Sibon O C, Cremers F F, Boonstra J, Humbel B M, Verkleij A J
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
Cell Biol Int. 1993 Jan;17(1):1-11. doi: 10.1006/cbir.1993.1001.
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.
我们通过使用地高辛标记的DNA探针，在光学显微镜水平上进行非放射性原位杂交，将表皮生长因子受体（EGF受体）的mRNA定位到A431细胞的细胞核中。在进行杂交之前，我们测试了甲醛和戊二醛固定剂。与甲醛固定相比，戊二醛产生的杂交信号扩散较少，更容易定位。因此，戊二醛被用作杂交实验中的固定剂。结果表明，EGF受体的mRNA存在于主要位于核仁周围的特定区域。提取染色质后，EGF受体mRNA的这种定位没有改变。因此可以得出结论，EGF受体的信使RNA附着在核基质上。本文还讨论了EGF受体mRNA在核仁周围定位的可能生物学作用。
