Abstract
Over the last several decades, an understanding of the events involved in embryonic pattern formation has been extended from an anatomic to a molecular level. In particular, the ability to examine the spatial and temporal expression of specific genes has aided greatly the analysis of their function. One of the most powerful techniques for determining gene-expression patterns is in situ hybridization, a method that relies on the hybridization of labeled antisense RNA probes to detect endogenous RNA transcripts at their sites of expression (1).
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Notes
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1
We have tried to indicate the source of most of the critical reagents that we have used. Similar products are often available from other sources and may work equally well.
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2
We have found that the type of slide is critical for maintaining intact sections through the many washes required by this protocol, especially for small tissue sections (e.g., E10.5 and earlier).
References
Wilkinson, D. G. (1992) In Situ Hybridization: A Practical Approach. IRL, Oxford, UK.
Li, H.-S., Yang, J.-M., Jacobson, R. D., Pasko, D., and Sundin, O. (1994) Pax-6 is first expressed in a region of ectoderm anterior to the early neural plate: implications for stepwise determination of the lens. Dev. Biol. 162, 181–194.
Rosen, B. and Beddington, R. S. P. (1993) Whole mount in situ hybridization in the mouse embryo: gene expression in three dimensions. Trends Genet. 9, 162–168.
Epstein, J. A., Shapiro, D. N., Cheng, J., Lam, P. Y., and Maas, R. L. (1996) Pax3 modulates expression of the c-Met receptor during limb muscle development. Proc. Natl. Acad. Sci. USA 93, 4213–4218.
Xu, P. X., Cheng, J., Epstein, J. A., and Maas, R. L. (1997) Mouse Eya genes are expressed during limb tendon development and encode a transcriptional activation function. Proc. Natl. Acad. Sci. USA 94, 11,974–11,979.
Lutz, B., Kuratani, S., Cooney, A. J., Wawersik, S., Tsai, S. Y., Eichele, G., and Tsai, M. J. (1994) Developmental regulation of the orphan receptor COUP-TFII gene in spinal motor neurons. Development 120, 25–36.
Kuratani, S., Martin, J. F., Wawersik, S., Lilly, B., Eichele, G., and Olson, E. N. (1994) The expression pattern of the chick homeobox gene gMHox suggests a role in patterning of the limbs and face and in compartmentalization of somites. Dev. Biol. 161, 357–369.
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© 2000 Humana Press Inc., Totowa, NJ
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Wawersik, S., Epstein, J.A. (2000). Gene Expression Analysis by In Situ Hybridization. In: Tuan, R.S., Lo, C.W. (eds) Developmental Biology Protocols. Methods in Molecular Biology™, vol 137. Humana Press. https://doi.org/10.1385/1-59259-066-7:87
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DOI: https://doi.org/10.1385/1-59259-066-7:87
Publisher Name: Humana Press
Print ISBN: 978-0-89603-854-7
Online ISBN: 978-1-59259-066-7
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