Expanding the toolbox for tsetse reproductive biology

描述（由申请人提供）：采采蝇传播人类和动物形式的非洲锥虫病，这是一种被忽视的疾病，影响撒哈拉以南非洲的边缘化人群。基于农药诱饵靶标和诱捕器的控制方法可以减少采采蝇种群，但具有缺点，包括环境毒性、对非靶标生物的附带影响和高实施成本。因此，需要基于遗传和非农药的病媒控制方法。该项目将为旨在降低采采蝇生殖能力的控制方法奠定基础。我们将研究雄性附腺（MAG）的产物，即精液蛋白（SFP），它在交配过程中随精子沿着转移到雌性体内。SFPs介导交配后雌性昆虫重要的生理和行为变化，包括降低再交配的接受性和增加卵子发生/卵子沉积。破坏/操纵SFP功能有可能减少采采蝇种群，从而减少疾病传播。采采蝇的生殖能力很低，因为它们通过专性胎生繁殖。在每个生殖营养周期，雌性在发育期间携带并养育一个宫内后代。雌性平均一生生育8-10个后代。我们提出的研究得到了以下可用性的支持：1）我们的目标物种Glossina morsitans morsitans（Gmm）中MAG基因表达的初步转录组数据，2）Gmm的注释基因组和广泛的转录组资源，以及3）来自其他采采蝇物种的转录组的大型数据集。本研究有3个目的：1。使用转录组学，蛋白质组学和基因组学方法确定推定的SFP基因，表征其性别/组织特异性，并在成熟过程中和交配后检查时间表达谱。2.利用现有的转录组学和基因组学资源鉴定多个Glossina属物种之间的SFP直系同源物和旁系同源物，通过多态性检测估计SFP的序列多样性，并通过计算机分析鉴定保守的性别和组织特异性调控元件。3.开发和应用基因沉默技术，以评估选定的SFP（~5）在采采蝇生殖适合度参数范围内的功能作用。

DESCRIPTION (provided by applicant): 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.