Epigenetic regulatory mechanisms during neurodevelopment

神经发育过程中的表观遗传调控机制

• 批准号: RGPIN-2016-05531
• 负责人: Singh, Shiva
• 金额: $ 3.21万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=665298
• 关键词: Epigenetic; regulatory; mechanisms; during; neurodevelopment

Among other insights, this NSERC funded research has established that the natural variation associated with gene expression is much more common than DNA polymorphism. From evolutionary perspective, this variation offers among the first defence to environmental stresses particularly during neurodevelopment. Further, we have demonstrated that advances in molecular technologies to appraise this variability reliably now offers effective strategies to investigate this defence during neurodevelopment, which has remained enigmatic. Our strategy is to interrupt neurodevelopment at desired time(s) by stressors (eg. alcohol) and evaluate genome-wide changes using RNA-seq and microarrays. The results have allowed us to conclude that alcohol yields a lifelong “molecular footprint” of its encounter during neurodevelopment. Further, this “footprint” may underlie critical behavioral defects often associated with neurodevelopmental exposure to alcohol. This proposal will specifically focus on molecular studies on hippocampus that directs learning and memory often affected by prenatal alcohol. ***Objective 1: To assess hippocampal gene expression and DNA methylation in the adult mice that show behavioral abnormalities including learning/memory deficits caused by neurodevelopmental alcohol exposure.***Objective 2: To assess if prenatal-alcohol-exposure induced behavioral and molecular landscapes could be altered (improved or worsened) by postnatal environmental enrichment (positive) and maternal separation (negative), respectively.****This research will use C57BL/6J mice, as the animal model. It begins with generation of two groups of time-mated (pregnant) females. The first group is given a choice between 10% ethanol and water as the free choice of liquid (E) and the second group receives only water bottle (W). Pregnant C57BL/6J females prefer (70%) to drink 10% alcohol and the resulting progeny shows multiple behavioral effects including learning and memory deficits. Next, the resulting progeny (21 day old) from E and W mothers are subjected to three postnatal environments; raised with free access to interactive toys and running wheels (enrichment or positive environment, P); subject to maternal separation stress (negative environment, N) and normal rearing (control, C). The animals representing six groups (EP, EN, EC, WP, WN, WC) will be followed for learning and memory using Barnes Maze. The hippocampi of such mice will be assessed for molecular footprint using appropriate technologies. ****The results will offer a novel insight in the molecular interplays between DNA methylation and prenatal (alcohol) and postnatal environment in progression of learning and memory. The project as outlined will train HQPs with expertise in neuroscience, epigenetics and modern experimental protocols involving genomics and epigenomics. Such HQPs are needed in this, the post genome era.**

在其他见解中，这项由NSERC资助的研究已经确定，与基因表达相关的自然变异比DNA多态要常见得多。从进化论的角度来看，这种变异为环境压力提供了第一道防线，尤其是在神经发育期间。此外，我们已经证明，可靠地评估这种可变性的分子技术的进步现在提供了有效的策略来研究神经发育期间的这种防御，这仍然是一个谜。我们的策略是在期望的时间(S)通过应激源(例如。酒精)，并使用RNA-SEQ和微阵列评估全基因组的变化。这些结果让我们得出结论，酒精在神经发育过程中会产生终生的“分子足迹”。此外，这种“足迹”可能是神经发育过程中经常与酒精暴露相关的严重行为缺陷的基础。这项建议将特别关注海马体上指导学习和记忆的分子研究，这些海马体通常会受到产前酒精的影响。*目的1：评估酒精暴露引起的行为异常(包括学习/记忆障碍)成年小鼠的海马区基因表达和DNA甲基化。*目的2：评估出生前酒精暴露诱导的行为和分子景观是否会因出生后环境丰富(阳性)和母体分离(阴性)而改变(改善或恶化)。*本研究将以C57BL/6J小鼠为动物模型。它开始于产生两组时间交配(怀孕)的雌性动物。第一组在10%乙醇和水之间自由选择液体(E)，第二组只接受水瓶(W)。怀孕的C57BL/6J雌性(70%)喜欢喝10%的酒精，由此产生的后代表现出包括学习和记忆障碍在内的多种行为影响。接下来，E和W母亲的后代(21日龄)受到三种出生后环境的影响：自由接触互动玩具和跑轮(丰富或积极环境，P)；母亲分离压力(消极环境，N)和正常养育(对照组，C)。采用Barnes迷宫对EP、EN、EC、WP、WN、WC六组动物进行学习记忆实验。将使用适当的技术评估这些小鼠的海马区的分子足迹。*这些结果将为DNA甲基化与产前(酒精)和出生后环境在学习和记忆进展中的分子相互作用提供新的见解。正如所概述的那样，该项目将培训具有神经科学、表观遗传学和涉及基因组学和表观基因组学的现代实验方案的专家。这种HQP在这个后基因组时代是需要的。**