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We have recently provided evidence for the desensitization of oxytocin receptors in human myometrial cells. In the present study, we have investigated the possible mechanisms by which oxytocin (OT) might regulate OT receptor density. The steady state level of OT binding in cultured myometrial cells was 220 x 10(3) binding sites/ cell, but this was time-dependently reduced to 27 x 10(3) sites/cell by exposure to OT for up to 20 h. Similarly, OT exposure decreased the binding of OT to cell membranes. In contrast, Western blotting data showed that the total amount of OT receptor protein was not affected by OT treatment for up to 48 h. Flow cytometry experiments demonstrated that OT receptors are not internalized during prolonged exposure of the cells to OT. However, RNase protection assays and Northern analysis showed that OT receptor mRNA was reduced by OT treatment to reach a new low steady state level with a time course similar to that of the disappearance of cell surface OT binding sites. Possible mechanisms involved in mRNA down-regulation include transcriptional suppression and destabilization of mRNA by RNA binding proteins.

Original publication

DOI

10.1677/joe.0.1540007

Type

Journal article

Journal

J Endocrinol

Publication Date

07/1997

Volume

154

Pages

7 - 18

Keywords

Blotting, Northern, Blotting, Western, Cells, Cultured, Down-Regulation, Female, Flow Cytometry, Gene Expression Regulation, Humans, Myometrium, Oxytocin, Pregnancy, Protein Binding, RNA, Messenger, Receptors, Oxytocin