Expanding the toolbox for tsetse reproductive biology

项目摘要/摘要 采采蝇传播非洲锥虫病的人和动物形式，被忽视的疾病，影响 撒哈拉以南非洲的边缘化人口。基于农药投饵靶标和诱捕器的防治方法 可以减少采采量，但也有负面影响，包括环境毒性、对非 目标生物和高实施成本。因此，以遗传和非农药为基础的媒介生物控制方法 都是可取的。该项目将为旨在减少采采蝇繁殖的控制方法奠定基础 容量。我们将研究男性附腺(MAGS)产品，即精液蛋白(SFP)， 在交配过程中，它们与精子一起传递给雌性。SFP介导重要的生理和 交配雌虫的行为变化，包括对交配的接受性降低和增加 卵子发生/卵子沉积。SFP功能的中断/操作(S)有可能减少采采率 人口，因此疾病传播。采采蝇的繁殖能力很低，因为它们通过 义务胎生。在每个性腺营养周期中，雌性携带并滋养一只宫内后代 其发展的持续时间。雌性平均每一生产下8-10只后代。我们建议的研究是 1)MAG基因在我们的目标物种中表达的初步转录数据的可用性支持 Glossina morsitans morsitans(GMM)，2)GMM的注释基因组和广泛的转录资源， 3)其他采采族物种的转录本的大型数据集。这项研究有三个目的： 1.利用转录组、蛋白质组和基因组学方法鉴定可能的SFP基因，鉴定其特征 性别/组织特异性，并检查成熟期间和交配后的时间表达谱。 2.利用已有的转录切割法确定多个舌目物种之间的SFP同源基因和同源基因 和基因组资源，通过多态检测和鉴定来评估SFP的序列多样性 在电子计算机分析中保守性别和组织特异性调控元件。 3.开发和应用基因沉默技术来评估选定的SFP(~5)在体内的功能作用(S 采采子生殖适合度参数的上下文。

Project Summary/Abstract Tsetse flies transmit the human and animal forms of African Trypanosomiasis, neglected diseases, which affect marginalized populations in sub-Saharan Africa. Control methods based upon pesticide-baited targets and traps can reduce tsetse populations, but have downsides, including environmental toxicity, collateral effects upon non- target organisms and high implementation cost. Thus, genetic and non-pesticide based vector control methods are desirable. This project will build the foundation for control methods that aim to reduce tsetse's reproductive capacity. We will investigate male accessory gland (MAGs) products, known as seminal fluid proteins (SFPs), which are transferred along with sperm to the female during mating. SFPs mediate important physiological and behavioral changes in mated female insects, including reduced receptivity to remating and increased oogenesis/egg deposition. Disruption/manipulation of SFP function(s) has the potential to reduce tsetse populations and hence disease transmission. Tsetse flies have a low reproductive capacity as they reproduce by obligate viviparity. Each gonotrophic cycle, females carry and nourish a single intrauterine offspring for the duration of its development. Females on average produce 8-10 offspring per lifetime. Our proposed studies are supported by the availability of 1) preliminary transcriptomic data on MAG gene expression in our target species Glossina morsitans morsitans (Gmm), 2) an annotated genome and extensive transcriptomic resources for Gmm, and 3) a large dataset of transcriptomes from other tsetse species. This study has 3 aims: 1. Identify putative SFP genes using transcriptomic, proteomic and genomic approaches, characterize their sex/tissue specificity, and examine temporal expression profiles during maturation and after mating. 2. Identify SFP orthologs and paralogs between multiple Glossina species by utilizing available transcriptomic and genomic resources, estimate sequence diversity of SFPs by polymorphism detection and identify conserved sex and tissue specific regulatory elements by in silico analysis. 3. Develop and apply gene silencing techniques to assess the functional role(s) of selected SFPs (~5) within the context of the tsetse reproductive fitness parameters.