Chapter 16
Whole Mount in Situ Hybridization
for mRNA Detection in Chick Embryos
HIDEYO OHUCHI, AND SUMIHARE NOJI
Introduction
Analysis of the spatial expression patterns of genes in animal
embryos is essential to understanding functions of the genes
especially for developmental biology. In situ hybridization
with whole embryos is a powerful method to observe overall expression patterns of genes in embryos. A method for the whole
mount in situ hybridization was developed after non-radioactive
probes, especially digoxigenin-labeled RNA probes, became
commercially available. Furthermore, using two different
non-radioactive probes, expression patterns of two different
genes can be observed simultaneously in an embryo, as a pattern
of two colors. Sections of the stained embryos can be used to
identify cells expressing the mRNA in the embryo. Thus, this
method allows the identification of mRNA within intact embryos
at both regional and cellular levels.
Outline
The entire protocol of a whole-mount in situ hybridization is
illustrated in Fig. 1 (Noji et al. 1997).
Hideyo Ohuchi, The University of Tokushima, Department of Biological
Science and Technology, Faculty of Engineering, 2-1 Minami-Jyosanjima-cho, Tokushima City, 770, Japan; Correspondence to Sumihare Noji,
The University of Tokushima, Department of Biological Science and Technology, Faculty of Engineering, 2-1 Minami-Jyosanjima-cho, Tokushima
City, 770, Japan (phone +81-886-56-7528;fax +81-886-56-9074; e-mail noji@bio.tokushima-u.ac.jp)