Mechanisms of coping with a lifetime of environmental challenges: Integrating growth, metabolism and stress physiology across multiple generations

应对一生环境挑战的机制：整合多代人的生长、新陈代谢和应激生理学

• 批准号: RGPIN-2022-04109
• 负责人: Birceanu, Oana
• 金额: $ 2.11万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=763335
• 关键词: Mechanisms; coping; lifetime; environmental; challenges

Aquatic environments are constantly challenged by temperature shifts, pharmaceuticals and pesticides. We are just starting to understand how these stressors impact animal physiology over short and long-term. Most studies to date focus on either early life stage exposure to the stressors or on acute exposure of adults. Few focus on lifecycle exposure and how it impacts physiology and even fewer explore whether the effects are transmitted across multiple generations. The long-term vision of my research program is to understand how integrated responses at the growth, metabolism and stress axes, predict how aquatic organisms respond to complex challenges over their lifetime. I will explore how challenges impact mitochondria functioning and energy utilization, and whether these effects are multi- or transgenerational. A first short-term goals of my work will focus on exploring how these systems are impacted by the pharmaceutical metformin (Met) and the pesticide niclosamide (Nic) following a lifetime of exposure, from fertilization to adulthood, since both impair mitochondrial function and energy metabolism. A second goal will focus on exploring whether the exposures have multigenerational effects, via the maternal or paternal lines, using zebrafish as model organisms. To this end, animals will be exposed to Met and Nic from fertilization to adult stages. Tissues will be collected and solid-phase extraction protocols will be developed to measure the chemicals in fish tissues using liquid chromatography-time-of-flight-mass spectrometry. HQP will conduct tissue processing for endpoint and kinetic assays to measure tissue metabolic capacity, energy reserves and will perform mitochondria isolations to determine how life-cycle exposure impacts function at various developmental stages. These endpoints will be explored in the context of male vs female effects, to investigate sex-specific differences in responses. To investigate the effects of Nic and Met on the animals, from genes to whole organism, we will perform real-time quantitative PCR, ELISAs, steroid hormone measurements and oxygen consumption assays in fish. Next, these parameters will be measured in subsequent generations, by doing paired crosses with exposed and non-exposed parental lineages. This program will provide HQP with an integrative, interdisciplinary and hands-on experience, working on an environmentally relevant project. HQP will gain molecular and analytical chemistry skills, will be able to conduct biochemical assays and organelle isolation techniques, all of which are relevant to the fields of environmental physiology, pharmacology and toxicology. This work will improve our understanding of how fish respond to a lifetime of stress and will allow us to better predict population-level effects. HQP will be trained in transferable skills, such as bias awareness, multiple ways of knowing, gender bias in sciences and will be introduced to environmental research in Canada.

水生环境不断受到温度转移，药物和农药的挑战。我们刚刚开始了解这些压力源在短期和长期内如何影响动物生理。迄今为止，大多数研究都集中在早期生命阶段暴露于压力源或成年人的急性暴露上。很少有人关注生命周期暴露及其如何影响生理学，甚至更少探索效果是否在多代人中传播。我的研究计划的长期愿景是了解如何在生长，代谢和压力轴上的综合反应预测水生生物在其一生中如何应对复杂挑战。我将探讨挑战如何影响线粒体的功能和能量利用，以及这些影响是多或转世的。我工作的第一个短期目标将集中于探索这些系统如何受到一生暴露于施肥到成年的终生暴露后的药物二甲双胍（MET）和农药烟酰胺（NIC）的影响，因为这既损害了线粒体功能和能量代谢。第二个目标将集中在探索使用斑马鱼作为模型生物的曝光是否具有通过母体或父亲线的多代效应。为此，动物将暴露于Met和NIC，从受精到成人阶段。将收集组织，并将使用液态色谱时光质谱法制定固相提取方案，以测量鱼类组织中的化学物质。 HQP将进行组织加工以进行端点和动力学测定，以测量组织代谢能力，能量储量并进行线粒体分离，以确定生命周期暴露在各个发育阶段的影响如何。这些终点将在男性与女性效应的背景下进行探讨，以研究针对性的性别差异。为了研究NIC和MET对动物的影响，从基因到整个生物体，我们将进行实时定量PCR，ELISA，类固醇激素测量和鱼类中的氧气消耗测定法。接下来，将通过与暴露和不暴露的父母谱系进行配对的十字架来测量这些参数。该计划将为HQP提供综合，跨学科和动手的经验，从事与环境相关的项目的工作。 HQP将获得分子和分析化学技能，能够进行生化测定和细胞器隔离技术，所有这些技术与环境生理学，药理学和毒理学领域有关。这项工作将提高我们对鱼类如何应对一生的压力的理解，并使我们能够更好地预测人口水平的影响。 HQP将接受可转移技能的培训，例如偏见意识，多种认识方式，科学的性别偏见，并将被引入加拿大的环境研究。