Zebrafish Based Model for Gastrointestinal Physiology

基于斑马鱼的胃肠生理学模型

• 批准号: 6953392
• 负责人: ADAM J RICH
• 金额: $ 21.53万
• 依托单位: COLLEGE AT BROCKPORT
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6953392
• 关键词: biological models; digital imaging; electron microscopy; gastrointestinal function; gastrointestinal motility /pressure; immunocytochemistry; intestinal villi; model design /development; physiology; zebrafish

DESCRIPTION (provided by applicant): The overall goal of this new AREA (R-15) research proposal is to develop, validate, and utilize a zebrafish-based model system to elucidate the cellular and molecular details of gastrointestinal (Gl) function with an emphasis on motility. A new model system exploring Gl physiology is needed because current therapies for Gl motility disorders are lacking. A zebrafish-based model system will provide several important experimental advantages, such as the direct visualization of Gl motility in optically transparent larvae and the opportunity to utilize a forward genetic approach to identify novel molecular components determining Gl motility. The Gl tract appears functional by 5 days post-fertilization when larvae swallow food, exhibit peristalsis, and absorb nutrients. In Specific Aim 1 the cellular components that regulate Gl motility in mammals, interstitial cells of Cajal (ICC), will be characterized in larvae and adults, using immunohistochemical procedures. Expression of the kit gene specifically identifies ICC, and kit expression will identify putative ICC in the zebrafish. Development of ICC, or pacemaker cells, is expected to coincide with development of spontaneous contractions, similar to other vertebrates. The anatomical position of ICC, intermediary between enteric neurons and Gl smooth muscles, indicates the functional role of integration and coordination of motility in humans. Anatomical features of putative ICC cells and networks will be determined. In Specific Aim 2 the functional significance of kit-positive cells will be determined using wild type and null kit mutant zebrafish. Normal motility patterns, including the frequency of spontaneous contractions and the time for food to clear the Gl tract, will be determined. The dependence of motility on the pacemaker cell will be examined using zebrafish with inactivating kit mutations, which lesion ICC in other vertebrate models. A zebrafish based model system has the potential for the identification of novel genes regulating Gl motility, which may benefit treatment for disorganized motility.

描述（由申请人提供）：这项新的AREA（R-15）研究提案的总体目标是开发、验证和利用基于斑马鱼的模型系统，以阐明胃肠道（GI）功能的细胞和分子细节，重点是运动。需要探索GI生理学的新模型系统，因为目前缺乏GI运动障碍的治疗。基于斑马鱼的模型系统将提供几个重要的实验优势，例如光学透明幼虫中GI运动的直接可视化和利用正向遗传方法来鉴定决定GI运动的新分子组分的机会。胃肠道在受精后5天出现功能，此时幼虫吞咽食物，表现出消化，并吸收营养。在具体目标1中，将使用免疫组织化学程序在幼虫和成虫中表征调节哺乳动物中GI运动的细胞组分，Cajal间质细胞（ICC）。kit基因的表达特异性地鉴定ICC，并且kit表达将鉴定斑马鱼中推定的ICC。ICC或起搏细胞的发育预计与自发收缩的发育一致，类似于其他脊椎动物。ICC的解剖学位置是肠神经元和GI平滑肌之间的中介，表明了人类运动的整合和协调的功能作用。将确定推定的ICC细胞和网络的解剖特征。在特定目标2中，将使用野生型和无效kit突变体斑马鱼确定kit阳性细胞的功能意义。将确定正常运动模式，包括自发收缩的频率和食物清除胃肠道的时间。将使用具有灭活试剂盒突变的斑马鱼来检查运动性对起搏细胞的依赖性，所述灭活试剂盒突变在其他脊椎动物模型中损害ICC。基于斑马鱼的模型系统具有鉴定调节GI运动的新基因的潜力，这可能有益于治疗紊乱的运动。