Mitochondrial mechanisms of maternal age effects on offspring health and lifespan

项目总结 生育时孕妇的高龄降低了后代的寿命和健康 包括人类在内的一系列物种。控制母体年龄影响的机制 然而，后代是未知的。在我们的母公司R01项目中，我们假设线粒体 高龄产妇的功能障碍导致功能障碍的线粒体在 通过补偿性生物发生产生后代，减少发育过程中的自噬。这 破坏后代线粒体功能和线粒体-核通讯，导致 加速了后代的衰老。父项目的目标是了解 母体年龄决定后代衰老的线粒体机制。在父级中 R01，我们建议确定引发母体年龄的母体线粒体机制 影响；确定导致线粒体DNA积累和损伤的发育机制 老年母亲后代中的线粒体；并确定后代线粒体的机制 并确定这些机制是否是ROS- 依附的。当提交父项目建议书时，一种CRISPR基因的方法 对臂尾轮虫的编辑仍在开发中。我们现在已经成功地创建了一个 CRISPR介导的臂尾轮虫高效诱变方案。AS 对于这种新方法，我们创建了Vasa和MLH3的敲除突变，并 MLH3中停止盒的敲入突变。突变会遗传给很多人 世世代代，展示了我们建立稳定突变系的能力。是否可以使用 可遗传的CRISPR诱导的轮虫突变现在使检测成为可能 线粒体机制对母体年龄的影响更为直接。这个小规模项目的目标是， 短期行政补充项目是建立5-7个稳定的基因敲除突变体和 马爪哇臂尾轮虫线粒体相关基因的敲入荧光报告菌株。 建立与线粒体功能和动力学相关的基因突变将使我们能够 检测特定线粒体动力学和代谢途径在母亲年龄效应中的作用 亲本R01的寿命和健康寿命及未来研究。

PROJECT SUMMARY Advanced maternal age at the time of reproduction decreases offspring lifespan and health in a range of species, including humans. The mechanisms controlling maternal age effects on offspring are unknown, however. In our parent R01 project, we hypothesize that mitochondrial dysfunction in advanced maternal age causes accumulation of dysfunctional mitochondria in offspring through compensatory biogenesis and decreased autophagy during development. This disrupts offspring mitochondrial function and mitochondrial-nuclear communication, leading to accelerated offspring aging. The objectives of the parent project are to understand the mitochondrial mechanisms by which maternal age determines offspring aging. In the parent R01, we proposed to identify the maternal mitochondrial mechanisms that trigger maternal age effects; identify the developmental mechanisms causing accumulation of mtDNA and damaged mitochondria in old-mother offspring; and identify the offspring mitochondrial mechanisms involved in negative maternal age effects and determine if these mechanisms are ROS- dependent. When the parent project proposal was submitted, a method for CRISPR gene editing in Brachionus rotifers was still in development. We have now successfully created a protocol for high-efficiency CRISPR-mediated mutagenesis in Brachionus manjavacas. As proof-of-principle for this new method, we created knockout mutations of VASA and MLH3, and a knock-in mutation of a stop cassette in MLH3. Mutations were inherited over many generations, demonstrating our ability to establish stable mutant lines. The availability of heritable CRISPR induced mutation in Brachionus rotifers now makes it possible to test mitochondrial mechanisms of maternal age effects more directly. The goal of this small-scale, short-term Administrative Supplement project is to establish 5-7 stable knockout mutants and knock-in fluorescent reporter strains for mitochondria-related genes in Brachionus manjavacas. Establishing mutants for genes related to mitochondrial function and dynamics will allow us to test the role of specific mitochondrial dynamics and metabolic pathways in maternal age effects on lifespan and healthspan in the parent R01 and future studies.