First Transcriptome to the Tick Olfactory System

蜱嗅觉系统的第一个转录组

基本信息

批准号：
8305997
负责人：
Richard Michael Roe
金额：
$ 18.66万
依托单位：
NORTH CAROLINA STATE UNIVERSITY RALEIGH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8305997
关键词：
Adult American Animal Diseases Animal Model Ankle Arthropods Behavior Biological Assay Bite Black-legged Tick Blood Boxing Canis familiaris Communication Culicidae Deet Dermacentor Detection Development Developmental Biology Elements Female Future Gene Expression Gene Expression Profile Goals Health Indium Insect Vectors Insecta Knowledge Leg Methods Microarray Analysis Molecular Molecular Biology Nature Odors Organ Paper Partner in relationship Pheromone Physiology Play Preparation Process Protein Databases Proteins Publishing RNA RNA Interference Reproduction Research Rickettsia rickettsii Risk Rocky Mountain Spotted Fever Role Screening procedure Smell Perception Staging Structure Study models System Ticks Time Tissues United States Vector-transmitted infectious disease Work base disease transmission feeding human disease infancy insect disease interest male next generation odorant-binding protein receptor remediation response soft tissue transcriptomics vector

项目摘要

DESCRIPTION (provided by applicant): Ticks are major vectors of human and animal diseases and a significant nuisance problem. The American dog tick, Dermacentor variabilis, which is the focus of this proposal is the vector of Rocky Mountain spotted fever in the US, and an excellent model for the studies proposed, in part, because of its large size, ease of rearing and the background research on its development. Critical to disease transmission is host recognition, blood feeding, pheromone detection, mating and reproduction. Also important to minimizing host contact with ticks, is the use of personal repellents. It has long been recognized that olfaction is a critical aspect of these processes, and extensive studies have been conducted on olfaction including functional transcriptomics in other blood feeding arthropods like mosquitoes. However, essentially nothing is known about olfaction at the molecular level in ticks. The alternative to the insect antenna in ticks is the Haller's organ, a unique structure found on the tarsi on the first pair of legs. The Haller's organ is basically a molecular black box, and no transcriptome to this organ is available. In preliminary studies in preparation for this proposal, we have developed transcriptomes and successfully examine global gene expression for the first time in the Dv synganglion during female adult reproduction, developed bioassay methods for RNAi screening in the American dog tick, and optimized methods for RNA extraction from tarsi of Dv. The tarsi consist mostly of hard cuticle with minimal soft tissue, and RNA extraction in high yield with good quality is difficult. We have also developed a new, commercialized (natural) tick repellent, BioUD (more effective than Deet) and are interested in understanding the mechanism of repellency and developing rationale approaches for future repellent discovery. Dr. Sonenshine, a co-PI on this proposal, has also conducted much of the pioneering work on tick pheromones and their role in adult reproduction in Dv. The current proposal is to develop the first transcriptome to a tick Haller's organ and to conduct microarray analysis and RNAi screening to identify targets (odorant binding proteins, receptors, catabolic systems and other proteins) for future mechanistic studies of tick olfaction and the development of next generation control strategies for ticks. This will be achieved by the following approaches: Objective 1: Construct the first 454 transcriptome to the Haller's organ of ticks (the model organism will be Dv) and assign putative functions to the assembled contigs; Objective 2: Develop an Agilent Haller's organ microarray and examine global gene expression in response to tick-host interactions and adult reproduction; and Objective 3: Conduct RNAi screening to identify critical messages important in tick olfaction and development. We expect the work proposed will greatly increase our knowledge of the molecular biology of tick olfaction which is currently in its infancy and stimulate future work on chemoreception in this important arthropod group. Future research will include gustatory receptors in the tick palps and their function in tick development and repellency.

描述（由申请人提供）：壁虱是人类和动物疾病的主要媒介，也是一个重大的滋扰问题。美国狗的tick tick虫，Dermacentor variabilis，这是该提案的重点是美国落基山斑点发烧的向量，这是提议的一部分研究，部分原因是它的较大尺寸，易于饲养，易于饲养和背景研究其发展。疾病传播至关重要的是宿主识别，血液喂养，信息素检测，交配和繁殖。最大程度地降低了与tick的接触，这也是个人驱虫剂的使用。长期以来，人们已经认识到，嗅觉是这些过程的关键方面，并且已经对嗅觉进行了广泛的研究，包括在其他血液喂养节肢动物（如蚊子）中的功能转录组学。但是，从本质上讲，关于滴答滴答中分子水平的嗅觉一无所知。 tick中昆虫天线的替代方法是Haller的器官，这是第一对腿上的Tarsi上发现的独特结构。 Haller的器官基本上是一个分子黑匣子，没有转录到该器官。在为该提案做准备的初步研究中，我们在女性成人繁殖过程中首次开发了转录组，并在DV Synganglion中首次成功检查了全球基因表达，开发了用于美国狗tick中RNAi筛查的生物测定方法，并针对RNA提取方法进行了RNA提取方法来自DV的Tarsi。 TARSI主要由硬性角质层组成，其软组织最少，而RNA提取的质量很高。我们还开发了一种新的，商业化的（自然）tick驱虫，Bioud（比DEET更有效），并有兴趣了解驱虫的机制，并为未来的驱虫剂开发了理由方法。这项提案的共同PI Sonenshine博士还进行了许多关于tick信息素的开创性工作及其在DV中成人复制中的作用。当前的建议是将第一个转录组发展为tick Haller的器官，并进行微阵列分析和RNAi筛选，以鉴定靶标（气味结合蛋白，受体，分解代谢系统和其他蛋白质），以使tick嗅觉的未来机械研究以及下一步的开发壁虱的生成控制策略。这将通过以下方法来实现：目标1：将第一个454个转录组构建到Haller的tick器器官（模型有机体为DV），并将假定的功能分配给组装的重叠群；目标2：开发一个敏捷的Haller的器官微阵列，并检查全球基因表达，以响应tick-Host的相互作用和成人繁殖；和目标3：进行RNAi筛选，以确定在壁虱嗅觉和开发中重要的关键信息。我们预计所提出的工作将大大增加我们对tick嗅觉的分子生物学的了解，这目前尚处于起步阶段，并激发了这个重要的节肢动物组中未来对化学感受的工作。未来的研究将包括tick板中的味觉受体及其在tick开发和驱逐心中的功能。