METABOLIC CHANGES IN PREIMPLANTATION EMBRYOS

植入前胚胎的代谢变化

• 批准号: 3314429
• 负责人: HARRY M WEITLAUF
• 金额: $ 13.74万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3314429
• 关键词: antibody specificity; dissection; early embryonic stage; embryo /fetus culture; embryo /fetus protein; embryo implantation; endometrium; estrogens; gel electrophoresis; genetic transcription; hormone regulation /control mechanism; immunoprecipitation; laboratory mouse; mammalian embryology; messenger RNA; protein biosynthesis

This project is concerned with the regulation of metabolism in preimplantation mouse embryos. The overall objective is to define the mechanism by which the mother can render delayed implanting embryos dormant, maintain them in that condition for several days, and then cause them to resume development, and implant. Experiments to be conducted in the upcoming grant period will focus on the regulation and function of a serieso f proteins that are synthesized and secreted by the embryo in the immediate preimplantation period. Synthesis of some of these proteins appears to be regulated in response to the signals that terminate the embryonic diapause associated with delayed implantation; others appear to be constitutive. Dormant embryos will be incubated with (3H) methionine and those undergoing reactivation will be incubated with (35S) methionine; the proteins that are synthesized and secreted will be mixed together and analyzed by 2-D electrophoresis on polyacrylamide gels under denaturing conditions. The proteins will be localized on the gels by fluorography or autoradiography and the ratio of (3H) to (35S) in the spots of interest will be determined with a scintillation counter. This ratio will be normalized to the (3H)/(35S) for overall protein synthesis in the same embryos to provide an estimate of the relative change in synthesis of each protein of interest. In this way it is hoped to determine whether synthesis of these proteins is regulated differentially. In addition, the temporal relationship of the appearance of these embryonic proteins to implantation-associated changes in the endometrium will be determined. Finally, experiments will be conducted to evaluate the possibility that the proteins secreted by mouse blastocysts interact with the endometrium by binding to the plasma membrane and thus may act as an embryonic signaling factor as has been shown to be the case for the protein (oTP-1) that is secreted by sheep blastocysts.

该项目涉及对代谢的调节 植入前小鼠胚胎。 总体目标是 定义母亲可以延迟的机制 植入休眠的胚胎，将其保持在这种情况下 几天，然后使他们恢复发展，并且 注入。 在即将到来的赠款中进行的实验 周期将重点放在系列F的调节和功能上 由胚胎合成和分泌的蛋白质 立即植入期。 其中一些的综合 蛋白质似乎是根据信号受到调节的 终止与延迟相关的胚胎滞育 植入；其他似乎是构成的。 休眠胚胎 将与（3H）甲硫氨酸孵育 重新激活将与（35s）蛋氨酸孵育；蛋白质 合成和分泌的这些将混合在一起， 通过2-D电泳在聚丙烯酰胺凝胶上分析 变性条件。 蛋白质将位于凝胶上 通过荧光或放射自显影以及（3H）与（35s）的比率 在感兴趣的地方将用闪烁确定 柜台。 该比率将标准化为（3h）/（35s） 在同一胚胎中的总体蛋白质合成以提供 估计每种蛋白质合成的相对变化 兴趣。 通过这种方式，希望确定是否合成 这些蛋白质受到差异调节。 另外， 这些胚胎外观的时间关系 子宫内膜的蛋白质与植入相关的变化 将确定。 最后，将进行实验 评估小鼠分泌的蛋白质的可能性 胚泡通过与子宫内膜相互作用 质膜，因此可以充当胚胎信号传导 因子已证明是蛋白质（OTP-1）的情况 这是由绵羊胚泡分泌的。