Immuno‐electron microscopic localization of estradiol receptor in cells of male and female reproductive and non‐reproductive organs

Abstract

The localization of estradiol receptor (ER) in various tissues and their distribution in sub-cellular compartments were studied by means of immunogold-electron microscopic methods using a site-directed polyclonal antibody developed against a peptide from the DNA binding site of ER. This method was used to determine the presence and localization of ER in tissues and cells of male and female reproductive and non-reproductive organs. In the female reproductive tract, endometrial cells and the cells of the corpus luteum were found to contain ER. In non-reproductive organs of both sexes the following cell types showed significant labeling: hepatocytes, epithelial duodenal cells, striated muscle fibers, cells of the proximal convoluted tubules of the kidney, lymphocytes, neurons, and adipose cells. Alveolar epithelial cells were studied only in female specimens and were labeled by the anti-ER. Prostatic and epididymal epithelial cells were found to be labeled in the male reproductive organs. In all these cells a higher density of label was found in the nucleus, especially in the space between the clumps of compact chromatin, as was previously found in epithelial endometrial cells. These results suggest that estradiol exerts its effects through a common nuclear mechanism in cells of male and female reproductive and non-reproductive organs.

Ornithine metabolism in male and female rat kidney: mitochondrial expression of ornithine aminotransferase and arginase II

“…However, ER-␣ but not ER-␤ was found in rat kidney (17). ER has been localized in the nucleus of rat PCT cells by immunoelectron microscopy (7). In a recent study, the effects of 17␤-estradiol were analyzed by gene chip microar- Ϫ1 ].…”

Subcellular and Subnuclear Distributions of Estrogen Receptor .ALPHA. in Living Cells Using Green Fluorescent Protein and Immunohistochemistry.

“…When immunocytochemical techniques became available, it was confirmed that ERa is localized in the nuclear compartment [25,53]. Immunoelectron microscopic studies [8,14,50,53], cytochalasin B-induced enucleation procedures [49] and recent studies in living cells derived from human epithelial cell lines tagged with GFP [19] supported the nuclear localization of ERa. However, some studies showed the low but significant cytoplasmic ERa-immunoreactivity (IR) observed in various tissues [1,10,23].…”

“…Intracellular distribution of ERa has been investigated using diverse procedures which showed different results depending on the methods; cytoplasmic distribution of unliganded receptor with nuclear translocation after adding ligands [11,13], and cytoplasmic recognition with nuclear dominance [1,10,23], although the idea of the restriction in the nucleus is now most widely accepted [8,14,25,50,53]. In this study, we presented the nuclear localization of ERa with immunocytochemical procedure in fixed cells expressing endogenous ERa, Ishikawa cells derived from human endometrial adenocarcinoma [36] and MCF-7 cells from human breast carcinoma [2].…”

“…The transition of the hormonal milieu with ovariectomy [46] or estradiol (E2) treatment [8] caused no significant differences in ultrastructural localization of ERa-IR in in vivo reproductive and non-reproductive cells using immunoelectron microscopy, whereas redistribution of ERa was reported in the case of cell lines visualized with GFP and its variants after ligand treatment [19,44]. The compartmentalization of ERa in the nucleus has been studied by immunoelectron microscopy showing ERa-IR with a speckled pattern [53] in the dispersed euchromatin [14], in the margin of the condensed chromatin [50] and in the interchromatin space [8,46].…”

Immunocytochemical study of estrogen receptor activation factor (E-RAF) and the proteins that interact with nuclear estrogen receptor II (nER II) in epithelial endometrial cells, in the presence and in the absence of estradiol

“…The numerical density of the gold grains was found to be a reliable parameter to estimate the distribution of the epitope inside the cell and the changes of the density caused by experimental manipulations 1991: Vázquez-Nin et al, 1992Vázquez-Nin et al, 1993;Echeverría et al, 1994;Echeverría et al, 1999). The quantitative modifications of the abundance of E-RAF in the nucleus caused by changes in the concentration of estradiol support the observations that the decrease of estradiol concentration weakens the interaction of E-RAF with the endoplasmic reticulum letting it free to enter the nucleus (Govind et al, 2003 a, b).…”