Lipid metabolism during tsetse reproduction

采采蝇繁殖过程中的脂质代谢

• 批准号: 8488404
• 负责人: Josh B. Benoit
• 金额: $ 1.35万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2013-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8488404
• 关键词: 1,2-diacylglycerol; Accounting; Adult; Africa; Africa South of the Sahara; African Trypanosomiasis; Animals; Antibiotics; Antibodies; Awareness; Bacteria; Biochemistry; Birth; Blood; Deposition; Development; Dietary Supplementation; Diglycerides; Disease; Disease Vectors; Economic Development; Environment; Fat Body; Fellowship; Female; Fertility; Food; Gene Expression; Gene Expression Profile; Gene Proteins; Generations; Genes; Genetic; Gland; Goals; Hemolymph; Human; Individual; Infertility; Ingestion; Insect Vectors; Insecta; Larva; Lipase; Lipid Mobilization; Lipids; Medical; Metabolism; Methods; Milk; Molecular; Molecular Biology; Mothers; Movement; Nutrient; Nutritional Requirements; Oogenesis; Orthologous Gene; Oviparity; Pathway interactions; Phenotype; Physiological; Physiology; Population; Population Sizes; Process; Protein Biosynthesis; Proteins; Public Health; RNA Interference; Reproduction; Reproductive Physiology; Role; Source; Sterility; Stimulus; Supplementation; System; Techniques; Training; Transcript; Triglycerides; Trypanosomiasis; Tsetse Flies; Vaccines; Wigglesworthia; Yeasts; adipokinetic hormone; aqueous; authority; base; disease transmission; egg; feeding; fly; health economics; insect disease; lipid metabolism; lipid transport; lipophorin; microbial; nagana; novel; offspring; peptide hormone; prevent; protein function; receptor; reproductive; transmission process; uptake; vector; vector control

DESCRIPTION (provided by applicant): The long-term objectives of the candidate (Dr. Joshua B. Benoit) are to become an authority on the physiology of insect disease vectors and increase public awareness of the role of insects on public health. The goal of this fellowship is to develop the candidate's ability to assess the biochemistry, molecular biology and physiology of vector reproduction. The training plan provides opportunities to conduct extensive studies at the interface between molecular biology and reproductive physiology. Tsetse flies are the sole vector of African sleeping sickness. Vaccine-based methods have not been successful for control of trypanosomiasis. A unique aspect of this disease vector is its K-strategy of reproduction. Mothers provide nutrients, directly to their intrauterine progeny via a milk gland and only give birth to 8-10 offspring in their lifetime. Interference with tsetse reproduction can supplement trapping and other control methods used to reduce tsetse population numbers and decrease disease transmission. Currently, there is little information on tsetse reproduction, particularly how nutrients are mobilized to the larva. Since this aspect is unique in tsetse, control techniques blocking maternal nutrients could be tsetse specific. The objectives of this study are to 1) characterize the metabolism and transfer of lipids from the mother to her offspring, and 2) decipher how this process is influenced by the presence of symbiotic bacteria. Characterization of lipid metabolism and mobilization will focus on the adipokinetic hormone (AKH; this portion has been completed) and Brummer (Bmm) lipase systems, as well as lipid transport by lipophorin (Lp) during the reproductive cycle. Gene expression and protein synthesis will be assessed spatially and temporally during larval intrauterine development. Function of these proteins will be assessed utilizing RNA interference to knockdown their expression. Role of symbiotic bacteria in lipid metabolism and movement during reproduction will be assessed by analyzing molecular and physiological differences between wild type and antibiotic-treated aposymbiotic flies (flies lacking symbionts). Aposymbiotic flies are infertile and abort undeveloped larva early in their developmental cycle. The mechanism(s) underlying this phenotype are unknown, but malnourishment due to milk secretions deficient in essential nutrients is the current hypothesis. A possible deficiency is in the lipid content of the milk. Differential lipid composition between wild type and aposymbiotic flies will be assessed by obtaining milk and comparing the quantitative and qualitative differences in milk lipids. Fertility can be restored in aposymbiotic flies by supplementation of blood meals with yeast extract. Lipid compositions of aposymbiotic flies with and without yeast extract-supplemented blood will also be performed. To determine genetic effects of dietary supplementation, wild-type and aposymbiotic flies will be compared by Illumina sequencing of whole body transcriptome. This project will provide avenues to develop proficiency in multiple molecular and physiological techniques that will provide the groundwork for the candidate to become an authority on the molecular physiology of disease vectors.

描述（由申请人提供）：候选人的长期目标（约书亚博士B。Benoit）将成为昆虫病媒生理学方面的权威，并提高公众对昆虫对公共卫生作用的认识。该奖学金的目标是培养候选人评估载体繁殖的生物化学、分子生物学和生理学的能力。培训计划提供了机会，在分子生物学和生殖生理学之间的接口进行广泛的研究。采采蝇是非洲昏睡病的唯一传播媒介。基于疫苗的方法尚未成功控制锥虫病。这种疾病载体的一个独特方面是其繁殖的K策略。母亲通过乳腺直接向胎儿提供营养，一生只生8-10个后代。干扰采采蝇繁殖可以补充诱捕和其他控制方法，以减少采采蝇种群数量和减少疾病传播。目前，关于采采蝇繁殖的信息很少，特别是营养物质如何被动员到幼虫中。由于这方面在采采蝇中是独特的，阻断母体营养的控制技术可能是采采蝇特有的。本研究的目的是1）表征脂质从母体到后代的代谢和转移，2）破译共生细菌的存在如何影响这一过程。脂质代谢和动员的表征将集中在脂肪动力激素（AKH;这一部分已经完成）和布鲁默（Bmm）脂肪酶系统，以及在生殖周期中通过脂转运蛋白（Lp）的脂质转运。基因表达和蛋白质合成将评估空间和时间在幼虫宫内发育。这些蛋白质的功能将利用RNA干扰敲低其表达来评估。共生细菌在脂质代谢和运动在繁殖过程中的作用将通过分析野生型和驱虫处理的非共生苍蝇（苍蝇缺乏共生体）之间的分子和生理差异进行评估。非共生蝇是不育的，在发育周期的早期会使未发育的幼虫流产。这种表型的潜在机制尚不清楚，但目前的假设是由于乳汁分泌物缺乏必需营养素而导致营养不良。一个可能的缺陷是牛奶中的脂质含量。将通过获得乳汁并比较乳汁脂质的定量和定性差异来评估野生型和非共生果蝇之间的脂质组成差异。通过在血粉中补充酵母提取物，可以恢复非共生果蝇的生育能力。还将进行有和没有酵母提取物补充血液的非共生苍蝇的脂质组成。为了确定膳食补充剂的遗传效应，将通过全身转录组的Illumina测序来比较野生型和非共生果蝇。该项目将提供途径，发展多种分子和生理技术的熟练程度，这将为候选人成为疾病载体的分子生理学权威奠定基础。