Immunoelectron microscopic localization of estrogen receptor on pre-mRNA containing constituents of rat uterine cell nuclei.

The localization and quantitative changes of estradiol receptor (ER) were studied by means of immunogold-electron microscope methods using a polyclonal antibody directed against an amino acid sequence representing the DNA binding site of ER, a monoclonal antibody against hnRNP core protein, and anti-DNA antibody. The uteri of normal rats in estrus and those of ovariectomized females were used. Ovariectomized rats were studied 21 days after surgery at different times after the injection of normal saline or estradiol-17 beta. The density of labeling was measured in interchromatin space, compact chromatin, nucleolus, cytoplasm, and background of epithelial cells, muscle cells, and fibroblasts. In the three types of cells ER was found mainly on extranucleolar ribonucleoprotein (RNP) fibrils. In epithelial and muscle cells the nucleolus was labeled but compact chromatin was not labeled. In epithelial cells there was a low but significant labeling of the cytoplasm. Fibroblasts exhibited a low labeling of the compact chromatin. Ovariectomy did not change these distributions. The estradiol injection increased labeling in all compartments of epithelial and muscle cells but decreased the labeling of compact chromatin of fibroblasts. These results show: (a) that ER is mainly nuclear but it is also present in the cytoplasm, (b) that ER binds to the nuclear particles containing newly synthesized RNA, and (c) that the binding to RNPs does not block the DNA binding domain of the ER.