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.

水环境不断受到温度变化、药品和农药的挑战。我们刚刚开始了解这些压力源如何在短期和长期内影响动物生理。迄今为止，大多数研究都集中在生命早期暴露于压力源或成年人的急性暴露。很少有人关注生命周期暴露及其对生理的影响，更少有人探讨这些影响是否会在多代人中传播。我的研究计划的长期愿景是了解生长，代谢和压力轴的综合反应，预测水生生物如何应对其一生中的复杂挑战。我将探讨挑战如何影响线粒体功能和能量利用，以及这些影响是否是多代或跨代的。我工作的第一个短期目标将集中在探索这些系统如何受到药物二甲双胍（Met）和农药氯硝柳胺（Nic）的影响，从受精到成年，因为两者都会损害线粒体功能和能量代谢。第二个目标将侧重于探索暴露是否具有多代效应，通过母系或父系，使用斑马鱼作为模式生物。为此，动物将从受精到成年阶段暴露于Met和Nic。将收集组织，并将制定固相萃取方案，利用液相色谱-飞行时间-质谱法测量鱼类组织中的化学物质。HQP将进行终点和动力学分析的组织处理，以测量组织代谢能力、能量储备，并将进行线粒体分离，以确定生命周期暴露如何影响不同发育阶段的功能。将在男性与女性效应的背景下探索这些终点，以研究反应的性别特异性差异。为了研究Nic和Met对动物从基因到整个生物体的影响，我们将在鱼类中进行实时定量PCR，ELISA，类固醇激素测量和耗氧量测定。接下来，通过与暴露和未暴露的亲本谱系进行配对杂交，将在后续世代中测量这些参数。该计划将为HQP提供综合，跨学科和实践经验，从事与环境相关的项目。HQP将获得分子和分析化学技能，将能够进行生化分析和细胞器分离技术，所有这些都与环境生理学，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终生压力的理解，并使我们能够更好地预测种群水平的影响。HQP将接受可转移技能的培训，如偏见意识，多种认知方式，科学中的性别偏见，并将被引入加拿大的环境研究。