Epigenetic regulation of Cdk5 in cognition and emotion

项目总结 负价状态可以在小鼠中使用恐惧条件反射和奖励寻求来建模(一个例子 令人沮丧的非奖励)。这种状态会导致单个基因和基因水平的转录异常 在全基因组范围内；这些变化也是性二态的。人们非常感兴趣的一个基因是细胞周期蛋白依赖性基因。 蛋白激酶5(CDK5)。虽然以前的大多数研究都集中在CDK5蛋白激活和 在压力诱发行为中的信号，我们最近报道可卡因暴露和恐惧条件作用导致 雄性小鼠脑内CDK5的转录调控。这份提案将直接检查 CDK5基因激活与CDK5蛋白活性之间的关系。 此外，有新的证据表明，CDK5相关的组蛋白修饰与药物相关。 恐惧诱导的CDK5表达。目前的建议探索了应激诱导的组蛋白的功能相关性。 利用创新的方法在多个行为范式中对小鼠CDK5基因座进行修改 特定于基因座的表观遗传学编辑。这种方法使我们能够概括基因的内生机制。 表达，避免传统的蛋白质表达产生非生理相关的变化 击倒或过度表达。此外，我们应用细胞类型特异性分析来阐明cdk5在 大脑皮质纹状体-边缘环路中负价状态的调节。这项建议 我将研究恐惧条件作用下CDK5的性别特异性表观遗传调控的直接功能相关性 (目标1)和禁欲期间寻找可卡因(目标2)，使用有针对性的表观遗传编辑。性别-和地区- 将应用特定的磷酸蛋白质组学分析来确定相关的CDK5靶标。精确度识别 应激调节特定基因表达的转录和表观遗传机制对 开发有针对性的抗抑郁药物治疗。

PROJECT SUMMARY Negative valence states can be modeled in mice using fear conditioning and reward seeking (an example of frustrative nonreward). Such states lead to transcriptional aberrations at the level of both individual genes and genome-wide; these changes are also sexually dimorphic. One gene of considerable interest is cyclin-dependent kinase 5 (Cdk5). While most prior studies have focused on the mechanisms of Cdk5 protein activation and signaling in stress-evoked behavior, we recently reported that cocaine exposure and fear conditioning lead to transcriptional regulation of Cdk5 in male mouse brain. This proposal will directly examine the connection between Cdk5 gene activation and Cdk5 protein activity. Furthermore, there is emerging evidence that Cdk5-associated histone modifications correlate with drug-, and fear-induced Cdk5 expression. The current proposal explores the functional relevance of stress-induced histone modifications at the murine Cdk5 locus across multiple behavioral paradigms using the innovative approach of locus-specific epigenetic editing. This approach allows us to recapitulate endogenous mechanisms of gene expression, avoiding non-physiologically relevant changes in protein expression resulting from traditional knockout or overexpression. Furthermore, we apply cell-type specific analyses to elucidate Cdk5 function in brain regions across the corticostriatal-limbic circuitry in the regulation of negative valence states. This proposal will examine the direct functional relevance of sex-specific, epigenetic regulation of Cdk5 by fear conditioning (Aim 1) and cocaine seeking during abstinence (Aim 2) using targeted epigenetic editing. Sex- and region- specific phosphoproteomic profiling will be applied to define relevant Cdk5 targets. Identification of the precise transcriptional and epigenetic mechanisms by which stress regulates specific gene expression is critical for the development of targeted antidepressant treatments.