Functional Genomics of Mammalian Hibernation

项目摘要 哺乳动物的冬眠是一种综合性的生物适应，涉及到协调的 导致行为状态季节性变化的系统生理学。体温 从36-38°C的适温值下降到2°C或更低，心率从200- 200 ° C下降到200- 200 ° C。 300次/分降至7-10次/分，呼吸频率从100-150次/分降至1-2次/分，以及 代谢下降至正常体温值的1/30至1/100。入口的机制， 以及从冬眠中苏醒的机制还不太清楚然而，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， 了解代谢稳态和衰老，以及提高器官保存的策略。 转录组学和蛋白质组学研究已经开始描述这些基础的分子景观 唤醒状态的变化迄今为止，已发表的研究主要集中在一个器官系统的时间， 在跨系统集成方面几乎没有努力。在这个提议中，我们将利用12个解剖的大脑库 从金背地松鼠（Callospermophilus）的10个区域和10个外周组织中收集 在冬眠周期的5个主要阶段的12个时间点， 来自同一个体的转录组信息。在目标1中，基因组测序和从头组装 梭lateralis基因组将由一个承包商进行，该承包商先前已组装了13- 线地松鼠;由此产生的基因组将与国家科学院的同事合作注释 医学图书馆。这些努力将为实现目标2提供必要的框架，目标2的目标是进行 RNAseq分析了在冬眠周期中采样的多个大脑区域，并建立了一个公共的 转录组信息数据库。我们的研究将首先集中在基底前脑，下丘脑，脑桥， 和大脑皮层，因为它们在唤醒状态调节中的独特作用。该RNAseq数据将丰富 Aim 1中进行的基因组注释。RNAseq数据将被可视化并可用于挖掘 通过类似于CircaDB（http：//www.example.com）的公开可用的数据库，circadb.hogeneschlab.org 我们将在目标2b中创建一个广泛应用于昼夜节律生物学的数据库。数据库将是可扩展的， 使我们能够添加来自其他大脑区域和外周器官的转录组和蛋白质组数据， 集团以及其他。总的来说，这些信息将推动假设检验和整个生物体的研究。 询问冬眠期间生理学动态变化的分子基础 循环，并可能提供洞察适应，使代谢抑制和那些防止 冬眠者经历中风，呼吸暂停，肌肉萎缩和记忆丧失。

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.