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

出生体重和产后健康之间的反向关系是哺乳动物胎儿生物学中最引人入胜的发现之一。支配这种关系的主要假设被称为“胎儿程序化”，即遗传和/或表观遗传变化是胎儿发育变化的基础。了解核受体在调节这些发育事件中的整体作用将为我们更好地理解控制胎儿发育的机制提供帮助。到目前为止，许多已发表的研究都集中在母体侮辱(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。尽管这种方法提供了信息，但它未能通过设计解决转录因子(例如核受体)的更全球作用，以及涉及胎儿基因组重新编程的表观遗传修饰。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识别(CHIP)来检查核受体与其各自靶基因启动子的体内结合的变化来解决这一不足。具体地说，通过购买S-4000超声仪和8样品纳米液滴分光光度计，我们将有能力分别分离和测量胎儿发育期间的染色质变化。利用这个独特的模型，我们可以首次研究核受体和表观遗传标记与靶启动子在体内相互作用的发育变化，这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.