Sexual dimorphism in developmental programming caused by CSF2

项目摘要 植入前胚胎的发育程序因其微环境的改变而改变。 由此产生的发育变化可以产生长期影响，延伸到出生后的生活。不适当 母亲和胚胎之间的信号传递可能会导致更多的流产和长期的健康变化。 对发展规划的研究可以消除与发展相关的不良后果。 与试管婴儿和导致新的方法，以改善人类健康和动物生产普遍。一 胚胎植入前发育程序的一个特点是胚胎发育过程中的性二态性。 成人表型的改变。胚胎对微环境变化反应的性别差异 可能部分由胚胎对母体调节信号的性别特异性反应介导。一 参与这种现象的分子是集落刺激因子2（CSF 2）。在牛中，治疗桑椹胚和 含CSF 2的囊胚期胚胎在妊娠后期（第15天）改变了胚胎特征 以性别特异性的方式，至少在雌性后代中，导致小牛的生长速度增加， 生命的第一年。长期目标是了解胚胎的微环境如何与 性来塑造发展计划。当前提案的总体目标是：1）阐明 CSF2对雄性和雌性胚胎产生差异编程作用的机制，以及2） 表征CSF 2在体外和体内对植入前胚胎的作用的后果， 随后胚胎和出生后表型。假设对CSF 2的性别依赖性反应 依赖于男性和女性胚胎在CSF 2重塑DNA甲基化方面的差异， 这些DNA甲基化的变化会对发育程序产生长期影响， 产后表现型的后果。有三个具体目标。对于目标1，将检验是否 第15天雄性和雌性植入前胚胎在CSF 2编程行为中的差异 胚胎依赖于DNA甲基化。对于目标2，CSF 2对胚胎发育的重要性 通过测试敲除胚胎延长的CSF 2受体的结果来评估体内， 基因表达和DNA甲基化。目标3将侧重于确定 植入前雌性胚胎暴露于CSF 2程序发育以改变出生后表型， 一种提高奶牛生产力的方法。通过这些目标，我们将阐明重塑的重要性 CSF2的性别依赖性作用的甲基化组（目的1），并确定CSF2的作用是否发生 不仅在体外，而且对于在子宫内发育的胚胎（Aim 2）。最后，目标3的实验代表了 这是第一次尝试通过使用发展规划来提高食用动物的生产力， 分子来操纵植入前期的发育。 .

PROJECT SUMMARY The developmental program of the preimplantation embryo is modified by alterations in its microenvironment. The resultant change in development can have long-acting effects that extend into postnatal life. Inappropriate signaling between mother and embryo can lead to increased pregnancy loss and long-term changes in health. Research on developmental programming could lead to elimination of adverse outcomes associated with IVF and result in new approaches to improve human health and animal production generally. One characteristic of developmental programming during the preimplantation period is sexual dimorphism in the modification in adult phenotype. Sex differences in embryonic responses to changes in the microenvironment are likely mediated in part by sex-specific responses of the embryo to maternal regulatory signals. One molecule involved in this phenomenon is colony stimulating factor 2 (CSF2). In cattle, treatment of morula and blastocyst stage embryos with CSF2 changed embryonic characteristics at a later point in pregnancy (Day 15) in a sex-specific manner and, at least in female offspring, resulted in calves with increased growth rates in the first year of life. The long-term goal is to understand how the microenvironment of the embryo interacts with sex to shape the developmental program. The overall objectives of the current proposal are to 1) elucidate mechanisms by which CSF2 causes differential programming actions on male and female embryos and 2) characterize the consequences of actions of CSF2 on the preimplantation embryo in vitro and in vivo for subsequent embryonic and postnatal phenotype. It is hypothesized that sex-dependent responses to CSF2 depend upon differences between male and female embryos in remodeling of DNA methylation by CSF2 and that these changes in DNA methylation result in long-acting effects on the developmental program that have consequences for postnatal phenotype. There are three specific aims. For Aim 1, it will be tested whether differences between male and female preimplantation embryos in programming actions of CSF2 on the Day 15 embryo are dependent upon DNA methylation. For Aim 2, the importance of CSF2 for embryonic development in vivo will be evaluated by testing consequences of knocking out the CSF2 receptor for embryo elongation, gene expression and DNA methylation in male and female embryos. Aim 3 will focus on ascertaining whether exposure of the preimplantation female embryo to CSF2 programs development to alter postnatal phenotype in a manner that improves dairy cow productivity. Through these aims, we will elucidate importance of remodeling of the methylome for sex-dependent actions of CSF2 (Aims 1) and ascertain whether actions of CSF2 occur not only in vitro but also for embryos developing in utero (Aim 2). Finally, the experiment for Aim 3 represents the first attempt to enhance productivity of a food animal through use of a developmental programming molecule to manipulate development in the preimplantation period. .