Enhanced Image Analysis of a Transparent Fish Model

描述(由申请者提供)：越来越多的人呼吁使用替代动物模型作为评估人类健康风险的替代品。透明青衣是一种独特的动物模型，我们认为它在评估此类风险方面具有巨大的潜力。这种动物模型在胚胎、幼年和成年生命阶段是透明的，活体动物的主要内脏通过体壁可见。有了透明青竹，目前在胚胎发育过程中现有替代模型中可用的表型分析的简便性现在可能扩展到更晚的生命阶段。分子、细胞、组织和器官/系统水平的结构和功能变化可以非侵入性地成像，从而在整个生命过程中极大地增强表型特征。提出了四个具体目标：1)明确青山肝脏的生长发育，强调肝小管与微血管的关系。2)肝小管内胆汁形成过程中肝细胞与胆管上皮细胞相互作用的图像。3)构建细胞类型特异的透明青色转基因细胞，以扩大对肝脏发育/重塑和生长的非侵入性检测。4)通过暴露于已知的以肝内胆汁功能为靶标的参比肝毒素，以及在单独的个体中，暴露于可导致青竹肝脏变性和肿瘤性损害的已知致癌物质，在透明青竹幼虫中产生经证实的肝脏损伤。青竹压缩的生命周期，再加上其透明的特征，使这一模型特别适合于研究胚胎后发育、衰老、毒性、突变和致癌。尤其重要的是一种评估早期生命阶段暴露的表现的模型，在该模型中，最初的分子紊乱可以与成人功能障碍分开很长一段时间。拟议研究中的努力将集中在扩展对肝脏发育/重塑和生长的非侵入性检测过程。我们之所以选择研究肝脏，是因为我们对这一器官系统有广泛的了解，而且它在以下方面的重要性：营养物质和其他内源性分子的吸收、代谢、储存和再分配，外源物质的代谢，以及胆汁的形成和排泄。因此，在这一应用中，我们将建立特定的分子成像技术来研究器官系统的生长和发育，并增强该模型在研究人类疾病影响方面的潜力。

DESCRIPTION (provided by applicant): There is an increasing call for alternative animal models as surrogates for assessing risks to human health. The see-through medaka is a unique animal model, which we believe has enormous potential for assessing such risks. This animal model is transparent in embryonic, juvenile and adult life stages with principal internal organs visible through the body wall in the living animals. With the see-through medaka, ease of phenotypic analysis currently available in existing alternative models during embryogenesis may now be extended to later life stages. Structural and functional changes at molecular, cellular, tissue and organ/system levels may be imaged non-invasively leading to greatly enhanced phenotypic characterization throughout life. Four specific aims are proposed: 1) Define medaka liver growth and development emphasizing relationship of hepatic tubules to microvasculature. 2) Image hepatocyte and biliary epithelial cell interaction in bile formation within the hepatic tubule. 3) Construct cell type specific transgenic see-through medaka to extend noninvasive detection of liver development/remodeling and growth. 4) To produce verified hepatic injury in larval see-through medaka by exposure to a reference hepatotoxin known to target intrahepatic biliary function, and, in separate individuals, to a known carcinogen that results in degenerative and neoplastic lesions in medaka liver. The compressed life cycle of the medaka, when coupled with its transparent features, makes this model particularly well suited to study post-embryonic development, aging, toxicity, mutagenesis and carcinogenesis. Particularly important is a model for evaluating manifestations of early life stage exposure where initial molecular derangement may be separated from adult dysfunction by long intervals. Effort in the proposed research will focus on processes to extend noninvasive detection of liver development/remodeling and growth. We have chosen to study the liver because of our extensive knowledge with this organ system and it's importance in: uptake, metabolism, storage and redistribution of nutrients and other endogenous molecules, metabolism of xenobiotics, and formation and excretion of bile. Thus, in this application, we will establish specific molecular imaging technologies to study organ system growth and development and to enhance this model's potential to study effects of human disease.