Massively parallel sequencing of the diapause transcriptome in Aedes albopictus

描述(申请人提供)：亚洲虎蚊子，白纹伊蚊(Skuse)，是登革热的重要媒介，也有效地传播黄热病、基孔肯雅病毒和几种北美本土虫媒病毒。在过去20年里，这种蚊子从其原生的亚洲地区迅速传播到全球各地，最近参与了疾病暴发，这是一个突出的公共卫生问题。光周期介导的卵滞育是白纹伊蚊和其他与医学相关的蚊子在温带环境中生存并在广泛地理范围内传播的关键生态特征。这一研究计划的长期目标是严格定义白纹伊蚊光周期滞育的分子基础，作为基于破坏这种关键的生态适应而开发新的媒介控制形式的基础。这项建议的直接目标是确定白纹伊蚊光周期滞育反应的转录元件。在特定的目标一，大规模平行的焦磷酸测序和生物信息学分析将用于比较来自卵母细胞、发育中的胚胎和海鞘幼虫的滞育和非滞育转录本。在具体目标2中，将通过比较滞育(温带)和非滞育(热带)种群的基因表达，利用定量RT-PCR来识别支持滞育反应的转录事件。除了为开发新的媒介控制形式提供基础外，这些研究的结果还将迅速推进蚊子应激反应生理学的基因发现和理解。与公共卫生相关的光周期滞育是指昆虫测量日长(光周期)的能力，以此作为启动季节性适当的发育停滞模式的线索。本研究将探讨白纹伊蚊对光周期滞育的分子控制。这项研究的结果将为在破坏这一关键的生态适应的基础上开发新的媒介控制策略提供基础。

DESCRIPTION (provided by applicant): The Asian tiger mosquito, Aedes albopictus (Skuse), is an important vector of dengue fever and also efficiently transmits yellow fever, chikungunya virus and several native North American arboviruses. The rapid spread of this mosquito from its native Asian range across the globe during the last 20 years and its recent involvement in disease outbreaks represents an outstanding public health concern. Photoperiodically mediated egg diapause is a pivotal ecological trait central to the ability of A. albopictus and other medically relevant mosquitoes to inhabit temperate environments and spread across broad geographic ranges. The long-term goal of this research program is to rigorously define the molecular basis of photoperiodic diapause in A. albopictus as a foundation for developing novel forms of vector control based on the disruption of this critical ecological adaptation. The immediate goal of this proposal is to identify transcriptional elements of the photoperiodic diapause response in A. albopictus. In Specific Aim One, massively parallel pyrosequencing and bioinformatics analyses will be used to compare diapause vs. non-diapause transcriptomes from oocytes, developing embryos, and pharate larvae. In Specific Aim 2, quantitative RT- PCR will be employed to identify transcriptional events underpinning the diapause response by comparing gene expression between a diapausing (temperate) and non- diapausing (tropical) population. In addition to providing a foundation for developing novel forms of vector control, results from these studies will rapidly advance gene discovery and understanding of stress response physiology in mosquitoes. PUBLIC HEALTH RELEVANCE Photoperiodic diapause refers to the ability of insects to measure day length (photoperiod) as a cue for initiating seasonally appropriate patterns of developmental arrest. This study will investigate the molecular control of photoperiodic diapause in the invasive and medically important mosquito, Aedes albopictus. Results of this study will provide a foundation for developing novel strategies for vector control based on the disruption of this critical ecological adaptation.