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

paper Cited Public

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
Year: 2018
Journal: Journal of neuroinflammation
PMID: 29759078
DOI: 10.1186/s12974-018-1184-7
PMCID: PMC5952855

Fig. 1

Alcohol increases microglial inflammatory activation. Primary rat microglia were incubated overnight with nothing (control), with TII to induce an inflammatory response, with 75 mM ethanol (Alc), or with TII and ethanol (Alc and TII). The next day, nitrite production was measured using Griess reagent. The data is the mean ± sem of n = 6–15 replicates done in 3 independent studies. *P < 0.05; ***P < 0.0001; one-way ANOVA, Tukey’s post hoc analysis

Fig. 2

Validation of RNAseq results. Reverse transcriptase quantitative PCR (qPCR) was used to measure mRNA levels of the indicated Alc-DEs. qPCR was carried out in samples from microglia incubated for 24 h with nothing (control, n = 3) or 75 mM ethanol (n = 3), and results normalized to values measured for β-actin in the same samples. The y-axis shows the ratio of the average mRNA level measured in the ethanol versus the control samples (filled bars) and is plotted next to the fold-difference calculated from RNAseq data (open bars). For qPCR results, *P < 0.05; #P < 0.10 control vs alcohol. For the RNAseq data, all DEs were found significantly different using Deseq2

Fig. 3

Overlap of identified DEs. A Venn diagram illustrating overlap of Alc-DEs (n = 312; orange, regions 1, 2, 4, and 5), TII-DEs (n = 3082; green, regions 2, 3, 5, and 6) and AlcTII-DEs (n = 3552; blue, regions 4, 5, 6, and 7). The full list of the DEs in each group is provided in Additional file 1: Tables S1-S4

Fig. 4

Alcohol reduces amyloid beta phagocytosis in rat microglia. Primary rat microglia were incubated overnight with a nothing (control), b 75 mM ethanol, c TII, or d 75 mM ethanol and TII. The next day, the cells were assayed by flow cytometry for phagocytosis of FAM-labeled oligomeric Aβ1–42. e Average internalized FAM fluorescence per cell. Data is from 3 different batches of microglial cells totaling 9197 (controls), 8976 (alcohol), 24,588 (TII), and 28,773 (Alc * TII) events. *P < 0.005 versus control cells, one-way ANOVA, Tukey’s post hoc analysis

#	Section	Preview
20	Results — Alcohol reduces microglial phagocytosis	Since phagocytosis was one of the highest KEGG pathways identified in the Alc-DE group, we tested if…
21	Discussion	Several studies have examined the effects of alcohol consumption on neuroinflammatory gene…
22	Discussion	[24], 1010 Alc-DEs were identified in the microglial samples, compared to 2461 in total homogenates.…
23	Discussion	In our study, the average change in the 313 Alc-DEs was 16%, ranging from a 50% decrease to a 72%…
24	Discussion	In addition to identification of genes altered by alcohol use, studies have been carried out in…
25	Discussion	In our studies, pathway analysis using DAVID [41] and GO Consortium [42, 43] platforms identified 5…
26	Discussion	the brain could contribute to alcohol-induced neuropathology. While this may be mediated through…
27	Discussion	a1, also referred to as Nramp1Transmembrane phagosomal divalent cation transporter [57]Mfge8Ligand…
28	Discussion	Our results demonstrate that microglial phagocytosis of Aβ1–42 is significantly suppressed…
29	Discussion	development. It was also shown using a similar acute exposure model, that although activated…
30	Discussion	In our studies, alcohol exposure reduced phagocytosis of Aβ with no effect on uptake of polystyrene…
31	Discussion	Alcohol consumption is generally considered a risk factor for dementia, although there are some…
32	Discussion	3 years found that light to moderate alcohol consumption reduced the risk of overall dementia by…
33	Discussion	In contrast, evidence that alcohol can increase amyloid levels comes from several studies. In vitro,…
34	Discussion	the hippocampus [34]. Long-term alcohol consumption significantly impaired spatial memory in adult…
35	Discussion	The current findings have several limitations, a primary one being that these studies were done…
36	Conclusion	These results define the changes that occur to microglial cells following exposure to…
37	Additional file	Additional file 1:Table S1. Differentially expressed mRNAs due to alcohol treatment “Alc-DEs”.…

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Analysis of whole genome-transcriptomic organization in brain to identify genes associated with alcoholism.	2019	30765688

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