Abstract
In situ hybridization is defined as one of the most useful and powerful methods to know where genes (e.g., mRNA, ncRNA) of interest are expressed in tissues. Expression of mRNA can be detected as blue or dark purple signals though hybridization, immunoreaction and coloring steps. Genome-wide approaches in various model animals have been conducted thoroughly, and have led to new research areas aimed at uncovering novel gene functions in cell differentiation and development. To elucidate gene function, spatiotemporal gene expression analysis is very important. Here I describe protocols of whole-mount and section in situ hybridization, and emphasize the relevance of optimizing temperature, and sodium concentration, in hybridization buffer and substrate to improve signal.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Noji S, Yamaai T, Koyama E, Nohno T, Fujimoto W, Arata J, Taniguchi S (1989) Expression of retinoic acid receptor genes in keratinizing front of skin. FEBS Lett 259:86–90
Riddle RD, Johnson RL, Laufer E, Tabin C (1993) Sonic hedgehog mediates the polarizing activity of the ZPA. Cell 75:1401–1416
Yokouchi Y, Sasaki H, Kuroiwa A (1991) Homeobox gene expression correlated with the bifurcation process of limb cartilage development. Nature 353:443–445
Bruneau BG, Bao ZZ, Tanaka M, Schott JJ, Izumo S, Cepko CL, Seidman JG, Seidman CE (2000) Cardiac expression of the ventricle-specific homeobox gene Irx4 is modulated by Nkx2-5 and dHand. Dev Biol 217:266–277
Acknowledgments
I thank F. Nakayama and D. Matsumura for technical assistance, and J. K. Takehchi for critical reading of the manuscript and useful advice.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Koshiba-Takeuchi, K. (2018). Whole-Mount and Section In Situ Hybridization in Mouse Embryos for Detecting mRNA Expression and Localization. In: Delgado-Olguin, P. (eds) Mouse Embryogenesis. Methods in Molecular Biology, vol 1752. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7714-7_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7714-7_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7713-0
Online ISBN: 978-1-4939-7714-7
eBook Packages: Springer Protocols