Measuring changes in nuclear receptor binding during fetal and neonatal development using chromatin immunoprecipitation (ChIP)

出生体重与产后健康之间的负相关关系是哺乳动物胎儿生物学中最令人着迷的发现之一。控制这种关系的主要假设被称为“胎儿编程”，即遗传和/或表观遗传变化是胎儿发育改变的基础。了解核受体在这些发育事件中的整体作用将使我们更好地了解控制胎儿发育的机制。迄今为止，许多已发表的研究都集中在母体损伤（即营养不良和低氧血症）对胎儿和新生儿特定靶基因和生理参数的影响上。尽管这种方法提供了信息，但它未能通过设计来解决转录因子（例如核受体）和表观遗传修饰的更全局作用，这些修饰涉及胎儿基因组的“重编程”。总的来说，本提案中要求的设备将通过促进在组织中使用染色质免疫沉淀（ChIP）来检查核受体与其各自靶基因启动子结合的体内变化来解决这一缺陷。具体来说，随着S-4000超声仪和8样品纳米滴分光光度计的购买，我们将有能力分离和测量染色质变化，分别在胎儿发育过程中。通过这一独特的模型，我们首次研究了核受体和表观遗传标记与靶启动子相互作用的体内发育变化，这些启动子在正常和异常胎儿发育过程中对基因转录的协调控制起着重要作用。

The inverse relationship between birth weight and post partum health is one of the most fascinating discoveries in mammalian fetal biology. The principal hypothesis governing this relationship, termed 'fetal programming', is that genetic and/or epigenetic changes underlie alterations in fetal development. Understanding the overall role of nuclear receptors in mediating these developmental events would provide us with a better understanding of the mechanisms controlling fetal development. To date, many published studies have been focused upon the effect of maternal insults (i.e. malnutrition and hypoxemia) on specific target genes and physiological parameters in the fetus and neonate. Although this approach is informative, it fails to address by design the more global role of transcription factors (e.g. nuclear receptors) and epigenetic modifications involved underlying the 'reprogramming' of the fetal genome. Collectively, the equipment requested in this proposal will address this deficiency by facilitating the use chromatin immunoprecitation (ChIP) in tissues to examine the alterations in vivo binding of nuclear receptors to the promoters of their respective target genes. Specifically, with the purchase of an S-4000 Sonicator, and an 8-sample Nanodrop Spectrophotometer, we will have the ability to isolate and measure chromatin changes, respectively, during fetal development. With this unique model, we can examine for the first time the developmental changes in the in vivo interactions of nuclear receptors and epigenetic markers to target promoters, which play an important role in the coordinated control of gene transcription during normal and abnormal fetal development.