Predoctoral Fellowship for Amanda Prewitt: Estrogen Receptor Alpha mRNA Regulation in the Mouse Cortex Prior to and Following a Stroke-Like Injury

This project will address several issues. It will characterize the promoters responsible for ER-a mRNA expression during mouse cortical development utilizing rapid amplification of cDNA ends (RACE) will be performed at various time points early in development, to determine the specific ER-a promoter responsible for ER-a mRNA expression and quantitative RT-PCR will determine their usage. This project will also investigate the regulation of ER-a mRNA in an in vitro stroke-like injury utilizing organotypic explant cultures of the cortex and examine ER-a expression prior to and following an in vitro stroke-like injury by Quantitative RT-PCR. We will also use promoter-specific primers to determine which ER-a promoters are utilized. The role methylation of promoters in ER-a mRNA expression after an ischemic event will be examined by organotypic explant cultures and examine and perform methylation specific PCR to determine methylation status of the ER-a promoters prior to and following an in vitro stroke injury. The outcomes of these studies will add to our understanding of how ER-a mRNA is regulated the mouse cortex. An understanding of the regulation of ER-a expression throughout the cortex is not only important to the understanding of basic developmental biology, but also has potential future applications in neuroprotection in brain injury models like stroke.