Whole-lifespan monitoring of yeast health

到2060年，65岁及以上的美国人比例预计将翻一番。 治疗与年龄有关的疾病，这一根本的人口变化将预示着激烈的经济和 社会挑战。对S.酿酒厂揭示了最常见的 死亡原因包括癌症、心血管疾病、神经变性和糖尿病。 使用微流控装置，我们可以通过延时以单细胞分辨率监测酵母老化过程 显微镜该设备还允许我们研究老化细胞的环境扰动。微流体捕集 是一种罕见的有效方法，用于收集单电池的纵向，动态，原位老化数据 分辨率我将利用这个设备进行酵母细胞的全寿命监测， 具体目标： 目的1：探讨年龄相关性线粒体功能障碍中铁饥饿反应的机制 线粒体功能障碍被认为是多种与年龄相关的疾病的驱动因素，17并且 线粒体表型与酵母RLS有机械联系。 19初步数据表明，铁离子在线粒体中的作用， 饥饿反应介导了这一过程。我将肯定液泡酸度，铁调节， 线粒体功能障碍，并确定这些连接的因果机制。 目的2：确定年龄相关的母亲-女儿寿命不对称损失的机制 不对称的细胞分裂是后生动物生命的标志，在发育和 与酵母衰老领域的流行假设相反，我观察到一种与酵母衰老相关的病理学。 随着年龄的增长，母亲与女儿的寿命不对称性逐渐下降，这种不对称性在生命早期就开始了。我 假设这种下降是由于septin介导的皮质和膜的进行性下降， 扩散阻挡层我将研究这种不对称性丧失的动力学和机制。 目的3：确定年龄相关的保真度损失在MSN 2葡萄糖传感信号转导中的作用 衰老的特征是对环境变化的反应能力降低，维持体内平衡的能力下降。 我们的微流控装置使扰动和观察衰老细胞成为可能。我发现MSN 2信号 在相同的环境中，年轻细胞和老年细胞之间存在差异。此外，我发现暴露于 随着年龄的增长，随时间变化的环境具有较短的寿命。我假设MSN 2输了 葡萄糖信号传导保真度，因为它在报告随着年龄的增加氧化应激中的作用。我会 描述这种信息传递的损失，并确定它是否是负责与年龄有关的损失， resilience.

The fraction of Americans 65 and older is expected to double by 2060.13 Unless significant progress is made in the treatment of age-related diseases, this fundamental demographic shift will herald drastic economic and social challenges. The study of aging in S. cerevisiae has revealed fundamental insights into the most common causes of mortality including cancer, cardiovascular disease, neurodegeneration, and diabetes.14–16 Using a microfluidic device, we can monitor the yeast aging process at single-cell resolution via time-lapse microscopy. This device also allows us to study environmental perturbations of aged cells. Microfluidic trapping is a rare validated method for collection of longitudinal, dynamic, in situ aging data at single-cell resolution. I will exploit this device to perform whole-lifespan monitoring of yeast cell towards the following specific aims: Aim 1: Determine mechanism of iron starvation response in age-related mitochondrial dysfunction Mitochondrial dysfunction has been implicated as a driver of multiple age-related diseases,17 and changes in mitochondrial phenotype have been mechanistically tied to yeast RLS.18 Previous research has linked an early life loss of vacuolar acidity to later mitochondrial dysfunction.19 Preliminary data indicates that an iron starvation response mediates this process. I will affirm the links between vacuolar acidity, iron regulation, and mitochondrial dysfunction and determine the causal mechanism of these connections. Aim 2: Determine mechanism of age-related loss of mother-daughter lifespan asymmetry Asymmetric cell division is a hallmark of metazoan life, with important implications in both development and age-related pathology.20,21 Contrary to popular assumption in the yeast aging field, I have observed a progressive age-dependent decline in mother-daughter lifespan asymmetry that begins early in life. I hypothesize that this decline is due to a progressive decline in the septin-mediated cortical and membrane diffusion barrier. I will investigate the kinetics and the mechanism of this loss of asymmetry. Aim 3: Define role of age-related loss of fidelity in MSN2 glucose-sensing signal transduction Aging is characterized by a reduced ability to maintain homeostasis in response to environmental change.22,23 Our microfluidic device makes it possible to perturb and observe aged cells. I have found that MSN2 signaling differs between young and old cells in the same environment. Moreover, I have found that cells exposed to temporally varying environments as they age have shorter lifespans. I hypothesize that the MSN2 loses glucose-signaling transmission fidelity due to its role in reporting of increased oxidative stress with age. I will characterize this loss of information transfer and determine whether it is responsible for an age-related loss of resilience.