Vibrio fischeri as a Model for Bacterial Colonization

费氏弧菌作为细菌定植的模型

• 批准号: 10089379
• 负责人: MARGARET J MC FALL-NGAI
• 金额: $ 51.27万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10089379
• 关键词: Address; Affect; Animals; Apical; Automobile Driving; Bacteria; Bacterial Model; Binding; Biochemical; Biological Models; Biology; Biophysics; Birth; Cell Nucleus; Cells; Cellular biology; Characteristics; Chronic; Circadian Rhythms; Communities; Complex; Confocal Microscopy; Cues; DNA; Development; Environment; Epithelial; Euprymna scolopes; Event; Exposure to; Fluorescent in Situ Hybridization; Fostering; Gastrointestinal tract structure; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Genomics; Goals; Gram-Negative Bacteria; Health; Hour; Human; Human Microbiome; Image; Immune response; Individual; Investigation; Libraries; Life; Light; Maintenance; Measures; Membrane; Methods; Microbe; Microbial Genetics; Modeling; Molecular; Mucous Membrane; Nanotechnology; Nuclear; Outcome; Pathogenesis; Pathogenicity; Pattern; Process; RNA; Reaction; Research; Resolution; Role; Scientist; Series; Signal Transduction; Signaling Molecule; Small RNA; Spectrometry, Mass, Secondary Ion; Squid; Study models; Surface; Symbiosis; System; Time; Tissues; Transcript; V. fischeri-squid system; Variant; Vesicle; Vibrio; Vibrio cholerae; Vibrio fischeri; Virulence; Visualization; beneficial microorganism; critical period; extracellular; frontier; gastrointestinal epithelium; host colonization; host microbiome; host-microbe interactions; human tissue; infancy; innovation; insight; microbial; microbial community; microbial composition; microbiota; nano-string; nanoscale; new technology; normal microbiota; novel imaging technique; novel strategies; pathogen; pathogenic microbe; programs; response; success; symbiont; trafficking; transcriptome sequencing; transcriptomics

Project Summary/Abstract - The long-term goals of the proposed research program are to provide insight into the complex dialogue that occurs each generation between humans and their microbiota during the critical period following birth. Recent research has indicated a strong correlation between outcomes of these early events and life-long health. However, the inaccessibility of colonized tissues and high diversity of the microbiota renders an in-depth study of early colonization of human tissues extremely challenging. When faced with such complex phenomena, biologists often turn to simpler model systems to provide insights into evolutionarily conserved features and reveal basic principles. To decipher the cellular and molecular mechanisms underlying the initiation of bacterial associations with apical surfaces of mucosal epithelia, the proposed program exploits the binary symbiosis between the bacterium Vibrio fischeri and its squid host, Euprymna scolopes. This relatively simple association has been studied for over two decades as a model for the chronic colonization of mucosa by Gram-negative bacteria. As in humans, the squid-vibrio association begins anew each generation, requiring a `winnowing' of other environmental bacteria that results in persistent association restricted to the coevolved partners. In this system, the process of symbiosis initiation occurs across ~100 microns over minutes to hours. It can be directly imaged in its entirety using confocal microscopy, which offers the rare opportunity to define, with high temporal and spatial resolution, the reciprocal molecular and biochemical dialogue that results in the establishment of a specific, life-long beneficial symbiosis. This project brings together two collaborators, each with expertise in the biology of one of the symbiotic partners, and introduces new technology to the study of host-microbe interactions, including: Nanoscale Secondary Ion Mass Spectrometry (NanoSIMS), which allows precision tracking of symbiont molecules into host tissues; Hybridization-Chain-Reaction Fluorescent In Situ Hybridization (HCR-FISH), which enables visualization of rare transcripts in host and symbiont cells; NanoString, a new technology for simultaneous analysis of dozens to hundreds of targeted transcripts; and high-efficiency RNAseq, which produces robust transcriptional libraries from as little as 10 ng total RNA (~105 bacteria). Specific aims to be addressed are: (1) the examination of how symbiotic bacterial strain variation affects symbiosis onset and persistence; (2) the characterization of outer membrane vesicle (OMV) contents, their trafficking into host cells, and host responses to OMV cargo; and, (3) the investigation of the roles of vibrio virulence determinants in a non-pathogenic association. An understanding of the human microbiome is in its infancy, and this frontier field is currently at the stage of building paradigms. Within this context, as the squid-vibrio system has in the past, the results of the current study will shed light upon fundamental principles governing the onset of both beneficial and pathogenic associations.

项目摘要/摘要 - 拟议研究计划的长期目标是深入了解 在关键时期，每一代人与其微生物群之间发生的复杂对话 出生后的时期。最近的研究表明，这些早期的结果之间存在很强的相关性。 事件和终身健康。然而，定植组织的不可接近性和高度多样性 微生物群使得深入研究人体组织的早期定植变得极具挑战性。当面对 对于如此复杂的现象，生物学家经常转向更简单的模型系统来提供见解 进化上保守的特征并揭示基本原理。破译细胞和分子 细菌与粘膜上皮顶端表面关联的启动机制 拟议的程序利用了费氏弧菌及其鱿鱼宿主之间的二元共生关系， Euprymna scolopes。这种相对简单的关联作为模型已经被研究了二十多年。 革兰氏阴性菌在粘膜上的慢性定植。与人类一样，鱿鱼-弧菌协会 每一代都重新开始，需要对其他环境细菌进行“筛选”，从而导致持久的 关联仅限于共同进化的伙伴。在这个系统中，发生共生启动过程 在几分钟到几小时内跨越约 100 微米。它可以使用共焦显微镜直接整体成像， 这提供了难得的机会，以高时间和空间分辨率定义倒数分子 和生化对话，从而建立特定的、终生有益的共生关系。这 该项目汇集了两名合作者，每人都拥有共生伙伴之一的生物学专业知识， 并引入新技术来研究宿主-微生物相互作用，包括： 纳米级二次离子 质谱（NanoSIMS），可以精确追踪宿主组织中的共生分子； 杂交链反应荧光原位杂交 (HCR-FISH)，可实现可视化 宿主和共生细胞中的稀有转录本； NanoString，一种可同时分析数十种物质的新技术 数百个目标转录本；和高效 RNAseq，可产生强大的转录文库 来自低至 10 ng 总 RNA（约 105 个细菌）。需要解决的具体目标是：（1）检查如何 共生菌株变异影响共生的发生和持续； (2)外在表征 膜囊泡 (OMV) 内容物、它们进入宿主细胞的运输以及宿主对 OMV 货物的反应；并且，(3) 研究弧菌毒力决定因素在非致病性关联中的作用。一个 对人类微生物组的了解还处于起步阶段，这一前沿领域目前正处于 构建范式。在此背景下，正如过去的鱿鱼弧菌系统一样，当前的结果 研究将揭示控制有益和致病的发生的基本原则 协会。