Localisation of EGF-Receptor mRNA in the nucleus of A431 cells by light microscopy.

Authors

  • Ody C. M. Sibon,

    Corresponding author
    1. Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Tel: 31-30-533449, Fax: 31-30-513655
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  • Fons F. M. Cremers,

    1. Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Tel: 31-30-533449, Fax: 31-30-513655
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  • Johannes Boonstra,

    1. Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Tel: 31-30-533449, Fax: 31-30-513655
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  • Bruno M. Humbel,

    1. Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Tel: 31-30-533449, Fax: 31-30-513655
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  • Arie J. Verkleij

    1. Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Tel: 31-30-533449, Fax: 31-30-513655
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Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands. Tel: 31-30-533449, Fax: 31-30-513655

Abstract

We have localized the mRNA of the epidermal growth factor receptor (EGF-receptor) in nuclei of A431 cells by non-radioactive in situ hybridization at the light microscopical level using digoxigenin-labelled DNA probes. Both formaldehyde and glutaraldehyde fixations were tested before the hybridization was performed. Glutaraldehyde, compared with formaldehyde fixation, gives a less diffuse hybridization signal, which is easier to localize. Therefore, glutaraldehyde was used as a fixative in the hybridization experiments. It is demonstrated that the mRNA of the EGF-receptor is present in restricted domains mainly located around the nucleolus. This location of the EGF-receptor mRNA was unaltered after extraction of chromatin. Therefore it is concluded that the messenger RNA of the EGF-receptor is attached to the nuclear matrix. A possible biological role for the location of mRNA of the EGF-receptor around the nucleolus is discussed.

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