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

Cell Biol Int. 1993 Jan;17(1):1-11. doi: 10.1006/cbir.1993.1001.

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.

MeSH terms

  • Animals
  • Carcinoma, Squamous Cell
  • Cell Nucleolus / chemistry
  • Cell Nucleolus / ultrastructure
  • Cell Nucleus / chemistry*
  • Cell Nucleus / ultrastructure
  • DNA Probes
  • DNA, Neoplasm / analysis
  • DNA, Neoplasm / genetics
  • ErbB Receptors / chemistry*
  • Formaldehyde / pharmacology
  • Glutaral / pharmacology
  • Humans
  • In Situ Hybridization
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics
  • Tumor Cells, Cultured / chemistry*
  • Tumor Cells, Cultured / ultrastructure

Substances

  • DNA Probes
  • DNA, Neoplasm
  • RNA, Messenger
  • Formaldehyde
  • ErbB Receptors
  • Glutaral