Characterizing effects of sperm- and oocyte-derived epigenetic factors on early embryonic gene expression and offspring metabolic function

项目摘要/摘要 肥胖等代谢性疾病已成为影响三分之一人口的重大健康风险 在全世界范围内，并可能产生毁灭性的并发症。因此，必须了解 代谢性疾病的易感性，以便制定预防策略。广泛的遗传学研究已经 未能解释这一持续的流行病。然而，父母传递的不仅是遗传信息，还包括 表观遗传因素，可以根据环境刺激进行修改，然后影响基因 表情。如果关于父母环境的信息能够被记录在生殖系中，它有可能被 传播给受精卵并影响后代健康。这一概念在哺乳动物中仍然存在争议， 代表着胚胎学领域的巨大知识鸿沟。实验室之前的工作已经证明 喂食低蛋白饮食的父亲的精子携带tRNA片段和微RNA，可以改变基因表达 在胚胎中。因此，在目标1中，精子来源的RNA将被提纯并显微注射到 单性生殖激活的卵母细胞，或单性生殖，缺乏任何父性遗传内容。 注射的单性生殖的转录图谱将通过单胚胎RNA-Seq获得 描述由此导致的胚胎转录组的变化。表观遗传的母体传播 信息没有被描述到与父方相同的程度。因此，在目标2中，类似的 将利用饮食范例来解决关于母亲饮食的信息是否可以 在卵母细胞中携带，并导致胚胎转录组的变化。体外受精胚胎 来自饲喂低蛋白质、高脂肪或对照饮食的母亲将通过单胚胎RNA-Seq进行测序。 这些胚胎将被转移给养母，以产生成年后代，然后 评估糖耐量和胰岛素抵抗。在这一范例中使用体外受精将 确保在胚胎中观察到的任何变化都来自卵母细胞，而不是在 怀孕了。这项研究的完成将阐明父亲和母亲携带的表观遗传因素的影响 通过胚胎转录组上的配子。 这项工作将在马萨诸塞大学医学院完成，由Dr。 奥利弗·兰多。研究金培训计划包括胚胎技术方面的培训，如显微注射。 免疫荧光染色的谱系标记，以及成年小鼠的代谢表型。 获得科学交流经验的机会包括参加部门研讨会和 地方和国家会议。此外，该大学还在#年举办了职业发展讲习班。 除了对一年级研究生的教学和辅导。

PROJECT SUMMARY/ABSTRACT Metabolic diseases such as obesity have become significant health risks affecting one-third of the population worldwide and can have devastating complications. It is therefore imperative to understand the causes of metabolic disease predisposition in order to develop preventative strategies. Extensive genetic studies have failed to explain this ongoing epidemic. However, parents pass on not only genetic information, but also epigenetic factors, which can be modified in response to environmental stimuli, and can then affect gene expression. If information about parental environment can be recorded in the germline, it has the potential to be transmitted to the zygote and impact offspring health. This concept remains controversial in mammals and represents a large knowledge gap in the field of embryology. Previous work in the lab has demonstrated that sperm from fathers fed a low protein diet carry tRNA fragments and microRNAs that can modify gene expression in the embryo. Therefore, in Aim 1, sperm-derived RNAs will be purified and microinjected into parthenogenetically-activated oocytes, or parthenotes, which lack any paternal genetic content. Transcriptomic profiles of injected parthenotes will be acquired by single-embryo RNA-Seq to characterize resulting alterations to the embryonic transcriptome. Maternal transmission of epigenetic information has not been characterized to the same extent as the paternal side. Therefore, in Aim 2 a similar dietary paradigm will be utilized to address the question of whether information about maternal diet can be carried in oocytes and result in changes to the embryonic transcriptome. In vitro-fertilized embryos from mothers fed low protein, high fat, or control diet will be sequenced by single-embryo RNA-Seq. These embryos will be transferred to foster mothers to produce adult offspring, which will then be assessed for glucose tolerance and insulin resistance. The use of in vitro fertilization in this paradigm will ensure that any changes observed in the embryo originate from the oocyte and not from nutrient exchange during gestation. Completion of this research will elucidate effects of paternal and maternal epigenetic factors carried by gametes on the embryonic transcriptome. This work will be completed at the University of Massachusetts Medical School under the sponsorship of Dr. Oliver Rando. The fellowship training plan includes training in embryological techniques, such as microinjection and immunofluorescent staining of lineage markers, as well as metabolic phenotyping of adult mice. Opportunities to gain experience in science communication include participation in departmental seminars and local and national conferences. Furthermore, career development workshops are provided by the university, in addition to teaching and mentoring of first-year graduate students.