Abstract
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.
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Acknowledgements
The authors thank Francesc Cebrià for his previous work on planarian muscle development, which produced the gene fragments from which probes were generated for this study. A. Cardona thanks Volker Hartenstein for his comments and assistance in preparing this manuscript and the anonymous reviewers for their suggestions. This work was supported by grant BFU2004-05015 from the PNIC-DIT 2004-07 by the Ministerio de Educación y Ciencia to R. Romero.
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Communicated by V. Hartenstein
A. Cardona and J. Fernández have contributed equally to this work.
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Cardona, A., Fernández, J., Solana, J. et al. An in situ hybridization protocol for planarian embryos: monitoring myosin heavy chain gene expression. Dev Genes Evol 215, 482–488 (2005). https://doi.org/10.1007/s00427-005-0003-1
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DOI: https://doi.org/10.1007/s00427-005-0003-1