大性地中海星虫的整体原位杂交

Whole-Mount In Situ Hybridization in Large Sexual Schmidtea mediterranea.

机构信息

Department of Tissue Dynamics and Regeneration, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany.

European Molecular Biology Laboratory, Heidelberg, Germany.

出版信息

Methods Mol Biol. 2023;2680:107-119. doi: 10.1007/978-1-0716-3275-8_7.

DOI:10.1007/978-1-0716-3275-8_7

PMID:37428374

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）可用于可视化内源性 RNA。对于扁形动物，模式物种地中海扁形虫和日本三角涡虫的小型动物（>5 毫米）有可靠的 WISH 方案。然而，用于生殖系发育和功能研究的地中海扁形虫的性系个体的体型要大得多，超过 2 厘米。现有的整体 WISH 方案对于如此大的标本并不理想，因为组织通透性不足。在这里，我们描述了一种针对 12-16 毫米长的性成熟地中海扁形虫个体的稳健 WISH 方案，该方案可以作为将 WISH 适应于其他大型扁形动物物种的起点。