ORIGIN OF TRANSGENE-RELATED OVARIAN TERATOCARCINOMAS

描述(改编自《调查者摘要》)：这是一个新的 申请四年资助研究转基因小鼠 TG.KD系，卵巢恶性发生率高 处于半合子状态的畸胎癌。卵巢畸胎瘤是一种 起源于女性生殖细胞。它们是一种常见的，但知之甚少的 从多潜能干细胞发展而来的女性肿瘤。这根茎 细胞，胚胎癌细胞(EC)，可能是由表观遗传学产生的 来自孤雌生殖激活的生殖细胞的机制，并可以 分化成各种各样的胚胎组织，形成畸胎瘤。一个 畸胎癌是一种恶性的畸胎瘤，它包含 未分化的EC细胞以及已分化的衍生品。这个 拟议研究的主要目标是分离突变基因 TG.KD转基因插入位点和了解TG.KD的作用 卵巢畸胎癌发生发展中的分离基因。三 提出了具体目标： 1.检验卵巢畸胎癌发生于TG.KD的假说 作为转基因插入失活或激活结果的品系 在宿主基因中，转基因整合位点的转录本将是 鉴定、克隆和表征。 2.为了评估在生殖细胞发育过程中突变基因的作用时间，F1 将携带TG.KD的FVB/N自交系与 其他自交系。由此产生的畸胎癌将使用以下方法进行基因分型 几个种的着丝粒周围和远端区域的异形标记 染色体。标记相对F1体细胞的分布 细胞将被用来确定减数分裂的最新阶段 畸胎癌的形成。 3.检测TG.KD转基因纯合子小鼠的表型 插入。虽然半合子TG.KD转基因小鼠有倾向于 发生卵巢畸胎癌，纯合子小鼠表现出围产期 杀伤力。基于工作假设，相同的基因突变 导致畸胎癌和围产期死亡的原因是 将对纯合子胚胎的表型进行组织学检查 提供有关突变基因功能的信息。

DESCRIPTION (Adapted from the Investigator's Abstract): This is a new application requesting four years of funding to study a transgenic mouse line, TG.KD, which has a high incidence of malignant ovarian teratocarcinomas in the hemizygous state. Ovarian teratomas are tumors that originate from female germ cells. They are a common, yet poorly understood neoplasm of women that develops from pluripotential stem cells. The stem cells, embryonal carcinoma (EC) cells, probably arise by an epigenetic mechanism from parthenogenetically activated germ cells, and can differentiate into a wide variety of embryonic tissues to form teratomas. A teratocarcinoma is a malignant form of teratoma that contains undifferentiated EC cells in addition to differentiated derivatives. The primary objective of the proposed research is to isolate the mutated gene at the TG.KD transgene insertion locus and to understand the role of the isolated gene in the development of ovarian teratocarcinomas. Three specific aims are proposed: 1. To test the hypothesis that ovarian teratocarcinomas arise in the TG.KD line as the result of the transgene insertional inactivation or activation of a host gene, transcripts at the transgene integration locus will be identified, cloned, and characterized. 2. To assess when during germ cell development the mutant gene acts, F1 mice will be produced by crossing the TG.KD-bearing FVB/N inbred line to other inbred lines. The resulting teratocarcinomas will be genotyped using heteromorphic markers for the pericentromeric and distal regions of several chromosomes. The distribution of markers relative to that in F1 somatic cells will be used to determine the latest stage of meiosis reached during the formation of the teratocarcinomas. 3. To examine the phenotype of mice homozygous for the TG.KD transgene insertion. While hemizygous TG.KD transgenic mice have a propensity to develop ovarian teratocarcinomas, homozygous mice exhibit a perinatal lethality. Based on the working hypothesis that the same gene mutation underlies both the teratocarcinomas and the perinatal lethality, the phenotype of the homozygous embryos will be examined histologically to provide information regarding the function of the mutated gene.