COREGULATION OF ALK8 AND BMP4 IN ZEBRAFISH TOOTH FORMATI

斑马鱼牙齿形态中 ALK8 和 BMP4 的共调控

• 批准号: 6175801
• 负责人: TRACIE L FERREIRA
• 金额: $ 4.09万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6175801
• 关键词: DNA footprinting; bone morphogenetic proteins; dental development; developmental genetics; gel mobility shift assay; gene expression; genetic promoter element; genetic regulation; genetic regulatory element; green fluorescent proteins; growth factor receptors; immunocytochemistry; in situ hybridization; microinjections; polymerase chain reaction; protein kinase; protein structure function; reporter genes; transforming growth factors; zebrafish

Although the stages of zebrafish development have been well documented, the molecular mechanisms underlying the cellular movements, proliferation and differentiation are not well defined. The zALK-8 (zebrafish Activin Receptor-Like Kinase) gene predicts a protein with a serine/threonine kinase domain similar to other TGF-beta superfamily receptors. Initial characterization of partial genomic clones revealed a conserved intronic sequence between zALK-8 and the zebrafish BMP-4 (Bone Morphogenic Protein) gene. Preliminary immunohistochemical analysis indicates that zALK-8 is developmentally regulated and is coordinately expressed with zBMP-4 during tooth development, positioning it well to play a role in tooth development. We hypothesize that the coordinate expression of zALK-8 and zBMP-4 is essential for normal tooth development. We propose the following specific aims to test this hypothesis. First, we will fully characterize the promoter and intronic sequences of zALK-8 gene to facilitate identification of regulatory domains. Promoter sequences and other conserved domains will then be examined in reporter constructs to identify potential enhancer regions and binding domains for cis-acting elements. The coordinate expression and regulation of the zALk-8 and zBMP-4 genes in tooth development will be examined using reporter constructs of GFP for zALk-8 and YFP for zBMP-4 allowing easy visualization of overlapping and distinct domains of expression.

尽管斑马鱼发育的阶段已经充分证明，但细胞运动的分子机制，增殖和分化尚未得到很好的定义。 Zalk-8（斑马鱼激活素受体样激酶）基因预测具有丝氨酸/苏氨酸激酶结构域的蛋白质，类似于其他TGF-β-beta超家族受体。 部分基因组克隆的最初表征揭示了Zalk-8和斑马鱼BMP-4（骨形态发生蛋白）基因之间的保守内在序列。 初步免疫组织化学分析表明，Zalk-8在发育过程中受到发育调节，并在牙齿发育过程中与ZBMP-4协调表达，可以很好地定位在牙齿发育中发挥作用。 我们假设Zalk-8和ZBMP-4的坐标表达对于正常牙齿发育至关重要。 我们提出以下特定旨在检验该假设的特定旨在。 首先，我们将充分表征Zalk-8基因的启动子和内含子序列，以促进调节域的鉴定。然后，将在报告基因构建体中检查启动子序列和其他保守域，以确定潜在的增强子区域和顺式作用元件的结合域。 Zalk-8和ZBMP-4基因在牙齿发育中的坐标表达和调节将使用ZALK-8和ZBMP-4的YFP的记者构建器进行检查，以便于轻松地可视化重叠和不同表达域。