INDUCTION OF AMINO ACID TRANSPORT SYSTEMS XC, GLY AND XA

氨基酸转运系统 XC、GLY 和 XA 的诱导

• 批准号: 2204263
• 负责人: DAVID F MANN
• 金额: $ 10.36万
• 依托单位: MIDWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-06-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2204263
• 关键词: aminoacid metabolism; aminoacid transport; egg /ovum; fertilization; glutathione; graafian follicles; high performance liquid chromatography; laboratory mouse; ovulation; oxidative stress; protein biosynthesis; tissue /cell culture

The overall goal of this project is to study the induction of the amino acid transport systems specific for the precursors of glutathione in the mouse ovarian follicle and in oocytes. Our preliminary studies show that the amino acid transport systems for the three precursors of glutathione, cystine, glycine and glutamate, are induced in preantral follicles after being cultured four hours in Waymouth medium in air containing 5% carbon dioxide [20% oxygen]. The specific aims are: (l) to identify the induced Na-independent and Na-dependent glutamate uptake systems; (2) to determine the factors required for the induction of amino acid transport systems for cystine, glycine and glutamate and the role of uptake on glutathione synthesis; (3) to test the hypothesis that ovulation and fertilization are in vivo events that result in the induction of systems x(c-) and Gly. Three factors, amino acids, oxidant stress and glutathione, will be investigated as potential candidates regulating the induction of systems x(c-), X(AG-) and Gly and glutathione levels. In amino acid supplementation experiments, preantral follicles will be cultured with selected amino acids at 3% oxygen to minimize oxidant stress. When follicles are cultured with different levels of oxygen, superoxide anion and hydrogen peroxide, Waymouth medium will be supplemented with the amino acids needed for glutathione synthesis. When varying glutathione levels, follicles will be cultured at 3% oxygen and in amino acid supplemented medium. Timing of the in vivo expression of systems x(c-) and Gly and glutathione levels will be determined in ovulated-unfertilized and fertilized oocytes. These studies may benefit work involving in vitro culture of immature oocytes and fertilization of mature oocytes in agriculturally important species.

该项目的总体目标是研究氨基的诱导 酸转运系统特有的是谷胱甘肽的前体 小鼠卵巢卵泡和卵母细胞。 我们的初步研究表明 谷胱甘肽三个前体的氨基酸传输系统， 胱氨酸，甘氨酸和谷氨酸在酸中诱导 在含5％碳的空气中的Waymouth培养基中培养四个小时 二氧化物[20％氧]。 具体目的是：（l）识别诱导的 非NA独立和NA依赖性的谷氨酸摄取系统； （2）确定 诱导氨基酸传输系统所需的因素 胱氨酸，甘氨酸和谷氨酸以及摄取在谷胱甘肽上的作用 合成; （3）检验排卵和施肥的假设是 体内事件导致系统X（C-）和Gly的诱导。 三个因素，氨基酸，氧化剂应激和谷胱甘肽，将是 被调查为调节系统诱导的潜在候选者 x（c-），x（ag-）和gly和谷胱甘肽水平。 在氨基酸中 补充实验，尿布卵泡将与 在3％的氧气下选择的氨基酸以最大程度地减少氧化应激。 什么时候 卵泡以不同水平的氧气，超氧化物阴离子培养 和过氧化氢，Waymouth培养基将补充氨基 谷胱甘肽合成所需的酸。 当改变谷胱甘肽水平时， 卵泡将以3％的氧气培养，并在氨基酸中培养 中等的。 系统x（c-）和gly的体内表达的时机 谷胱甘肽水平将在排卵的非饮用物中确定，并确定 受精的卵母细胞。 这些研究可能有益于涉及体外的工作 未成熟卵母细胞的培养和成熟卵母细胞的受精 农业重要的物种。