Longevity and Stress Resistance

描述(由申请人提供)：在父计划项目中，我们假设AC5中断(基因敲除，KO)是长寿的模型，并计划将其与研究最广泛的长寿模型卡路里限制(CR)进行比较。我们假设AC5KO和CR的结合可能会导致更长的寿命，如果涉及到足够不同的机制，可以协同作用。在该计划项目的头两年，对这一假设的检验得出了截然不同的结果，即叠加CR的AC5KO小鼠在一个月内全部死亡。此次修订的目的是深入研究调节这种致命结合的机制，因为共同的分子和细胞机制可能是调节长寿和抗逆性的关键分子机制。由于这两种模型都表现出显著的代谢变化，首先我们将研究响应有限的热量摄入或增强的代谢的主要机制，例如，葡萄糖利用、糖原储备的耗尽和脂肪酸的利用。接下来，我们将通过基因组和蛋白质组研究来检验这些主要的长寿和抗逆性模型所共有的分子机制。为了实现这一点，我们正在为该计划项目提出第四个项目。新项目(‘限制卡路里和AC5基因敲除中的共同长寿机制’)也将利用所有核心(核心A：管理/生理学、核心B：动物护理、核心C：蛋白质组/基因组学、核心D：生物信息学/生物统计学和核心E：病理学)。这一修订应用的中心假设是AC5抑制和CR通过相似的机制诱导长寿和应激抵抗，本项目将确定共同的和独特的共同机制，调节长寿和应激抵抗。 具体目的1：检测AC5KO小鼠血液、肝脏、心脏、骨骼肌和脂肪组织中葡萄糖和脂肪酸的代谢，并与中度和重度热量限制的WT进行比较。第二个目标将是比较AC5KO中两个水平的CR叠加的效果。 具体目的2：通过病理学、组织学和生化方法确定重叠CR的AC5KO小鼠的死亡原因。 具体目标3：利用基因组和蛋白质组学方法，确定AC5KO和CR共同调节寿命、抗逆性和新陈代谢的分子机制。

DESCRIPTION (provided by applicant): In the parent program project we had hypothesized that AC5 disruption (knock out, KO) was a model of longevity and planned to compare it to the most widely studied model of longevity, caloric restriction (CR). We hypothesized that the combination of AC5 KO and CR might result in even greater longevity, if there were sufficiently different mechanisms involved that could act synergistically. In the first two years of the program project the testing of this hypothesis resulted in dramatically different results, i.e., the AC5 KO mice with superimposed CR all died within one month. The goal of this revised application is to investigate the mechanisms in depth mediating this lethal combination, since the shared molecular and cellular mechanisms are likely the key molecular mechanisms mediating longevity and stress resistance. Since both models demonstrate significant alterations in metabolism, first we will examine the major mechanisms involved in response to limited caloric intake or to enhanced metabolism, e.g., glucose utilization, depletion of glycogen stores, and utilization of fatty acids. Next we will examine the molecular mechanisms shared by these major models of longevity and stress resistance by genomic and proteomic investigation. To accomplish this we are proposing a fourth Project for this Program Project. The new project ('Common Longevity Mechanisms in Caloric Restriction and AC5 Knockout') will also utilize all the Cores (Core A: Administration/ Physiology, Core B: Animal Care, Core C: Proteomics/Genomics, Core D: Bioinformatics/Biostatistics and Core E: Pathology). The central hypothesis of this revised revision application is that AC5 inhibition and CR induce longevity and stress resistance through similar mechanisms, and that this project will identify the shared mechanisms, which are both common and unique, mediating longevity and stress resistance. Specific Aim 1: To examine glucose and fatty acid metabolism in blood, liver, heart, skeletal muscle and adipose tissue in AC5 KO mice and compare to WT with moderate and severe caloric restriction. A second goal will be to compare the effects of superimposition of the two levels of CR in AC5 KO. Specific Aim 2: To determine the cause of death in AC5 KO mice with superimposition of CR using pathology, histology and biochemistry. Specific Aim 3: To determine shared molecular mechanisms that regulate longevity, stress resistance and metabolism in AC5 KO and CR using genomic and proteomic approaches.