Cardona Albert, Fernández Juani, Solana Jordi, Romero Rafael
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, 645 Diagonal, 08028, Barcelona, Spain.
Dev Genes Evol. 2005 Sep;215(9):482-88. doi: 10.1007/s00427-005-0003-1. Epub 2005 Jul 12.
The monitoring of gene expression is fundamental for understanding developmental biology. Here we report a successful experimental protocol for in situ hybridization in both whole-mount and sectioned planarian embryos. Conventional in situ hybridization techniques in developmental biology are used on whole-mount preparations. However, given that the inherent lack of external morphological markers in planarian embryos hinders the proper interpretation of gene expression data in whole-mount preparations, here we used sectioned material. We discuss the advantages of sectioned versus whole-mount preparations, namely, better probe penetration, improved tissue preservation, and the possibility to interpret gene expression in relation to internal morphological markers such as the epidermis, the embryonic and definitive pharynges, and the gastrodermis. Optimal fixatives and embedding methods for sectioning are also discussed.
基因表达监测对于理解发育生物学至关重要。在此，我们报告了一种在整体和切片涡虫胚胎中进行原位杂交的成功实验方案。发育生物学中的传统原位杂交技术用于整体标本。然而，鉴于涡虫胚胎中固有的缺乏外部形态标记物，这阻碍了对整体标本中基因表达数据的正确解读，因此我们在此使用了切片材料。我们讨论了切片标本与整体标本相比的优势，即更好的探针穿透性、改善的组织保存，以及根据内部形态标记物（如表皮、胚胎期和成虫期咽以及肠上皮）来解读基因表达的可能性。还讨论了用于切片的最佳固定剂和包埋方法。
