Abstract:
Whole-mount in situ hybridization (WISH), colorimetric or fluorescent (FISH), allows for the visualization of endogenous RNA. For planarians, robust WISH protocols exist for small-sized animals (>5 mm) of the model species Schmidtea mediterranea and Dugesia japonica. However, the sexual strain of Schmidtea mediterranea studied for germline development and function reaches much larger body sizes in excess of 2 cm. The existing whole-mount WISH protocols are not optimal for such large specimens, owing to insufficient tissue permeabilization. Here, we describe a robust WISH protocol for 12-16 mm long sexually mature Schmidtea mediterranea individuals that could serve as a starting point for adapting WISH to other large planarian species.
摘要:
整体原位杂交(WISH),比色或荧光(FISH),允许内源性RNA的可视化。对于涡虫来说,对于模型物种Schmidteamediterranea和Dugesiajaponica的小型动物(>5mm),存在可靠的WISH方案。然而,对种系发育和功能进行研究的Schmidteamediterranea的性株达到更大的体型,超过2厘米。现有的整体安装WISH协议对于如此大的标本并不是最佳的,由于组织渗透性不足。这里,我们描述了一种针对12-16毫米长的性成熟Schmidteamediterranea个体的强大WISH协议,该协议可以作为使WISH适应其他大型涡虫物种的起点。