Rapid and sensitive detection of oestrogen receptors in cells and tissue sections by autoradiography with 125I-oestradiol.
The presence of receptors for steroid hormones in individual cells and tissue sections was assessed within 4-24 h using dry mount autoradiography with radio-iodinated oestradiol. Low affinity and nonspecific binding of steroids were significantly reduced by washing the cells or sections with diluted antiserum to oestradiol. For cells of the MCF-7 cell line variations in grain density were observed, indicating that cells of the MCF-7 cell line are heterogeneous with respect to their cellular receptor concentrations of oestrogen receptors. Receptor-negative cells, such as peritoneal macrophages, did not retain oestradiol label. In tissue sections of rat and calf uterus, predominant labelling was observed on the endometrial gland cells and stroma. Oestradiol receptor binding in the uterus cytosol for both radio-iodinated and tritiated oestradiol showed the same qualitative characteristics as determined by sucrose gradient sedimentation profiles and a comparable amount of binding sites was found for both labels. The relative binding affinity of 125I-oestradiol compared to [3H]oestradiol is about 70-80%. The dry mount autoradiographic technique as presented can be used for rapid screening of heterogeneity in oestrogen receptor distribution in cells and tissue sections, since this technique reveals differences in receptor concentrations on the single cell level.
D000818:Animals; D001345:Autoradiography / Q000379:methods; D001943:Breast Neoplasms / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D002469:Cell Separation; D003600:Cytosol / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D006801:Humans; D007457:Iodine Radioisotopes; D008264:Macrophages / Q000378:metabolism; D008297:Male; D008867:Microtomy; D051381:Rats; D011919:Rats, Inbred Strains; D011960:Receptors, Estrogen / Q000032:analysis*; D014599:Uterus / Q000378:metabolism
D007457:Iodine Radioisotopes; D011960:Receptors, Estrogen; D004958:Estradiol