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Vibrio fischeri as a Model for Bacterial Colonization

费氏弧菌作为细菌定植的模型

基本信息

批准号：
10338077
负责人：
MARGARET J MC FALL-NGAI
金额：
--
依托单位：
UNIVERSITY OF HAWAII AT MANOA
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-12-01 至 2022-02-02
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10338077
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

项目摘要

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仅需 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开始和持续； (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。