Localisation of EGF-receptor mRNA in the nucleus of A431 cells by light microscopy.
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.
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
O C
OC
Sibon
F F
FF
Cremers
J
J
Boonstra
B M
BM
Humbel
A J
AJ
Verkleij
D000818:Animals; D002294:Carcinoma, Squamous Cell; D002466:Cell Nucleolus / Q000737:chemistry / Q000648:ultrastructure; D002467:Cell Nucleus / Q000737:chemistry* / Q000648:ultrastructure; D015342:DNA Probes; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D066246:ErbB Receptors / Q000737:chemistry*; D005557:Formaldehyde / Q000494:pharmacology; D005976:Glutaral / Q000494:pharmacology; D006801:Humans; D017403:In Situ Hybridization; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D014407:Tumor Cells, Cultured / Q000737:chemistry* / Q000648:ultrastructure
D015342:DNA Probes; D004273:DNA, Neoplasm; D012333:RNA, Messenger; D005557:Formaldehyde; D066246:ErbB Receptors; D005976:Glutaral