Enhanced Image Analysis of a Transparent Fish Model

描述（由申请方提供）：越来越多的人要求替代动物模型作为评估人类健康风险的替代物。透明青鳉是一种独特的动物模型，我们相信它在评估此类风险方面具有巨大的潜力。该动物模型在胚胎、幼年和成年生命阶段是透明的，通过活体动物的体壁可以看到主要的内脏器官。有了透明青鳉，目前在胚胎发生期间现有替代模型中可用的表型分析的便利性现在可以扩展到以后的生命阶段。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，从而大大增强整个生命过程中的表型表征。提出了四个具体目标：1）定义青鳉肝脏生长和发育，强调肝小管与微血管的关系。2)肝细胞和胆管上皮细胞在肝小管内胆汁形成中的相互作用。3)构建细胞类型特异性转基因透明青鳉以扩展肝脏发育/重塑和生长的无创检测。4)通过暴露于已知靶向肝内胆汁功能的参考肝毒素，以及在单独个体中暴露于导致青鳉肝脏退行性和肿瘤性病变的已知致癌物，在青鳉幼体透明青鳉中产生经验证的肝损伤。青鳉的压缩生命周期，再加上其透明的特点，使这个模型特别适合研究胚胎后发育，衰老，毒性，诱变和致癌作用。特别重要的是一个模型，用于评估早期生命阶段暴露的表现，其中最初的分子紊乱可能会分开成人功能障碍的长时间间隔。拟议研究的努力将集中在扩展肝脏发育/重塑和生长的非侵入性检测的过程上。我们选择研究肝脏是因为我们对这个器官系统有广泛的了解，它在以下方面很重要：营养物质和其他内源性分子的吸收、代谢、储存和再分配，外源性物质的代谢，以及胆汁的形成和排泄。因此，在本申请中，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，并增强该模型研究人类疾病影响的潜力。

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.