An in situ hybridization protocol for planarian embryos:: monitoring myosin heavy chain gene expression

被引:29
作者
Cardona, A [1 ]
Fernández, J [1 ]
Solana, J [1 ]
Romero, R [1 ]
机构
[1] Univ Barcelona, Fac Biol, Dept Genet, E-08028 Barcelona, Spain
关键词
planaria; myosin; paraffin section; in situ hybridization; flatworm;
D O I
10.1007/s00427-005-0003-1
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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.
引用
收藏
页码:482 / 488
页数:7
相关论文
共 18 条
[1]   Structure of the planarian central nervous system (CNS) revealed by neuronal cell markers [J].
Agata, K ;
Soejima, Y ;
Kato, K ;
Kobayashi, C ;
Umesono, Y ;
Watanabe, K .
ZOOLOGICAL SCIENCE, 1998, 15 (03) :433-440
[2]  
ALVARADO AS, 2003, PROPOSAL SEQUENCING
[3]  
BAGUNA J, 1989, DEVELOPMENT, V107, P77
[4]  
Benazzi M., 1982, P151
[5]   Nonradioactive in situ hybridization to Xenopus tissue sections [J].
Butler, K ;
Zorn, AM ;
Gurdon, JB .
METHODS, 2001, 23 (04) :303-312
[6]   The embryonic development of the triclad Schmidtea polychroa [J].
Cardona, A ;
Hartenstein, V ;
Romero, R .
DEVELOPMENT GENES AND EVOLUTION, 2005, 215 (03) :109-131
[7]   Intercalary muscle cell renewal in planarian pharynx [J].
Cebrià, F ;
Bueno, D ;
Reigada, S ;
Romero, R .
DEVELOPMENT GENES AND EVOLUTION, 1999, 209 (04) :249-253
[8]   Myocyte differentiation and body wall muscle regeneration in the planarian Girardia tigrina [J].
Cebria, F ;
Vispo, M ;
Newmark, P ;
Bueno, D ;
Romero, R .
DEVELOPMENT GENES AND EVOLUTION, 1997, 207 (05) :306-316
[9]  
CEBRIA F, 2000, THESIS U BARCELONA
[10]   The embryonic development of the rhabdocoel flatworm Mesostoma lingua (Abildgaard, 1789) [J].
Hartenstein, V ;
Ehlers, U .
DEVELOPMENT GENES AND EVOLUTION, 2000, 210 (8-9) :399-415