Functional Genomics of Mammalian Hibernation

项目总结 哺乳动物冬眠是一种综合的组织适应，包括协调协调 导致行为状态发生季节性变化的系统生理学。作为体温 沙鼠的常温值从36-38摄氏度下降到2摄氏度或更低，心率从200- 300次/分钟到7-10次/分钟，呼吸频率从100-150次/分钟下降到1-2次/分钟，以及 新陈代谢下降到常温值的1/30到1/100。这些机制构成了进入， 人们对冬眠的唤醒知之甚少。然而，分子、神经生物学和内分泌 这些戏剧性的唤醒状态变化背后的适应可能对人类健康具有重要意义 了解代谢动态平衡和衰老，以及加强器官保存的策略。 转录学和蛋白质组学研究已经开始描述这些基因背后的分子图景。 觉醒状态的变化。到目前为止，已发表的研究主要集中在一个器官系统上， 在跨系统集成方面几乎不费力气。在这个提案中，我们将利用12个被解剖的大脑 金毛黄鼠(Callopermophilus)各部位和10个外周组织的采集 在冬眠周期的5个主要阶段的12个时间点)创建一个公共数据库 来自相同个体的转录信息。在目标1中，基因组测序和从头组装 侧突圆线虫的基因组将由一名承包商进行，该承包商以前曾组装过13只- 线条地松鼠；合成的基因组将与国家实验室的同事合作进行注释 医学图书馆。这些努力将提供必要的脚手架，使目标2能够进行 对冬眠周期中采样的多个大脑区域进行RNAseq分析，并建立一个公共 转录信息数据库。我们的研究最初将集中在基底前脑、下丘脑、脑桥、 和大脑皮层，因为它们在唤醒状态调节中扮演着不同的角色。此RNAseq数据将丰富 在AIM 1中进行基因组注释。RNAseq数据将被可视化并可用于挖掘 通过一个公开可用的数据库，类似于CircaDB(http://circadb.hogeneschlab.org)，，基因表达 在昼夜节律生物学中广泛使用的数据库，我们将在Aim 2b中创建。数据库将是可扩展的， 支持添加来自其他大脑区域和外周器官的转录和蛋白质组数据 团体以及其他人。总而言之，这些信息将推动假说检验和整个生物体 冬眠期间发生的生理动态变化的分子基础的询问 循环，并可能提供对支持代谢抑制和阻止代谢抑制的适应的洞察 冬眠者可防止中风、呼吸暂停、肌肉萎缩和记忆力丧失。

PROJECT SUMMARY Mammalian hibernation is an integrative organismal adaptation involving a coordinated orchestration of systems physiology that results in a seasonally expressed change in behavioral state. As body temperature declines from euthermic values of 36-38°C to 2°C or lower in the ground squirrel, heart rate drops from 200- 300 beats/min to 7-10 beats/min, respiration rates fall from 100-150 breaths/min to 1-2 breaths/min, and metabolism declines to 1/30 to 1/100 of euthermic values. The mechanisms that underlie the entrance into, and arousal from, hibernation are poorly understood. However, the molecular, neurobiological, and endocrine adaptations underlying these dramatic arousal state changes are likely to have significance for human health in understanding metabolic homeostasis and aging, as well as for strategies to enhance organ preservation. Transcriptomic and proteomic studies have begun to describe the molecular landscape that underlies these changes in arousal state. To date, published studies have primarily focused on one organ system at a time, with little effort at integration across systems. In this proposal, we will exploit a bank of 12 dissected brain regions and 10 peripheral tissues collected from the golden-mantled ground squirrel (Callospermophilus lateralis) at 12 time points across the 5 major phases of the hibernation cycle to create a public database of transcriptomic information from the same individuals. In Aim 1, genome sequencing and de novo assembly of the C. lateralis genome will be conducted by a contractor who has previously assembled the genome of the 13- lined ground squirrel; the resultant genome will be annotated in collaboration with colleagues at the National Library of Medicine. These efforts will provide the necessary scaffold to enable Aim 2, whose goal is to conduct RNAseq analyses of multiple brain regions sampled across the hibernation cycle and to establish a public database of transcriptomic information. Our studies will initially focus on basal forebrain, hypothalamus, pons, and cerebral cortex because of their distinct roles in arousal state regulation. This RNAseq data will enrich the genome annotation conducted in Aim 1. The RNAseq data will be visualized and made available for mining through a publicly available database similar to CircaDB (http://circadb.hogeneschlab.org), a gene expression database widely used in circadian biology, that we will create in Aim 2b. The database will be extensible, enabling addition of transcriptomic and proteomic data from other brain regions and peripheral organs by our group as well as others. Collectively, this information will fuel hypothesis testing and organism-wide interrogation of the molecular bases of the dynamic changes in physiology that occur across the hibernation cycle and may provide insights into the adaptations that enable metabolic suppression and those that prevent hibernators from experiencing stroke, apnea, muscle atrophy and memory loss.