CELL LINEAGES IN MAMMALIAN DEVELOPMENT

含有两个父本（双亲andogenones）的小鼠胚胎 而两个母本（双亲雌核母核）原核不 完全发育，但在植入后不久死亡。 这些胚胎的植入前发育似乎 形态正常。 雄核发育胚胎表现出 胚胎发育正常，而雌核发育失败， 胚胎外细胞生长和/或分化不良 膜。 本提案的总体目标是分析 来自雌激素、雄激素和 聚集嵌合体中的正常胚胎，从而确定 无论是细胞致死还是形态发生相互作用的失败， 导致发育失败。 我们建议建造 聚集嵌合体使用雄激素的各种组合， 雌核发育和正常卵裂球。 后续发展 这些胚胎将被监测，每个胚胎的参与 成分确定。 为了确定 在这种嵌合胚胎中的每个细胞的原位衍生 将使用识别方法。 我们建议使用胚胎 从我们实验室已经开发的转基因小鼠品系中。 外源DNA序列将在每个细胞的细胞核中被检测到， 使用生物素化探针的原位DNA杂交。 由于每个 来自转基因胚胎的细胞携带特异性可检测的 序列，每个组成部分的起源和贡献， 可以容易地确定嵌合胚胎。 通过比较空间 雌核发育、雄核发育和 胚胎内的正常细胞，具有以下发育能力： 各种嵌合体胚胎，我们将确定父系的作用， 和母体基因组在小鼠早期形态发生中的作用 发展

Mouse embryos containing two paternal (biparental andogenones) and two maternal (biparental gynogenones) pronuclei do not complete development but die soon after implantation. Preimplantation development of such embryos appears morphologically normal. Androgenetic embryos display failure of development of the embryo proper while gynogenones fail because of poor growth and/or differentiation of extraembryonic membranes. The overall objective of this proposal is to analyze the interactions of cells from gynogenones, androgenones and normal embryos in aggregation chimeras, thus determining whether cell lethality or failure of morphogenetic interactions is responsible for developmental failure. We propose to construct aggregation chimeras using various combinations of androgenetic, gynogenetic and normal blastomeres. Subsequent development of such embryos will be monitored and participation of each component determined. In order to determine the exact derivation of each cell in such chimeric embryos, in situ identification methods will be used. We propose to use embryos from transgenic mouse lines already developed in our laboratory. Foreign DNA sequences will be detected in nucleus of each cell by in situ DNA hybridization using biotinylated probes. Since each cell derived from transgenic embryos carries specific detectable sequences, the origin and contribution of each component of the chimeric embryo can be easily determined. By comparing spatial and quantitative distribution of gynogenetic, androgenetic and normal cells within embryos with the developmental capacity of various chimeric embryos we will determine the role of paternal and maternal genome in early morphogenetic events during mouse development.