Simple blood-feeding method for live imaging of gut tube remodeling in regenerating planarians.
Simple blood-feeding method for live imaging of gut tube remodeling in regenerating planarians.
复制标题
用于再生涡虫肠管重塑实时成像的简单血液喂养方法。
DOI：
10.1111/dgd.12270
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Umesono Y.
中科院分区：
文献类型：
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作者：
Hosoda K;Morimoto M;Motoishi M;Nishimura O;Agata K;Umesono Y.
活细胞成像技术是研究动物体内发育和再生过程中细胞动力学的有效手段。然而，很少有现场成像方法已被报道用于研究真涡虫再生。在这里，我们开发了一种简单的方法，稳定的可视化再生过程中的一个活的淡水涡虫，三角洲日本肠管重塑。当涡虫被喂食血液几次时，在正常明场照明下，活的完整动物的肠道分支很好地可视化。有趣的是，来自这些彩色的涡虫的尾巴片段，使连续观察的过程中形成的一个单一的前肠分支在前咽区域从预先存在的两个后肠分支在同一个活的动物在头部再生。此外，我们将这种方法与RNA干扰（RNAi）相结合，从而表明我们鉴定的日本盘尾轮虫raf相关基因（DjrafA）和mek相关基因（DjmekA）在涡虫再生过程中细胞外信号调节激酶（ERK）信号的激活中起主要作用，正如在使用血液喂养而没有免疫组织化学检测的节省时间的初始筛选中，他们的RNAi诱导的肠管重塑缺陷所表明的那样，内脏因此，这种血液喂养方法可用于肠道重塑的实时成像，并为涡虫再生研究领域提供了一个进步。
Live cell imaging is a powerful technique to study cellular dynamicsin vivoduring animal development and regeneration. However, few live imaging methods have been reported for studying planarian regeneration. Here, we developed a simple method for steady visualization of gut tube remodeling during regeneration of a living freshwater planarian,Dugesia japonica. When planarians were fed blood several times, gut branches were well‐visualized in living intact animals under normal bright‐field illumination. Interestingly, tail fragments derived from these colored planarians enabled successive observation of the processes of the formation of a single anterior gut branch in the prepharyngeal region from the preexisting two posterior gut branches in the same living animals during head regeneration. Furthermore, we combined this method and RNA interference (RNAi) and thereby showed that aD. japonica raf‐related gene (DjrafA) andmek‐related gene (DjmekA) we identified both play a major role in the activation of extracellular signal‐regulated kinase (ERK) signaling during planarian regeneration, as indicated by their RNAi‐induced defects on gut tube remodeling in a time‐saving initial screening using blood‐feeding without immunohistochemical detection of the gut. Thus, this blood‐feeding method is useful for live imaging of gut tube remodeling, and provides an advance for the field of regeneration study in planarians.