Identification and functional characterization of ATF3 target genes that mediate neuronal survival

神经元的活性和NMDA受体的激活对神经元的存活至关重要。虽然这一过程的确切机制尚不清楚，但由转录因子CREB介导的活性诱导的核钙瞬变和基因表达是重要的。全基因组转录分析导致了核钙调控，多组分基因组生存程序的发现。CREB靶点ATF3在该计划中起核心作用；它作为一种转录抑制因子，通过下调基因来提供神经保护。该项目的目的是确定介导神经保护的ATF3靶点。我们提出了两种实验策略：对ATF3靶基因进行无偏搜索，以及对假定的ATF3靶基因Trpm7进行分析，Trpm7是一种与细胞死亡有关的非选择性阳离子通道。我们将使用全基因组转录组分析来鉴定海马神经元中所有受ATF3调控的基因，并使用基因本体搜索来过滤掉那些可能在细胞凋亡或存活中起作用的基因。Trpm7在海马神经元电激活时的表达遵循与它是ATF3靶标一致的表达谱。我们将研究核钙- creb - atf3信号对所有候选靶基因（包括Trpm7）的调控，并评估它们在体外和体内神经元存活中的作用。这项研究可能为神经退行性疾病的神经保护疗法的开发提供新的基因靶点。

Neuronal activity and NMDA receptor activation are critical for the survival of neurons. Although the precise mechanisms involved in this process are unknown, activity-induced nuclear calcium transients and gene expression mediated by the transcription factor CREB are important. Whole genome transcriptional profiling led to the discovery of a nuclear calcium-regulated, multi-component genomic survival program. The CREB target ATF3 plays a central role in this program; it acts as a transcriptional repressor and confers neuroprotection through down-regulation of genes. The aim of the project is to identify the ATF3 targets that mediate neuroprotection. We propose two experimental strategies: an unbiased search for ATF3 target genes as well as an analysis of the putative ATF3 target gene, Trpm7, a non-selective cation channel with a role in cell death. We will use whole genome transcriptome analysis to identify all genes regulated by ATF3 in hippocampal neurons and, using gene ontology searches, filter out those genes that have a possible role in apoptosis or survival. Expression of Trpm7 upon electrical activation of hippocampal neurons follows an expression profile consistent with it being a target of ATF3. We will investigate the regulation of all candidate target genes (including Trpm7) by nuclear calcium-CREB-ATF3 signaling and assess their role in neuronal survival in vitro and in vivo. This study may lead to the identification of novel gene targets for the development of neuroprotective therapies for neurodegenerative diseases.