Protective effects of ovarian hormones: sex-specific differences in tlr4 expression after adolescent alcohol

项目摘要/摘要 早期饮酒与酒精使用障碍(AUD)的终生风险增加相关，并且 依赖前额叶皮质的认知功能损害(Jennison，2004；Townshend和Duka， 2005)。了解酒精对发育中的青少年大脑的影响可以帮助我们识别分子和 靶向治疗干预的细胞通路。我们的实验室已经证明，仅仅两周的自愿 青春期早期饮酒(青春期成熟期)导致有髓纤维密度降低 雄性大鼠的前额叶皮质(Vargas等人，2014)，雌性大鼠没有可测量的变化(未发表)。 这种性别差异可能反映了Toll样受体4(TLR4)的差异上调，就像这个受体 显示了协调神经炎性反应(Blanco等人，2010)和介导髓鞘损伤后 重度或长期酒精暴露(Alfonso-Loeches等人，2012年)。为了支持这一假设，我的 学位论文研究证实，饮酒可上调大鼠前额叶皮质TLR4基因的表达 雄性，但不是雌性青春期大鼠(目标1)。性腺激素已被证明具有神经保护作用。 和抗炎作用(Vegeto等人，2003)，然而，它们在酒精介导的干扰中的作用，如 Toll样受体的上调仍不清楚。因此，在本提案的F99博士前阶段，我 将测试循环性腺激素在保护女性免受这些影响方面的作用，并确定 牵涉到神经胶质细胞。在我完成我的论文后，我在K00阶段的目标是研究 青春期酒精暴露后神经免疫系统的适应，以及这些神经适应是如何 可能会在以后的生活中导致酗酒和酒精使用障碍。具体来说，我将使用RNAseq， 转基因动物模型、原代细胞培养和芯片，以及在 F99阶段，回答酒精如何引起表观遗传变化，特别是靶向 神经炎性基因，促进不同类型细胞生物学功能的改变，以及这又是如何 规范行为。我这项研究的最终目标是深入了解酒精的表观遗传学目标， 这将有助于确定治疗药物成瘾和神经退行性疾病的潜在疗法。

PROJECT SUMMARY/ABSTRACT Early onset of alcohol drinking is associated with an increased lifetime risk of alcohol use disorder (AUD) and impairment of cognitive functions dependent on the prefrontal cortex (Jennison, 2004; Townshend and Duka, 2005). Understanding how alcohol impacts the developing adolescent brain can help us identify molecular and cellular pathways to target for therapeutic intervention. Our lab has shown that just two weeks of voluntary alcohol drinking in early adolescence (during pubertal maturation) leads to reduced myelinated fiber density in the prefrontal cortex in males (Vargas et al., 2014), without measurable changes in female rats (unpublished). This sex difference may reflect differential upregulation of toll-like receptor 4 (TLR4), as this receptor has been shown to orchestrate a neuroinflammatory response (Blanco et al., 2010) and mediate myelin damage after heavy or prolonged alcohol exposure (Alfonso-Loeches et al., 2012). In support of this hypothesis, my dissertation research has confirmed that alcohol drinking upregulates TLR4 mRNA in the prefrontal cortex of male, but not female adolescent rats (Aim 1). Gonadal hormones have been shown to have neuroprotective and anti-inflammatory effects (Vegeto et al., 2003), however, their role in alcohol-mediated disruptions, like upregulation of toll-like receptors, remains unknown. Therefore, in the F99 predoctoral phase of this proposal I will test the role of circulating gonadal hormones in protecting females against these effects and identify the glial population involved. After I am done with my dissertation, my goal for the K00 phase is to study how the neuroimmune system adapts after alcohol exposure during adolescence, and how these neuroadaptations may contribute to alcohol abuse and alcohol use disorder later in life. Specifically, I will use RNAseq, transgenic animal models, primary cell cultures, and ChIP, along with mRNA and protein assays learned in the F99 phase, to answer how epigenetic changes induced by alcohol, especially those targeting neuroinflammatory genes, contribute to changes in biological function of different cell types and how this in turn regulates behavior. My ultimate goal for this research is to gain insight into the epigenetic targets of alcohol, which will help identify potential therapeutics for drug addiction and neurodegenerative disorders.