Neuronal Fatigue in Aging Hippocampus during Sustained Metabolic Demand

描述(由申请人提供)：衰老可能与对代谢挑战的反应能力降低有关，例如，导致认知任务的疲劳或对底物剥夺的敏感性增加，如相对低血糖或缺氧。我们假设，细胞和分子在调节固有的中枢神经系统生物能量学(即神经元-神经胶质相互作用、有氧和无氧代谢途径、底物可获得性等)中发生了变化。在衰老过程中会损害对持续能源需求的适应性反应。我们建议在没有系统性因素的情况下，研究老年动物分离的脑组织的长期代谢需求和疲劳，以确定神经元代谢和神经元-神经胶质相互作用的内在变化。在体外，从老年动物获得的脑片保留了该年龄在体内的代谢特征，以及导致局部组织规模新陈代谢调节的内在回路和其他因素。初步实验表明，与年轻动物的组织相比，老化海马区的神经元功能和线粒体氧化还原状态更容易受到代谢应激的影响，如葡萄糖水平降低和突触刺激时间延长，这表明老年人在较长一段时间内支持氧化新陈代谢速度加快的能力可能会降低。我们将通过直接测量组织乳酸、葡萄糖和PO2、NAD(P)H荧光和海马神经元反应来研究氧利用、线粒体氧化还原状态和神经元活动之间的能量关系，以评估长期代谢应激期间的神经元疲劳和神经元-神经胶质细胞相互作用。这些技术将在长时间的突触刺激(代谢需求增加)和有限的底物输送条件下使用。这些结果将有助于理解局部组织反应和生物能量学如何影响衰老过程中的新陈代谢。对神经元疲劳潜在机制的了解可能会为治疗提供新的靶点，从而可能提高持续认知任务的表现。 公共卫生相关性：这项建议试图了解大脑中的新陈代谢如何随着年龄的变化而变化，评估对持续反应的疲劳的潜在机制和可能的治疗方向。我们的目标是评估氧化和糖酵解代谢的成分，特别是在持续的代谢需求期间，超过几分钟，这是神经元和胶质细胞固有的。这里提出的体外切片制备方法可以直接评估脑组织中的局部代谢相互作用，而不直接涉及血管系统和系统提供底物。

DESCRIPTION (provided by applicant): Aging can be associated with a decreased ability to respond to metabolic challenges resulting, for example, in fatigue on cognitive tasks or increased susceptibility to substrate deprivation, such as relative hypoglycemia or hypoxia. We hypothesize that cellular and molecular changes in the regulation of intrinsic CNS bioenergetics (i.e., neuronal-glial interactions, aerobic and anaerobic metabolic pathways, substrate availability, etc.) during aging can impair adaptive responses to sustained energy demand. We propose to study prolonged metabolic demand and fatigue in isolated brain tissue from aged animals, in the absence of systemic factors such as poor vasculature or systemic substrate supply, to identify intrinsic changes in neuronal metabolism and neuronal-glial interactions. In vitro brain slices obtained from aged animals retain the in vivo metabolic characteristics of that age, as well as the intrinsic circuits and other factors leading to regulation of metabolism on a local tissue scale. Preliminary experiments indicate that neuronal function and mitochondrial redox state in aging hippocampus are more vulnerable to metabolic stress, such as lowered glucose levels and prolonged synaptic stimulation, compared to tissue from younger animals, suggesting that aged individuals may have reduced ability to support an increased rate of oxidative metabolism for an extended period of time. We will evaluate neuronal fatigue and neuronal-glial interactions during prolonged metabolic stress by studying the energetic relationships between oxygen utilization, mitochondrial redox state, and neuronal activity, using direct tissue lactate, glucose and Po2 measurements, NAD(P)H fluorescence, and neuronal responses in hippocampus. These techniques will be used during prolonged synaptic stimulation (increased metabolic demand) and conditions of limited substrate delivery. These results will facilitate understanding how local tissue responses and bioenergetics affect metabolism in aging. The understanding of the mechanisms underlying neuronal fatigue may indicate novel targets for treatment which may enhance performance on sustained cognitive tasks. PUBLIC HEALTH RELEVANCE: This proposal seeks to understand how metabolism in the brain changes with aging, assessing both mechanisms underlying fatigue to persistent responses and possible treatment directions. The goal is to assess components of oxidative and glycolytic metabolism, particularly during sustained metabolic demand, over minutes, which are intrinsic to neurons and glia. The in vitro slice preparation proposed here allows assessment of local metabolic interactions directly in brain tissue, without direct involvement of the vascular system and systemic provision of substrates.