Expanding the toolbox for tsetse reproductive biology

描述(申请人提供)：采采蝇传播非洲锥虫病的人和动物形式，这是一种被忽视的疾病，影响撒哈拉以南非洲的边缘人群。基于农药靶标和诱捕器的防治方法可以减少采采蝇种群，但存在环境毒性、对非靶标生物的附带影响和较高的实施成本等缺点。因此，基因和非农药为基础的媒介生物控制方法是可取的。该项目将为旨在降低采采树繁殖力的控制方法奠定基础。我们将研究雄性附腺(MAGS)产物，即精液蛋白(SFP)，它们在交配过程中与精子一起传递给雌性。SFP介导交配后雌性昆虫的重要生理和行为变化，包括降低对交配的感受性和增加卵子发生/卵沉积。干扰/操纵SFP功能(S)有可能减少采采症种群，从而减少疾病传播。采采蝇的繁殖能力很低，因为它们通过预定的胎生繁殖。在每个性腺营养周期中，雌性在其发育期间孕育和滋养一只宫内后代。雌性平均每一生产下8-10只后代。我们提出的研究得到了以下方面的支持：1)关于MAG基因在我们的目标物种Glossina morsitans morsitans(GMM)中表达的初步转录数据，2)GMM的注释基因组和广泛的转录转录资源，以及3)其他采采族物种的大量转录数据集。本研究有三个目的：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，鉴定它们的性别/组织特异性，并检测成熟和交配后的时间表达谱。2.利用现有的转录和基因组资源，鉴定不同物种间的SFP同源基因和同源基因，通过多态检测估计SFP的序列多样性，通过电子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，在采摘繁殖适合度参数的背景下，评估选定的SFP(~5)的功能作用(S)。

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.