Progesterone receptor messenger ribonucleic acid in the primate corpus luteum during the menstrual cycle: possible regulation by progesterone.
In classical target tissues, progesterone (P) down-regulates its own receptor, yet in the primate corpus luteum, progesterone receptors (PRs) exist within a very high local P milieu. The percentage of luteal cells staining PR-positive by immunocytochemistry is highest at the midluteal phase of the menstrual cycle during the period of peak serum P. To investigate the regulation of luteal PRs, we developed a solution hybridization/ribonuclease protection assay for the analysis of PR messenger RNA (mRNA) in macaque corpora lutea (n = 3-4/group). A 332-basepair fragment of the macaque PR complementary DNA corresponding to the hormone-binding region was used as a template for riboprobe production; the specific hybridization of this riboprobe with PR mRNA was confirmed with Northern analysis. P regulation of luteal PR mRNA was investigated by administering trilostane, a 3 beta-hydroxysteroid dehydrogenase inhibitor, to female rhesus macaques beginning on day 6 or 7 of the luteal phase, which reduced serum P until the time of lutectomy. By 18 h after trilostane treatment, luteal PR mRNA levels were significantly elevated compared to untreated control values (mean +/- SEM, 2.0 +/- 0.4 vs. 0.7 +/- 0.3; P < 0.05). Reduction in P levels for 4 days after trilostane administration decreased luteal PR mRNA levels compared with control values (0.50 +/- 0.02 vs. 1.1 +/- 0.2; P < 0.05). To characterize changes in PR mRNA during the lifespan of the corpus luteum, mRNA levels in luteal tissues from the early, mid-, mid-late, and late luteal phases were determined. PR mRNA levels were lowest during the early luteal phase and increased (P < 0.05) 3-fold by the mid-late luteal phase; this higher PR mRNA level was maintained throughout the remainder of the luteal phase. These data indicate that P or a metabolite may acutely regulate primate luteal PR mRNA in a manner consistent with PR regulation in classical P target tissues. In contrast, PR mRNA levels parallel increases in P and PR-positive luteal cells during the early, mid-, and mid-late portions of the luteal phase. High PR mRNA levels are maintained during luteal regression as P and the percentage of PR-positive cells decline, suggesting that PR and PR mRNA are regulated in an asynchronous manner during the lifespan of the corpus luteum in the menstrual cycle.
Duffy
D M
DM
Division of Reproductive Sciences, Oregon Regional Primate Research Center, Beaverton 97006, USA.
Stouffer
R L
RL
D015096:3-Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003338:Corpus Luteum / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary; D013196:Dihydrotestosterone / Q000031:analogs & derivatives / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation; D006107:Granulosa Cells / Q000378:metabolism*; D008253:Macaca mulatta; D008597:Menstrual Cycle / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000097:blood / Q000378:metabolism / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D011980:Receptors, Progesterone / Q000096:biosynthesis*
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D011980:Receptors, Progesterone; D013196:Dihydrotestosterone; D011374:Progesterone; D015096:3-Hydroxysteroid Dehydrogenases; C009954:trilostane