Project 3: Chromatin and Gene Regulation in Drug Addiction
PI, Eric Nestler (Mount Sinai)
(with Christopher Cowan (UT Southwestern)

All of Projects of this Program Project Grant (PPG) focus on transcriptional mechanisms of cocaine and opiate addiction. Specifically, we focus on two transcription factors, CREB and ?FosB, and selected target genes, within brain reward regions, in particular, the ventral tegmental area (VTA) and nucleus accumbens (NAc), in rodent addiction models. An important component of this work has been the use of DNA expression arrays to study differences in mRNA levels within these brain regions after chronic exposure to a drug of abuse. This has been expanded recently to include arrays for miRNA's (microRNA's), endogenous inhibitors of mRNA expression. This Project oversees this arraying work and analysis for the entire PPG in conjunction with the other Projects. As well, a major advance over the last 3 years of the PPG has been the development of experimental protocols to study changes in chromatin structure within brain reward regions in animal models of drug addiction. This includes chromatin immunoprecipitation (ChIP) assays as well as ChIP combined with high throughput sequencing, so-called "ChIP-Seq" assays. These exciting techniques are now being applied to PPG studies as a major new initiative for our research program. By centralizing all of these activities into this new Project, we ensure a high quality of data and the ready ability to compare and contrast findings across the various Projects. Economic savings are also achieved. We should mention that DNA expression array analyses were included in the Transgenic Core of the currently funded PPG. However, given the greatly expanded use of these arrays, expansion to analysis of miRNA's, our substantial new initiatives in chromatin remodeling, and the fact that this work is now much more investigational (as opposed to serving a core function), it makes sense to create this new Project, which supports highly cohesive studies of chromatin and gene regulation in addiction models.

Distribution of H3K9me3 (trimethylation of histone H3 at Lys 9), a major heterochromatic mark of gene repression, in nucleus accumbens after chronic administration of cocaine using advanced ChIP-Seq methods. From Maze et al., Proc. Natl. Acad. Sci. USA, 2011.

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