An in situ hybridization protocol for planarian embryos: monitoring myosin heavy chain gene expression.
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.
Cardona
Albert
A
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, 645 Diagonal, 08028, Barcelona, Spain. acardoto7@bio.ub.edu
Fernández
Juani
J
Solana
Jordi
J
Romero
Rafael
R
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000201:enzymology; D004817:Epidermis / Q000201:enzymology; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization; D018995:Myosin Heavy Chains / Q000235:genetics* / Q000378:metabolism; D010932:Planarians / Q000196:embryology* / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods
D018995:Myosin Heavy Chains