During FY14 we accomplished the following: 1. We initiated studies to examine the response of brain cells to pro-inflammatory cytokines. We started with neuronal cell lines and astrocytomas of human origin. Cells were treated with TNFα, IL-1βor RAGE-ligand for different time periods followed by RNA analysis. The kinetics of NF-κB p65 (RelA) translocation was determined by confocal microscopy, and dynamics of NF-κB binding to target sites is being determined by ChIP-Seq. Cumulatively, we expect that these studies will provide a comprehensive kinetic view of the inflammatory response of neurons and other brain-resident cell types. 2. To independently substantiate the observations in cell lines, we established a collaboration with Dr. Mark Mattsons section (LNS, NIA) to evaluate the responses of primary mouse neurons, microglia and astrocytes to pro-inflammatory cytokines. These studies are in progress.
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