Abstract
Since presence of steroid receptors in the human placenta has been the subject of dispute, we have investigated the existence of estrogen (ER) and progesterone (PR) receptors in trophoblasts across gestational age by a variety of different techniques. Fresh human placental tissue of trimesters 1 to 3 was paraffin-embedded or snap-frozen (-80°C) and sliced (5 µm). Other tissue fragments from identical placentae were dispersed and incubated in monolayer cultures for up to 5 days. Immuno-cytochemistry (ICC) was performed for ER and PR in both trophoblast cells in culture and in whole tissue slices, using the sandwich antibody technique with subsequent horse-radish peroxidase reaction for colorization. In addition, long-term perifusion studies were conducted with explants of term placentae, using perifusion medium with estradiol (E, 2 ng/ml) and/or progesterone (P, 200 ng/ml). Perifused explants were then subjected to further ICC staining. Furthermore, RT-PCR for both ER and PR mRNA was performed for detection of the gene products in placentae of different gestational ages. Lastly, binding studies with iodine or tritium-labeled E and P were conducted on cytosol fractions. In placental sections and cultured trophoblasts, PR was clearly demonstrable in all placentae across different gestational ages. Abundant PR signal was found adjacent to the nuclei, and additionally in the dendrite-like pseudopods of syncytiotrophoblast cells. In contrast, no such staining signal was detected for the ER; this finding applied under all conditions investigated and at all gestational ages. Again, no staining for ER by ICC was detected in any tissue after perifusion with sex steroids. RT-PCR revealed no product for ER, but only for PR, in placentae across all gestational ages. Binding studies with labeled E and P showed no binding for either compound. Taken together, these observations suggest the presence of PR, but not of ER, in human placenta throughout gestation. Our failure to detect the ER does not entirely preclude the presence of this receptor in human trophoblasts, but might be attributed to a relatively low number and density of ER on these cells. Alternatively, estrogen's action on the placenta may be mediated by a different type of ER, such as by a non-classical membrane-bound receptor.
Key words
Placenta - Human - Estrogen Receptor - Progesterone Receptor - Immunocytochemistry - RT-PCR - Binding Studies
