Genetic Control of Skeletogenesis in the Zebrafish

描述：(申请人摘要)人类骨骼发育不良，尽管 重要的致病原因，也被证明是一个丰富的信息来源 关于骨骼发育和功能的生物学。突变的基因 编码广泛的蛋白质，包括转录因子、信号 分子和结构蛋白。然而，负责许多疾病的基因 影响骨骼的疾病仍未确定。找出类似的 一个易驯服的模型生物体中的突变不仅可以进行详细的研究 深入研究潜在的疾病机制，但也可能有助于克隆 人类疾病基因。最近的研究证明斑马鱼Danio Rerio 是进行大规模突变的最实用的脊椎动物 屏幕。然而，以前的筛查都集中在识别突变上 影响早期发育的。我们建议筛选影响基因的突变 成年斑马鱼骨骼的结构和形态，用X射线照相法 一种检查活体动物骨骼的有效方法。我们还建议 利用容易看到和容易接近的斑马鱼胚胎，进行筛选 专门影响成骨细胞的突变。我们将做一些线条 表达绿色荧光蛋白(GFP)的转基因鱼 转录因子cbfal的启动子，在早期成骨细胞中表达。vbl.使用 以这条线为背景，我们将筛选影响图案的突变 在成骨细胞可以被区分之前的阶段，绿色荧光蛋白的表达 从形态上看。使用这两种筛选方法，我们的目标是识别基因 控制骨骼形成的所有阶段，从构图到形态形成。 此外，我们正专注于识别曾被 不太可能在以前的屏幕中被隔离，因此进一步扩大了 斑马鱼作为模式遗传系统的有用性。通过以下方式获得的知识 这项工作应该有助于鉴定导致骨骼的人类突变 异常，也有助于更好地理解常见问题 影响骨骼，如骨质疏松症、关节炎和再生后 受伤。

DESCRIPTION: (Applicant's Abstract) Human skeletal dysplasias, although a significant cause of morbidity, have also proven a rich source of information about the biology of skeletal development and function. The mutated genes encode a wide range of proteins, including transcription factors, signaling molecules, and structural proteins. However, the genes responsible for many diseases affecting the skeleton remain unidentified. To identify similar mutations in a tractable model organism would not only allow detailed study into the underlying disease mechanisms, but could also aid in the cloning of human disease genes. Recent studies have proven the zebrafish, Danio rerio, to be the most practical vertebrate organism for performing large-scale mutational screens. However, previous screens have focused on identifying mutations affecting early development. We propose to screen for mutations affecting the structure and morphology of the adult zebrafish skeleton, using radiography as an efficient method to examine the skeleton in live animals. We also propose to take advantage of the easily visible and accessible zebrafish embryo, to screen for mutations specifically affecting osteoblasts. We will make lines of transgenic fish expressing green fluorescent protein (GFP) under control of the promoter of cbfal, a transcription factor expressed in early osteoblasts. Using this line as a background, we will screen for mutations affecting the pattern of GFP expression, at stages before the osteoblasts can be distinguished morphologically. Using these two screening approaches, we aim to identify genes controlling all stages of skeletogenesis, from patterning to morphogenesis. Additionally, we are focusing on identifying classes of mutants that were unlikely to have been isolated in previous screens, thus further expanding the usefulness of the zebrafish as a model genetic system. The knowledge gained by this work should aid in the identification of human mutations causing skeletal abnormalities, and also lead to better understanding of common problems affecting the skeleton, such as osteoporosis, arthritis, and regeneration after injury.