Molecular regulation of diapause in Culex pipiens

淡色库蚊滞育的分子调控

• 批准号: 7387334
• 负责人: DAVID L DENLINGER
• 金额: $ 20.86万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7387334
• 关键词: Adult; Birds; Blood; Categories; Cues; Culex pipiens; Culicidae; Decision Making; Development; Diapause; Facility Construction Funding Category; Fatty Acids; Female; Gene Expression; Genes; Housing; In Situ Hybridization; Juvenile Hormones; Length; Link; Malate Dehydrogenase; Methoprene; Molecular; Northern Blotting; Pathway interactions; Pattern; Phase; Physiological; Physiology; Polymerase Chain Reaction; Population; RNA Interference; Regulation; Resistance; Ribosomal Proteins; Seasonal Variations; Site; Staging; Techniques; Time; Tissues; Trypsin; Viral; Virus; West Nile virus; Work; base; beta Tubulin; chymotrypsin; design; feeding; gene function; novel; pathogen; programs; reproductive; research study; sugar; vector

DESCRIPTION (provided by applicant): This proposal focuses on the molecular basis for diapause (dormancy) in the northern house mosquito, Culex pipiens. In the early autumn adult females of this species seek protected sites and remain there throughout the winter months in a reproductively inactive state know as diapause. Although many aspects of the physiology and environmental control of diapause are well known, there is little information concerning the molecular basis for this diapause or the diapause of any other vector species. Obtaining such information is critical not only for understanding regulation of seasonal cycles of mosquitoes, but also for understanding the seasonality of pathogens the mosquito transmit. In the case of Cx. pipiens the overwintering females appear to harbor West Nile virus, and thus serve as the mechanism for reintroducing the virus into bird populations the following spring. Our initial studies using suppressive subtractive hybridization (SSH) have identified four categories of genes that are upregulated in early diapause. These include genes involved in the diapause-associated switch from blood feeding to sugar feeding, the arrest of development, cold-hardening, and responding to juvenile hormone. Similar SSH experiments will examine mid and late stages of diapause. Clones isolated with this technique will be further characterized by full-length sequencing, by evaluating expression patterns throughout diapause using northern blotting and real-time PCR, by examining tissue-specific expression patterns, and by evaluating gene function using RNA interference. We also plan to use SSH to search for genes associated with the programming phase of diapause (early pupal stage). Such genes are likely to be involved in the decision to either enter or avert diapause. At the conclusion of this study we expect to have identified major gene pathways involved in the regulation of diapause in Cx. pipiens, thus allowing us to begin the construction of a regulatory hierarchy for diapause in this species. We anticipate that these results will suggest vulnerable targets for disrupting diapause and may provide clues for how viral replication is halted during the winter months.

描述（由申请人提供）：本提案侧重于北方家蚊（尖音库蚊）滞育（休眠）的分子基础。在初秋，这个物种的成年雌性会寻找保护地，并在整个冬季停留在那里，处于一种被称为滞育的繁殖不活跃状态。虽然滞育的生理学和环境控制的许多方面是众所周知的，但关于这种滞育或任何其他媒介物种滞育的分子基础的信息很少。获得这些信息不仅对于了解蚊子季节性周期的调节至关重要，而且对于了解蚊子传播的病原体的季节性也至关重要。在CX的情况下。越冬的雌蚊似乎携带西尼罗河病毒，因此成为次年春天将病毒重新引入鸟类种群的机制。 我们使用抑制性消减杂交（SSH）的初步研究已经确定了四类基因在滞育早期上调。这些基因包括与滞育相关的从血液喂养到糖喂养的转换，发育停滞，冷硬化和对保幼激素的反应。类似的SSH实验将检查滞育的中期和晚期阶段。用这种技术分离的克隆将通过全长测序、通过使用北方印迹和实时PCR评估整个滞育期间的表达模式、通过检查组织特异性表达模式以及通过使用RNA干扰评估基因功能来进一步表征。我们还计划使用SSH来搜索与滞育（早期蛹阶段）的编程阶段相关的基因。这些基因可能参与决定进入或避免滞育。 在这项研究的结论，我们希望已经确定了主要的基因途径参与调控滞育的Cx。pipiens，从而使我们能够开始建设一个监管层次滞育在这个物种。我们预计，这些结果将提示破坏滞育的脆弱目标，并可能为病毒复制如何在冬季停止提供线索。