VDR-dependent signaling modulates neuroinflammation in the acute phase of ischemic stroke

VDR deficiency in microglia markedly aggravated infarct volumes, neurological deficits, and neuroinflammation after cerebral ischemia. To investigate the role of VDR in microglia-regulated neuroinflammation, we performed RNA-seq analysis on microglia isolated from poststroke microglia-conditional VDR knockout (Vdr-cKO) mice and littermate controls. VDR elimination dramatically alters the gene expression profiles of postischemic microglia, implying that vitamin D signaling play a crucial role in microglia-modulated stroke pathogenesis. Brain single-cell suspensions were prepared from Vdr-cKO and Vdrf/f mice 3 days after middle cerebral artery occlusion (MCAO) or sham procedures. Cells were stained with APC-conjugated anti-CD45 and BV421-conjugated anti-CD11b antibodies. CD11b+CD45int microglia were sorted using FACS Aria III flow cytometer for RNA-seq analysis.