Novel Use of Zebrafish for Apoptosis Assay

斑马鱼在细胞凋亡检测中的新用途

• 批准号: 6587090
• 负责人: Chuenlei Parng
• 金额: $ 51.49万
• 依托单位: PHYLONIX PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-15 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6587090
• 关键词: acridines; apoptosis; bioassay; biological models; biomedical automation; cell differentiation; cysteine endopeptidases; drug screening /evaluation; embryo /fetus tissue /cell culture; flow cytometry; fluorescent dye /probe; high throughput technology; technology /technique development; zebrafish

DESCRIPTION (provided by applicant): Inappropriate apoptosis is the cause of many human diseases, including cancer, neuronal degeneration, hearing loss, organ failure, myelodysplastic syndrome, multiple sclerosis, type 1 diabetes mellitus, thyroiditis, and ulcerative colitis. Compounds that affect apoptosis are potential therapeutics. Cell based assays are widely used in drug development, however, many promising candidates fail in subsequent mammalian testing. Zebrafish assays can serve as an intermediate step between cellular evaluation and mammalian testing. Zebrafish has several advantages, including transparent embryos for easy visual analysis, rapid embryonic development and low cost. Furthermore, similar pharmacological responses to those in the mammalian apoptotic machinery are present in zebrafish. Currently the standard whole animal apoptosis assay is TUNEL, a histochemical staining technique which detects apoptotic cells in sectioned tissue. This assay is time-consuming and permits examination at only a single time point. Since many degenerative diseases occur in stages, assay formats that permit continuous examination of changes in the apoptosis pattern and the duration of drug effects are needed. Using zebrafish embryos, this proposal will develop a quantitative microplate assay for high throughput screening of compounds. Complementary visual and flow cytometry based assays will also be developed to provide comprehensive analysis of apoptosis in zebrafish.

描述（由申请人提供）： 不适当的凋亡是许多人类疾病的原因，包括癌症，神经元变性，听力损失，器官衰竭，骨髓增生症状综合征，多发性硬化症，1型糖尿病，甲状腺胶质炎，甲状腺炎和溃疡性结肠炎。影响凋亡的化合物是潜在的治疗剂。基于细胞的测定在药物开发中广泛使用，但是，许多有前途的候选者在随后的哺乳动物测试中失败。斑马鱼测定可以作为细胞评估和哺乳动物测试之间的中间步骤。斑马鱼具有多个优点，包括透明的胚胎，可轻松视觉分析，快速胚胎发育和低成本。此外，斑马鱼中存在对哺乳动物凋亡机制中的药理学反应。目前，整个动物凋亡测定法是TUNEL，TUNEL是一种组织化学染色技术，可检测到截面组织中的凋亡细胞。该测定时间是耗时的，并且只能在一个时间点进行检查。由于许多退化性疾病分阶段发生，因此需要连续检查凋亡模式的变化和药物效应持续时间的测定格式。使用斑马鱼胚胎，该建议将开发一种定量的微镀码分析，用于高吞吐物的化合物。还将开发互补的视觉和流式细胞仪测定法，以提供斑马鱼凋亡的全面分析。