Molecular regulation of diapause in Culex pipiens

淡色库蚊滞育的分子调控

• 批准号: 7585282
• 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 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7585282
• 关键词: Adult; Birds; Blood; Categories; Cues; Culex pipiens; Culicidae; Decision Making; Development; Diapause; 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; 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 discovery; gene function; meetings; 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(申请人提供)：这项建议集中在北方家蚊滞育(休眠)的分子基础上，库蚊。在初秋，这一物种的成年雌性寻找受保护的地方，并在整个冬季的几个月里停留在那里，处于繁殖不活跃的状态，称为滞育。虽然滞育的生理和环境控制的许多方面是众所周知的，但关于这种滞育的分子基础或任何其他媒介物种的滞育的信息很少。获得这些信息不仅对于了解蚊子季节性周期的规律至关重要，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。越冬的雌性蚊子似乎携带西尼罗河病毒，因此成为第二年春天将病毒重新引入鸟类种群的机制。 我们利用抑制性消减杂交(SSH)进行的初步研究已经确定了四类在滞育早期上调的基因。这些基因包括与滞育相关的基因，包括从血液摄食到糖分摄食的转换，发育停滞，冷变硬，以及对保幼激素的反应。类似的SSH实验将检测滞育的中后期。用这项技术分离的克隆将通过全长测序、使用Northern blotting和实时PCR评估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。我们还计划利用SSH来寻找与滞育编程阶段(早期蛹阶段)相关的基因。这些基因很可能参与决定是否进入或避免滞育。 在这项研究的结论中，我们预计已经确定了参与调节Cx滞育的主要基因途径。从而允许我们开始构建该物种滞育的调节等级。我们预计，这些结果将提出破坏滞育的脆弱目标，并可能为病毒如何在冬季几个月停止复制提供线索。

项目成果

Catalase and superoxide dismutase-2 enhance survival and protect ovaries during overwintering diapause in the mosquito Culex pipiens.

• DOI: 10.1016/j.jinsphys.2011.01.012
• 发表时间: 2011-05
• 期刊: JOURNAL OF INSECT PHYSIOLOGY
• 影响因子: 2.2
• 作者: Sim, Cheolho;Denlinger, David L.
• 通讯作者: Denlinger, David L.

Meeting the challenges of on-host and off-host water balance in blood-feeding arthropods.

• DOI: 10.1016/j.jinsphys.2010.02.014
• 发表时间: 2010-10
• 期刊: JOURNAL OF INSECT PHYSIOLOGY
• 影响因子: 2.2
• 作者: Benoit, Joshua B.;Denlinger, David L.
• 通讯作者: Denlinger, David L.

A shut-down in expression of an insulin-like peptide, ILP-1, halts ovarian maturation during the overwintering diapause of the mosquito Culex pipiens.

• DOI: 10.1111/j.1365-2583.2009.00872.x
• 发表时间: 2009-06
• 期刊: Insect molecular biology
• 影响因子: 2.6
• 作者: Sim C;Denlinger DL
• 通讯作者: Denlinger DL

Molecular structure of the prothoracicotropic hormone gene in the northern house mosquito, Culex pipiens, and its expression analysis in association with diapause and blood feeding.

• DOI: 10.1111/j.1365-2583.2010.01058.x
• 发表时间: 2011-04
• 期刊: Insect molecular biology
• 影响因子: 2.6
• 作者: Zhang Q;Denlinger DL
• 通讯作者: Denlinger DL

Heat shock proteins contribute to mosquito dehydration tolerance.

• DOI: 10.1016/j.jinsphys.2009.09.012
• 发表时间: 2010-02
• 期刊: JOURNAL OF INSECT PHYSIOLOGY
• 影响因子: 2.2
• 作者: Benoit, Joshua B.;Lopez-Martinez, Giancarlo;Phillips, Zachary P.;Patrick, Kevin R.;Denlinger, David L.
• 通讯作者: Denlinger, David L.