An in situ hybridization protocol for planarian embryos:: monitoring myosin heavy chain gene expression
An in situ hybridization protocol for planarian embryos:: monitoring myosin heavy chain gene expression
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DOI：
10.1007/s00427-005-0003-1
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发表时间：
2005-09-01
影响因子：
2.4
通讯作者：
Romero, R
中科院分区：
文献类型：
--
作者：
Cardona, A;Fernández, J;Romero, R
基因表达的监测是理解发育生物学的基础。在这里，我们报告了一个成功的实验方案原位杂交在整体安装和切片的涡虫胚胎。常规的原位杂交技术在发育生物学中用于整装制备。然而，鉴于在真涡虫胚胎中固有的外部形态标记的缺乏阻碍了在整体安装准备中基因表达数据的正确解释，在这里我们使用切片材料。我们讨论的优势，切片与整装制剂，即更好的探头渗透，改善组织保存，并解释基因表达的可能性，内部形态标志物，如表皮，胚胎和明确的咽，和gastrodermis。最佳固定剂和嵌入方法的切片进行了讨论。
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.