NUCLEO-CYTOPLASMIC INTERACTIONS IN DEVELOPMENT

发育中的核质相互作用

• 批准号: 3314740
• 负责人: JULIE A DELOIA
• 金额: $ 17.88万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-08-01 至 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3314740
• 关键词: alleles; cell cell interaction; cell fusion; cell nucleus; cytogenetics; cytoplasm; developmental genetics; early embryonic stage; embryo /fetus culture; gametogenesis; gel electrophoresis; gene expression; genotype; gestational age; glucose 6 phosphate isomerase; immunofluorescence technique; laboratory mouse; lamins; lethal genes; mammalian embryology; oogenesis; phosphoglycerate kinase; sex chromosomes; sex linked trait; spermatogenesis; surface antigens; tissue mosaicism

We propose to investigate nucleo-cytoplasmic interactions which take place during normal development and to determine the mechanism by which these interactions are controlled. The role of various components of the mammalian zygote (male genome, female genome, egg cytoplasm) in development will be analyzed using recently developed nuclear transfer techniques. Our first major objective will be to determine the reasons for developmental failure of biparental androgenones (embryos with two male pronuclei) and biparental gynogenones (two female pronuclei). Development of chimeric embryos produced by aggregation of gynogenetic or androgenetic and normal embryos will be monitored in order to determine whether embryo- or cell-lethality is ultimately responsible for death of isoparental embryos. Haploid nuclei obtained from successive stages of male gametogenesis will be transferred into zygotes from which male nuclei were removed. These experiments will address the timing of appearance of functional differences between the male and the female genome. The second major objective will be to analyze development of enucleated zygotes into which nuclei from older developmental stages are transferred. We will determine if repeated nuclear transfer will improve the development of such embryos. The use of cytoplasm from older embryos (4- and 8-cell stage) as the nuclear recipient will be explored. Using 2D gel analysis we will monitor the changes in genome expression following nuclear transfer. Nuclear morphology, changes in nuclear proteins (lamins) and the structure of the mitotic apparatus in these nuclear transfer embryos will be investigated.

我们建议研究核质相互作用， 在正常发育期间并确定这些的机制 相互作用是受控的。 各组成部分的作用 发育中的哺乳动物受精卵（雄性基因组、雌性基因组、卵细胞质 将使用最近开发的核移植技术进行分析。 我们 第一个主要目标将是确定发展的原因， 双亲雄激素失败（胚胎有两个雄性原核）， 双亲雌核基因（两个雌性原核）。 嵌合体的开发 由雌核发育或雄核发育和正常的胚胎聚集而产生的胚胎 将对胚胎进行监测，以确定胚胎或 细胞致死性最终导致同系胚胎的死亡。 从雄性配子发生的连续阶段获得的单倍体细胞核将 被转移到受精卵中，从受精卵中去除雄性细胞核。 这些 实验将解决功能差异出现的时间 在男性和女性基因组之间。 第二个主要目标是 分析去核受精卵的发育， 发展阶段转移。 我们将确定是否重复 核移植将改善这些胚胎的发育。 使用 来自较老胚胎（4-和8-细胞期）的细胞质作为核受体 将被探索。 使用2D凝胶分析，我们将监测 核转移后的基因组表达。 核形态学，变化 核蛋白（核纤层蛋白）和有丝分裂器的结构， 将对这些核移植胚胎进行研究。