Establishing a tractable epigenetic clock in Nasonia vitripennis

衰老是一个复杂的过程，受众多遗传和环境因素的影响。研究衰老很麻烦，特别是在复杂的哺乳动物模型中。衰老研究主要是使用简单的无脊椎动物模型进行的。这些模式有许多优势。然而，目前的无脊椎动物模型由于缺乏DNA甲基化，在衰老的表观遗传学研究中用处不大。本项目旨在建立一个易处理的表观遗传学时钟，这将有助于形成表观遗传衰老的昆虫模型。纳斯尼亚拥有一个功能强大的DNA甲基化系统，这使它成为这一目的的理想候选者。该项目将使用寿命实验和全基因组亚硫酸氢盐测序(WGBS)来测量药物改变和滞育的Nasonia的时间和表观遗传老化。该项目的目的是：-发现甲基化改变剂地西他滨对表观遗传衰老和寿命的影响。-发现已知的a-酮戊二酸延长寿命的作用是否与减少昆虫的表观遗传老化有关，以便我们能够弥合药物引起的表观遗传时钟变化和实际延长寿命之间的差距。-发现幼虫在生命早期滞育是否会改变成虫的表观遗传老化，以便我们能够了解早期生活史对成虫表观遗传老化的影响。WGBS将被用来计算表观遗传时钟。BS-Seq文库将被准备、测序并与参考基因组比对。为了识别年龄显著的CPG，将进行惩罚性回归。表观遗传年龄将从这个模型中提取出来，并根据年代年龄绘制出来。建立一种易处理的昆虫表观遗传时钟将在生物老年学中开辟一个新的领域，深入研究表观遗传时钟延展性的分子机制，并研究表观遗传衰老的基础生物学。

Ageing is a complex process which is influenced by a myriad of genetic and environmental factors. Investigating ageing is troublesome, particularly in complex mammalian models. Ageing research is largely conducted using simple model invertebrates. There are a number of advantages to these models. However, current invertebrate models are of little use in the study of epigenetics in ageing due to their lack of DNA methylation.This project aims to establish a tractable epigenetic clock in the wasp Nasonia vitripennis, which will aid in the formation of an insect model for epigenetic ageing. Nasonia possess a functional DNA methylation system, making it an ideal candidate for this purpose. This project will use life span experiments and whole genome bisulfite sequencing (WGBS) to measure chronological and epigenetic ageing in drug altered and diapaused Nasonia. The aims of this project are: - To discover the effects of the methylation altering agent decitabine on epigenetic ageing and lifespan. - To discover if the known lifespan increasing effects of a-ketoglutarate are associated with a reduction in epigenetic ageing in insects so that we can bridge the gap between drug induced changes in epigenetic clocks and real increases in lifespan. - To discover if larval diapause early in life alters adult epigenetic ageing so that we can understand the effects of early life history on epigenetic ageing in adult insects. WGBS will be used to calculate the epigenetic clock. BS-Seq libraries will be prepared, sequenced, and aligned to the reference genome. A penalised regression will be conducted in order to identify age significant CpGs. Epigenetic age will be extracted from this model and plotted against chronological age. The establishment of a tractable insect epigenetic clock will open up a new area in biogerontology, delving into the molecular mechanisms of the malleability of the epigenetic clock and investigating the underpinning biology of epigenetic ageing.