Diapause in Culex Mosquitoes

库蚊滞育

• 批准号: 8076324
• 负责人: DAVID W SEVERSON
• 金额: $ 59.84万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8076324
• 关键词: Address; Adult; Arboviruses; Area; Behavior; Biochemical Pathway; Biological Assay; Biology; Candidate Disease Gene; Climate; Complex; Culex (Genus); Culex pipiens; Culicidae; Data Analyses; Developed Countries; Developing Countries; Development; Diapause; Disease; Disease Outbreaks; Drug resistance; Engineering; Epidemic; Experimental Designs; Female; Filaria bancrofti; Filarial Elephantiases; Foundations; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Crosses; Genetic Markers; Genetic Transcription; Genome; Genome Mappings; Genomics; Geographic Distribution; Goals; Health; Human; Individual; Insecticide Resistance; Knowledge; Laboratories; Lymphatic; Mediating; Microarray Analysis; Microsatellite Repeats; Molecular; Molecular Genetics; Molecular Profiling; Mosquito Control; Mosquito-borne infectious disease; Nematoda; North America; Parasites; Pattern; Phenotype; Population; Population Replacements; Procedures; Production; Protocols documentation; Quantitative Trait Loci; RNA Interference; Recombinants; Relative (related person); Research; Restriction fragment length polymorphism; Single Nucleotide Polymorphism; Single-Stranded Conformational Polymorphism; Surveys; Techniques; Time; Transgenes; Travel; Vaccines; Validation; Virus; West Nile virus; base; disease transmission; disorder control; disorder prevention; genome sequencing; genome-wide; novel; overexpression; pathogen; positional cloning; programs; research study; trait; transmission process; vaccine development; vector

DESCRIPTION (provided by applicant): Mosquito-borne diseases represent major health problems in most developing countries and have emerged as significant threats for epidemic outbreaks in developed countries. Exacerbating these problems are the emergence of insecticide resistance in mosquito populations, drug resistance among parasite populations, delayed progress in vaccine development, lack of support for mosquito control programs, and increased global travel. Mosquitoes in the Culex pipiens complex are primary vectors for the lymphatic filarioid nematode parasite, Wuchereria bancrofti and several arboviruses. This includes the recently introduced West Nile virus in North America. The long- term objectives of the proposed research are to identify, isolate and characterize genetic factors associated with diapause, a trait that strongly impacts the geographic distribution and survival of species within the Culex pipiens complex. At present, no knowledge exists on the molecular determinants of diapause in Culex. The objectives of this application are based on the general hypothesis that genes associated with the diapause phenotype can be identified and isolated by positional cloning techniques. Identification of genes determining diapause and the knowledge from the detailed understanding of the associated biochemical pathways could have several potential applications including: transgene constructs that promote altered and disease limiting behaviors following population replacement, development of targeted biopesticides, or identification of biologically relevant antagonists that disrupt behaviors critical to pathogen transmission or survival. The specific aims of the project are to: 1) fine-scale map genome regions identified as containing genes that determine the diapause phenotype, 2) identify candidate genes determining diapause, and 3) conduct functional analysis of candidate genes determining diapause. Information from genetic crosses will be leveraged against the Cx. quinquefasciatus genome project to identify candidate genes within discrete genome regions. Candidate genes will be evaluated using microarrays, followed by quantitative real time PCR assays to confirm and expand microarray results for promising genes. Positive candidate genes will be assayed for biological relevance using RNA interference mediated gene knockdowns or gene overexpression using recombinant Sindbis transducing viruses. Results from laboratory experiments will then be evaluated among natural populations representing Cx. pipiens s.s. and Cx. quinquefasciatus. PUBLIC HEALTH RELEVANCE: Mosquitoes in the Culex pipiens complex are primary vectors for human lymphatic filariasis and several arboviruses, including the West Nile virus. No vaccines are available and disease prevention is largely dependent on avoiding contact with infected mosquitoes. The proposed research will provide valuable new information on the biology of the mosquito relating to the ability of adult females to survive temperate climate winter in a diapause state.

描述（由申请人提供）：蚊媒疾病是大多数发展中国家的主要健康问题，并已成为发达国家流行病爆发的重大威胁。蚊子种群中杀虫剂耐药性的出现、寄生虫种群中耐药性的出现、疫苗开发进度的延迟、蚊子控制计划缺乏支持以及全球旅行的增加，加剧了这些问题。淡色库蚊复合体中的蚊子是淋巴丝状线虫寄生虫、班氏乌策菌和几种虫媒病毒的主要载体。这包括最近在北美引入的西尼罗河病毒。拟议研究的长期目标是识别、分离和表征与滞育相关的遗传因素，滞育是一种强烈影响淡色库蚊群内物种地理分布和生存的特征。目前，尚不清楚库蚊滞育的分子决定因素。本申请的目的基于这样的一般假设：可以通过定位克隆技术来鉴定和分离与滞育表型相关的基因。确定滞育基因的鉴定以及对相关生化途径的详细了解所获得的知识可能具有多种潜在应用，包括：在种群更替后促进改变和疾病限制行为的转基因构建体、靶向生物农药的开发或鉴定破坏的生物学相关拮抗剂对病原体传播或生存至关重要的行为。该项目的具体目标是：1）精细绘制基因组区域图谱，确定包含决定滞育表型的基因，2）识别决定滞育表型的候选基因，3）对决定滞育的候选基因进行功能分析。来自遗传杂交的信息将被用来对抗 Cx。 quinquefasciatus 基因组计划，用于识别离散基因组区域内的候选基因。将使用微阵列评估候选基因，然后进行定量实时 PCR 测定，以确认和扩展有希望基因的微阵列结果。将使用 RNA 干扰介导的基因敲低或使用重组 Sindbis 转导病毒的基因过度表达来分析阳性候选基因的生物学相关性。然后，实验室实验的结果将在代表 Cx 的自然群体中进行评估。皮皮安斯公司和Cx。致倦五线虫。公共卫生相关性：淡色库蚊复合体中的蚊子是人类淋巴丝虫病和多种虫媒病毒（包括西尼罗河病毒）的主要传播媒介。目前尚无疫苗可用，疾病预防在很大程度上取决于避免接触受感染的蚊子。拟议的研究将为蚊子的生物学提供有价值的新信息，这些信息与成年雌性在滞育状态下度过温带冬季的能力有关。