Abstract
The skin as a surface organ is uniquely accessible for whole embryo/fetal analyzes of developmental changes, such as gene induction, protein expression, formation of epidermal-derived appendages such as hair follicles, and formation of the protective barrier. Such analyses have emphasized the heterogenous nature of skin development, perhaps not surprising because epidermal development is programmed by heterogenous underlying mesenchyme. It is necessary to account for this heterogeneity by precisely matching body sites when correlating sequential events during development, for example, the activation of gene expression. In this chapter, protocols designed to assay whole-mount in situ hybridization and whole-mount barrier formation are presented. Formation of the protective barrier is the endpoint of epidermal terminal differentiation and defects in this process are reflected in failure, acceleration, or delay in barrier formation. Hence, these latter assays are of particular value as a rapid initial assay for epidermal developmental defects in genetically modified organisms.
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Hardman, M. J., Sisi, P., Banbury, D. N., and Byrne, C. (1998) Patterned acquisition of skin barrier function during development. Development 125, 1541–1552.
Xu, Q., and Wilkinson, D. G. (1997) In situ hybridization of mRNA with hapten labelled probes, in In Situ Hybridization, A Practical Approach, 2nd ed. (Wilkinson, D. G., ed.), IRL Press, Oxford, pp. 87–106.
Conlon, R. A. and Rossant, J. (1992) Exogenous retinoic acid rapidly induces anterior ectopic expression of murine Hox-2 genes in vivo. Development 116, 357–368.
Jowett, T. (1997) Two colour in situ hybridisation, in In Situ Hybridization, A Practical Approach, 2nd ed. (Wilkinson, D. G., ed.), IRL Press, Oxford, pp. 107–126.
De Block, M. and Debrouwer, D. (1993) RNA-RNA in situ hybridization using digoxigenin-labeled probes: the use of high-molecular-weight polyvinyl alcohol in the alkaline phosphatase indoxyl-nitroblue tetrazolium reaction. Anal. Biochem. 215, 86–89.
Nishioka, E., Tanaka, T., Yoshida, H., Matsumura, K., Nishikawa, S., Naito, A., et al. (2002) Mucosal addressin cell adhesion molecule 1 plays an unexpected role in the development of mouse guard hair. J. Invest. Dermatol. 119, 632–638.
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© 2005 Humana Press Inc.
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Byrne, C., Hardman, M.J. (2005). Whole-Mount Assays for Gene Induction and Barrier Formation in the Developing Epidermis. In: Turksen, K. (eds) Epidermal Cells. Methods in Molecular Biology™, vol 289. Humana Press. https://doi.org/10.1385/1-59259-830-7:127
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DOI: https://doi.org/10.1385/1-59259-830-7:127
Publisher Name: Humana Press
Print ISBN: 978-1-58829-267-4
Online ISBN: 978-1-59259-830-4
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