Next-generation RNA sequencing of Acetate treated primary cultured mouse astrocytes in the presence or absence of Aβ

Reactive astrogliosis is a well-known and recognised marker of Alzheimer's Disease (AD). The aim of this study is to functionally visualise reactive astrocyte-mediated neuronal hypometabolism in the brains that is associated with AD and neuroinflammation. To investigate alterations of acetate and glucose metabolism and the effect thereof in AD-like astrocytes, primary astrocyte cultures were treated with acetate in the presence or absence of amyloid beta (Aβ) oligomers. On comparison, we found that acetate treatment upregulated the expression of acetate transporter MCT1 (Slc16a1) as well as that of enzymes involved in the urea cycle and subsequent GABA synthesis. We could, therefore, conclude that MCT1-mediated uptake of acetate into astrocytes triggered GABA synthesis by upregulating enzymes involved in putrescine production and degradation. Primary cultured astrocytes (P0-P1) were treated with Aβ oligomers (1μM) for 5 days (DIV8-12). On the 4th day (DIV11), culture media was replaced with glucose-free medium and supplemented with sodium acetate (10mM). The next day, RNA was isolated from the cultured cells and processed to prepare cDNA libraries compatible with Illumina HiSeq NEXT Generation high-throughput sequencer. After the 2 x 100bp Illumina HiSeq paired-end sequencing run was complete, data in the form of BCL files was procured and processed to obtain relevant findings.