"Xenopus Microarrays: A Tool for Molecular Toxicology"

描述（由申请人提供）：在美国，每年大约有12万婴儿出生时患有重大出生缺陷，遗传和环境因素起着重要作用。胚胎分化途径在脊椎动物中是高度保守的。非洲爪蟾胚胎是一种易于接触化学物质的成熟毒理学模型。实验室间的研究证明了青蛙胚胎畸形试验-爪蟾（FETAX）的可靠性。非洲爪蟾微阵列分析作为药物筛选和环境毒理学机制研究的多功能工具具有很大的前景。我们的长期目标是开发爪蟾微阵列，根据基因表达谱对环境毒物进行分类，并发现环境污染物的标记。从NIEHS获得约18500个EST克隆。第一阶段完成的大约1200个克隆的初步研究表明，使用爪蟾微阵列识别早期胚胎发育中对phorbol 12-肉豆蔻酸13-醋酸酯（PMA）敏感的基因是可行的。在第二阶段，我们将开发高密度cDNA微阵列，并鉴定受7类有毒物质调控的基因。这些结果将用于产生诊断子阵列。我们还将生产种间外推芯片，研究毒物对基因调控的保护作用。此外，还将探讨利用热带野蓟芯片进行热带野蓟遗传操作研究的可行性。

DESCRIPTION (provided by applicant): Each year, approximately 120,000 babies are born in the US with a major birth defect and genetic and environmental factors play an important role. Embryonic differentiation pathways are highly conserved among vertebrates. The Xenopus laevis embryo is an established toxicology model with ease of chemical exposure. Inter-laboratory studies demonstrated the reliability of the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Xenopus microarray analysis holds great promise as a versatile tool for drug screening and mechanistic studies of environmental toxicology. Our long-term goal is to develop Xenopus microarrays, categorize environmental toxicants according to their gene expression profiles and discover markers of environmental contaminants. About 18,500 EST clones were obtained from NIEHS. Initial studies completed in the Phase I with about1,200 clones demonstrate the feasibility of using Xenopus microarrays to identify genes responsive to phorbol 12-myristate 13-acetate (PMA) in early embryogenesis. In the Phase II, we will develop high-density cDNA microarrays and identify genes regulated by 7 classes of toxicants. These results will be utilized to produce diagnostic subarrays. We will also produce interspecies extrapolation chips to study conservation of gene regulation by toxicants. In addition, the feasibility of use of X. laevis chips for a X. tropicalis genetic manipulation study will be explored.