Abstract
In situ hybridization techniques enable gene expression to be monitored in individual cells (Figure 18.1). Methods for use with the light microscope are as sensitive as northern blot analysis for detecting mRNAs that accumulate equally in all cells, and more sensitive than northern analysis for detecting mRNAs that are present in only a small population of cells within an organ (Langdale et al. 1988a,b; Martineau and Taylor 1986). The following protocol has been used most frequently to monitor gene expression in maize leaf tissue. By varying tissue fixation conditions, all organs of the maize plant have been analyzed. The method uses paraffin-embedded sections and 35S-radiolabeled or digoxigenin nonradiolabeled riboprobes. The use of nonradiolabeled probes is a fairly recent advance and at this stage they appear to be less sensitive than radiolabeled probes. However, the method can undoubtedly be improved with further use and optimization. Tissue that is fixed and embedded as described below may also be used for immunocytochemistry. Standard methods of microscopy and photomicroscopy are used for signal detection. See appropriate chapters in this volume for details.
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References
Cox, KH, Deleon, DV, Angerer, LM and Angerer, RC (1984) Detection of mRNAS in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev. Biol. 101485–502
Jackson D (1992) In situ hybridization in plants. In Gurr SJ, Bowles DJ, McPherson MJ (eds) Molecular Plant Pathology: A Practical Approach, Volume 1 IRC Press, Oxford, pp 163–174
Langdale JA, Zelitch I, Miller E, Nelson T (1988a) Cell position and light influence C4 versus C3 patterns of photosynthetic gene expression in maize. EMBO J 7: 3643–3651
Langdale JA, Rothermel BA, Nelson T (1988b) Cellular patterns of photosynthetic gene expression in developing maize leaves. Genes Dev 2: 106–115
Martineau B, Taylor WC (1986) Cell-specific photosynthetic gene expression in maize determined using cell separation techniques and hybridization in situ. Plant Physiol 82: 613–618
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© 1994 Springer-Verlag New York, Inc.
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Langdale, J.A. (1994). In situ Hybridization. In: Freeling, M., Walbot, V. (eds) The Maize Handbook. Springer Lab Manuals. Springer, New York, NY. https://doi.org/10.1007/978-1-4612-2694-9_18
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DOI: https://doi.org/10.1007/978-1-4612-2694-9_18
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