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（Joshua B. Benoit博士）将成为昆虫疾病媒介生理学的权威，并提高公众对昆虫在公共卫生中的作用的认识。该团契的目的是发展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。培训计划提供了在分子生物学和生殖生理学之间接口进行广泛研究的机会。采摘苍蝇是非洲昏睡病的唯一媒介。基于疫苗的方法尚未成功控制锥虫病。该疾病载体的一个独特方面是其繁殖的k-strategy。母亲通过牛奶腺直接向宫内后代提供营养，只生下了8-10个后代。干扰采集采集的繁殖可以补充诱捕和其他用于减少采采群体数量并减少疾病传播的控制方法。当前，关于采集繁殖的信息很少，尤其是养分如何动员到幼虫。由于这方面在采集中是独一无二的，因此控制技术阻止母体营养可能是采集特定的。这项研究的目标是1）表征脂质从母亲到她的后代的代谢和转移，以及2）解读该过程如何受共生细菌的存在影响。脂质代谢和动员的表征将集中于脂肪激素（AKH；此部分已经完成）和BRUMMER（BMM）脂肪酶系统，以及在生殖循环期间通过脂蛋白（LP）通过脂质转运。基因表达和蛋白质合成将在幼虫内发育期间在空间和时间上进行评估。这些蛋白质的功能将利用RNA干扰来评估其表达。共生细菌在脂质代谢和繁殖过程中的运动中的作用将通过分析野生型和抗生素治疗的阿物蛋白无生物生物蝇之间的分子和生理差异来评估（缺乏共生体）。阿司生物蝇是不育的，并且在其发育周期初期流产未开发的幼虫。该表型的基础机制尚不清楚，但是由于牛奶分泌不足而营养不良，这是当前的假设。可能的缺乏是牛奶的脂质含量。将通过获得牛奶并比较牛奶脂质的定量和定性差异来评估野生型和末期蝇之间的差异脂质组成。通过用酵母提取物补充血液粉，可以在凋亡果蝇中恢复生育能力。还将进行伴有有或没有酵母提取物补充血液的凋亡蝇的脂质组成。为了确定补充饮食的遗传作用，将通过整个体转录组的光照明测序比较野生型和凋亡的苍蝇。该项目将为发展多种分子和生理技术的水平提供途径，这将为候选人成为疾病媒介分子生理的权威提供基础。