[en] [en] BACKGROUND: 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.
METHODS: 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.
RESULTS: 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 beta1-42, by primary microglia, was reduced by alcohol.
CONCLUSIONS: 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.
Disciplines :
Neurology
Author, co-author :
Kalinin, Sergey; Department of Anesthesiology, University of Illinois at Chicago, Chicago, IL, 60612, USA
González-Prieto, Marta; Department of Pharmacology, University Complutense, Centro de Investigacion Biomedica en Red de Salud Mental (CIBERSAM), Madrid, 28040, Spain
Scheiblich, Hannah; Department of Neurodegenerative Disease and Geriatric Psychiatry, University of Bonn, 53127, Bonn, Germany
Lisi, Lucia; Institute of Pharmacology, Catholic University Medical School, 00168, Rome, Italy
Kusumo, Handojo; Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, Chicago, IL, 60612, USA
HENEKA, Michael ; Department of Neurodegenerative Disease and Geriatric Psychiatry, University of Bonn, 53127, Bonn, Germany
Madrigal, Jose L M; Department of Pharmacology, University Complutense, Centro de Investigacion Biomedica en Red de Salud Mental (CIBERSAM), Madrid, 28040, Spain
Pandey, Subhash C; Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, Chicago, IL, 60612, USA ; Department of Veterans Affairs, Jesse Brown VA Medical Center, Chicago, IL, 60612, USA
Feinstein, Douglas L; Department of Anesthesiology, University of Illinois at Chicago, Chicago, IL, 60612, USA. dlfeins@uic.edu ; Department of Veterans Affairs, Jesse Brown VA Medical Center, Chicago, IL, 60612, USA. dlfeins@uic.edu
External co-authors :
yes
Language :
English
Title :
Transcriptome analysis of alcohol-treated microglia reveals downregulation of beta amyloid phagocytosis.
National Institute on Alcohol Abuse and Alcoholism U.S. Department of Veterans Affairs
Funding text :
This work was supported in part by grant NIAAA P50AA-022538 (SCP) and a Research Career Scientist award from the Department of Veterans Affairs (DLF and SCP).
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