatory cytokines ("TII": IL1β, IFNγ, and TNFα). Isolated mRNA was used for RNAseq analysis and qPCR. Effects of alcohol on phagocytosis were determined by uptake of oligomeric amyloid beta. Alcohol induced nitrite production in control cells and increased nitrite production in cells co-treated with TII. RNAseq analysis of microglia exposed for 24 h to alcohol identified 312 differentially expressed mRNAs ("Alc-DEs"), with changes confirmed by qPCR analysis. Gene ontology analysis identified phagosome as one of the highest-ranking KEGG pathways including transcripts regulating phagocytosis. Alcohol also increased several complement-related mRNAs that have roles in phagocytosis, including C1qa, b, and c; C3; and C3aR1. RNAseq analysis identified over 3000 differentially expressed mRNAs in microglia following overnight incubation with TII; and comparison to the group of Alc-DEs revealed 87 mRNAs modulated by alcohol but not by TII, including C1qa, b, and c. Consistent with observed changes in phagocytosis-related mRNAs, the uptake of amyloid beta , by primary microglia, was reduced by alcohol. Our results define alterations that occur to microglial gene expression following alcohol exposure and suggest that alcohol effects on phagocytosis could contribute to the development of Alzheimer's disease.