UPTAKE OF LIPIDS BY INSECT OOCYTES

昆虫卵母细胞对脂质的吸收

• 批准号: 2188463
• 负责人: HENDRICUS G VAN ANTWERPEN
• 金额: $ 10.12万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2188463
• 关键词: Manduca; SDS polyacrylamide gel electrophoresis; active sites; binding proteins; cofactor; complementary DNA; electron microscopy; enzyme activity; enzyme biosynthesis; gel filtration chromatography; high performance liquid chromatography; immunocytochemistry; immunofluorescence technique; in situ hybridization; lipid metabolism; lipid transport; lipoprotein lipase; messenger RNA; molecular cloning; northern blottings; nucleic acid sequence; oogenesis

The long-term objective of the proposed project is to understand the mechanisms that govern the uptake of lipids by insect oocytes. This will fill a gap in our knowledge on insect development, which may lead to the design of novel and effective ways to control the insects that play a role in the transmission of diseases like malaria, yellow fever, river blindness, Chagas disease, etc. The proposed project will focus on a recently identified lipoprotein lipase from mature eggs of the hawkmoth Manduca sexta. The observed hydrolysis of lipoprotein-transported diacylglycerol by this lipoprotein lipase indicate a critical role of the enzyme in the uptake of lipids by the developing oocyte. The specific aims of the project are: 1. To determine the biochemical characteristics of the lipoprotein lipase. Using standardized lipoprotein lipase assays, various lipid substrates and inhibitors of lipase activity, the enzyme will be characterized and compared with known vertebrate lipases. 2. To determine the site of synthesis and the subcellular site of activity of the lipase. Analyzing the synthesis of lipoprotein lipase and its mRNA in different tissues, combined with histological and immunocytochemical localization of the enzyme, will reveal sites at which the uptake of lipids by the oocyte is controlled. 3. To develop molecular probes of the lipoprotein lipase. Molecular cloning and sequencing of lipoprotein lipase cDNA will reveal the degree of homology with vertebrate lipases and will provide valuable probes for developmental studies. 4. To determine the possible role of other protein factors in lipid transport to the oocyte. In addition to the role of lipoprotein lipase, the possible involvement and relative importance of putative lipoprotein receptors, lipid transfer proteins and fatty acid binding proteins in the uptake of lipids by insect oocytes will be assessed.

拟议项目的长期目标是了解 昆虫卵母细胞摄取脂质的机制。这 将填补我们对昆虫开发的知识的空白，这可能会导致 设计新颖有效的方法来控制昆虫 在疟疾，黄热病等疾病的传播中发挥作用 河流失明，查加斯病等。 拟议的项目将重点放在最近确定的脂蛋白上 霍克莫斯·曼达卡·塞克斯塔（Hawkmoth Manduca Sexta）成熟卵的脂肪酶。观察到 通过这种水解脂蛋白转运的二酰基甘油 脂蛋白脂肪酶表明该酶在摄取中的关键作用 发育中的卵母细胞的脂质。项目的具体目的 是： 1。确定脂蛋白的生化特征 脂肪酶。使用标准化的脂蛋白脂肪酶测定，各种脂质 脂肪酶活性的底物和抑制剂，酶将是 表征并与已知的脊椎动物脂肪酶进行比较。 2。确定合成部位和亚细胞位点 脂肪酶的活性。分析脂蛋白脂肪酶的合成 及其在不同组织中的mRNA，结合了组织学和 酶的免疫细胞化学定位将揭示位点 卵母细胞对脂质的摄取受到控制。 3。开发脂蛋白脂肪酶的分子探针。分子 脂蛋白脂肪酶cDNA的克隆和测序将揭示 与脊椎动物脂肪酶的同源程度，将提供有价值的 发展研究的探针。 4。确定其他蛋白质因子在脂质中的可能作用 运输到卵母细胞。除了脂蛋白脂肪酶的作用外， 推定的可能参与和相对重要性 脂蛋白受体，脂质转移蛋白和脂肪酸结合 将评估昆虫卵母细胞摄入脂质中的蛋白质。