MUTANT LINES OF ZEBRAFISH HIGHLY SENSITIVE TO NEOPLASIA

对肿瘤高度敏感的斑马鱼突变系

• 批准号: 6677858
• 负责人: JAN M SPITSBERGEN
• 金额: $ 14.15万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6677858
• 关键词: DNA damage; N methyl N' nitro N nitrosoguanidine; alternatives to animals in research; bioassay; biological models; cancer risk; carcinogen testing; chemical carcinogen; chemical carcinogenesis; chemical related neoplasm /cancer; dimethylbenzanthracene; disease /disorder onset; environment related neoplasm /cancer; gene mutation; genetic susceptibility; immunocytochemistry; model design /development; morphometry; mutagen testing; mutant; neoplasm /cancer; nitrosourea; tumor suppressor genes; zebrafish

DESCRIPTION (provided by applicant): For decades, spontaneous and induced mutant animals have yielded valuable insights into the roles of specific genes in development and disease. Recently, transgenic and knockout mice, together with large-scale mutagenesis screens of model organisms such as the fruitfly, Drosophila, and the nematode, C. elegans, have contributed detailed insights into the roles of specific genes in development. These intensive studies reveal a remarkable degree of conservation of genetic strategies for control of cell differentiation, cell proliferation, and cell death, as well as pattern formation during development throughout all animals. Advantages of zebrafish for study of fundamental vertebrate genetic principles and molecular developmental genetics include small adult size, relative hardiness in small aquaria, high fecundity, external fertilization and development with accessibility of all developmental stages, highly transparent eggshells, rapid embryonic development, short generation times, ease of embryonal cloning to allow gynogenesis or androgenesis, and applicability for large-scale, rapid mutant screens. Intense worldwide research in zebrafish molecular development, genomics, and mutagenesis during the past decade has complemented the previous strong database on biology, physiology, and development of zebrafish creating a powerful system in which to answer fundamental questions in biology and medicine. The investigators will develop lines of zebrafish highly sensitive to specific histological types of neoplasia occurring in specific tissues. To develop these lines, the investigators will evaluate carcinogen-induced neoplasia in lines of zebrafish with inactivating mutations in orthologs of human tumor suppressor genes. The investigators will screen offspring of fish mutagenized with N-ethyl-N-nitrosourea (ENU), using temperature mediated heteroduplex analysis (TMHA) and the mismatch cleavage enzyme CELI to detect point mutations in zebrafish orthologs of the human tumor suppressor genes P53, RB1, WT1, PTEN, or NF1. The investigators will treat zebrafish embryos or fry of these mutant lines by immersion exposure to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or 7,12-dimethylbenz[a]anthracene (DMBA), then will evaluate neoplasm incidences six and 12 months later. These fish will provide a rapid and relatively inexpensive whole-animal bioassay system to detect carcinogenic risks to man from new materials as well as from environmental contaminants. These lines of zebrafish will provide an efficient and inexpensive system for development of new treatment and prevention strategies for cancer, and will help identify new genes controlling growth and differentiation.

描述（由申请人提供）： 几十年来，自发和诱发的突变动物已经对特定基因在发育和疾病中的作用产生了宝贵的见解。 最近，转基因和基因敲除小鼠以及大规模的诱变筛选（例如果蝇，果蝇和线虫秀丽隐杆线虫）的大型诱变筛选，对特定基因在发育中的作用有了详细的见解。 这些密集研究揭示了对控制细胞分化，细胞增殖和细胞死亡的控制的显着保存，以及所有动物发育过程中的模式形成。 Advantages of zebrafish for study of fundamental vertebrate genetic principles and molecular developmental genetics include small adult size, relative hardiness in small aquaria, high fecundity, external fertilization and development with accessibility of all developmental stages, highly transparent eggshells, rapid embryonic development, short generation times, ease of embryonal cloning to allow gynogenesis or androgenesis, and applicability for大规模快速突变屏幕。 在过去的十年中，全球遍布斑马鱼分子发育，基因组学和诱变的全球研究已经补充了先前关于生物学，生理学和斑马鱼发展的强大数据库，创造了一个有力的系统，可以回答生物学和医学的基本问题。 研究人员将开发斑马鱼对特定组织中发生的肿瘤的特定组织学类型高度敏感的线条。 为了发展这些线路，研究人员将评估斑马鱼线中致癌物诱导的肿瘤，并在人类肿瘤抑制基因的直系同源物中灭活突变。 研究人员将使用温度介导的异质动力分析（TMHA）和不匹配的裂解酶CELI来检测人类肿瘤抑制基因斑马群正径中的点突变，以筛选用N-乙基N-硝基库（ENU）的鱼类的后代，并使用温度介导的杂化分析（TMHA）和不匹配的裂解酶突变来抑制点突变。 研究人员将通过浸入到N-甲基N'N'N硝基-N-硝基苯胺（MNNG）或7,12-二甲基苯甲酸盐[A]蒽（DMBA）（DMBA）中来治疗这些突变系的斑马鱼胚胎或油炸，然后评估六个月和12个月后的肿瘤性率。 这些鱼将提供快速且相对便宜的全动物生物测定系统，以发现新材料以及环境污染物的人的致癌风险。 这些斑马鱼的线条将为开发新的治疗和预防癌症的策略提供高效且廉价的系统，并有助于确定控制生长和分化的新基因。