Unraveling the mechanism of cryptic sexual cycle in Amoeba, Cochliopodium, using comparative transcriptome and immunocytochemistry

利用比较转录组和免疫细胞化学揭示阿米巴、Cochliopodium 神秘性周期的机制

• 批准号: 8957502
• 负责人: Yonas Tekle
• 金额: $ 33.58万
• 依托单位: SPELMAN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-18 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8957502
• 关键词: Address; Amoeba genus; Animal Model; Animals; Behavior; Bioinformatics; Biological Models; Biological Process; Cell Differentiation process; Cell physiology; Communicable Diseases; Complex; Custom; Cyst; Cytokinesis; Data; Development; Dictyostelium discoideum; Disease; Entamoeba histolytica; Environment; Eukaryota; Evolution; Exhibits; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genetic Recombination; Genetic Variation; Genome; Genomics; Health; Human; Knowledge; Life; Life Cycle Stages; Light; Mediating; Meiosis; Methods; Microbe; Molecular; Molecular Profiling; Nature; Nuclear Fusion; Partner in relationship; Pathogenesis; Pharmaceutical Preparations; Physarum polycephalum; Plants; Play; Ploidies; Reproduction; Role; Series; Sexual Development; Sexuality; Signal Transduction; Staging; System; Testing; Virulence; Work; asexual; base; cell motility; combat; comparative; fitness; genetic analysis; immunocytochemistry; insight; microbial; new therapeutic target; next generation sequencing; novel; pathogen; plant fungi; pressure; public health relevance; reproductive; research study; sex; tool

 DESCRIPTION (provided by applicant): Unraveling the Mechanism of Cryptic Sexual Cycle Development in Amoeba, Cochliopodium, using Comparative Transcriptome and Immunocytochemistry The proposed work aims to unravel the cytological and genetic underlying of the cryptic sexual life cycle of an amoeboid lineage, Cochliopodium. Sexual reproduction is highly beneficial in eukaryotes by enabling them to increase: genetic diversity, fitness under selective pressure and virulence as in pathogenic eukaryotes. However, sexuality in most microbial eukaryotes is poorly understood. This is mainly due to their diverse quality and complex life cycle. Unlike most other amoebae including E. histolytica that are assumed to engage in sex during the dormant, cyst stage (a challenge for experimental study), Cochliopodium undergoes nuclear fusion in the actively growing vegetative stage. This makes it an ideal candidate for investigative work involving live experimentation. The proposed work will test three interrelated hypotheses including genetic mechanisms of sexual development, mechanisms of genome reduction and genetic recombination and mating system in Cochliopodium. We will use a combination of methods including comparative transcriptome obtained using next generation sequencing, cytological evidence on targeted sex genes using immunocytochemistry (ICC), and a series of culturing experimentation to understand mating system in Cochliopodium. We have already built bioinformatics pipeline and custom scripts to assemble and analyze the genetic data. Newly available statistical tools for de novo data will be used for comparative gene expression analysis. Addressing the proposed hypotheses will enable: (a) uncover cytological signatures of the stages of sexual development in Cochliopodium, (b) shed light on the molecular mechanisms of a possible cryptic sexual life cycle in Cochliopodium and others, (c) understand mechanisms of genome reduction and genetic recombination in cryptic life cycles, (d) exploration of targets that might be used to combat eukaryotic pathogens, and (e) provide insights into the origin and evolution of sexuality in microbial eukaryotes.

 描述（由申请人提供）：使用比较转录组和免疫细胞化学揭示变形虫（Cochliopodium）中隐藏性周期发育的机制拟议的工作旨在揭示变形虫谱系（Cochliopodium）隐藏性生活周期的细胞学和遗传学基础。有性生殖在真核生物中是非常有益的，使它们能够增加：遗传多样性，选择压力下的适应性和致病性真核生物中的毒力。然而，大多数微生物真核生物的性行为知之甚少。这主要是由于其多样的质量和复杂的生命周期。与包括E.在被认为在休眠期、包囊期进行性行为的溶组织藻中，旋足藻在活跃生长的营养生长期进行核融合。这使其成为涉及现场实验的调查工作的理想候选人。本研究将对旋足藻有性发育的遗传机制、基因组减数分裂的遗传机制以及遗传重组和交配系统等三个相互关联的假说进行验证。我们将使用一系列的方法，包括比较转录组获得下一代测序，细胞学证据的目标性别基因的免疫细胞化学（ICC），和一系列的培养实验，以了解交配系统在旋足藻。我们已经建立了生物信息学管道和自定义脚本来组装和分析遗传数据。将使用新的从头数据统计工具进行比较基因表达分析。处理拟议的假设将能够：（a）揭示旋足藻属有性发育阶段的细胞学特征，（B）阐明旋足藻属和其它植物中可能的隐蔽性性生活周期的分子机制，（c）了解隐蔽性生活周期中基因组减少和遗传重组的机制，（d）探索可能用于对抗真核病原体的靶点，以及（e）提供对微生物真核生物中性的起源和进化的深入了解。