Interaction of mouse placental lactogens and androgens in regulating progesterone release in cultured mouse luteal cells.
Pituitary hormones are essential for the maintenance of the corpus luteum in the pregnant mouse during the first half of gestation. Thereafter, hormones from the placenta take over the luteotropic role of the pituitary hormones. Mouse placental lactogen-I (mPL-I) and mPL-II, two PRL-like hormones produced in the placenta, are probably necessary for the maintenance of the corpus luteum in the latter half of pregnancy. A culture system of luteal cells from pregnant mice was developed to investigate the role of hormones from the placenta that may be important for the function of the corpus luteum. Mice were killed on days 10, 14, and 18 of pregnancy, and the corpora lutea were excised from the ovaries and digested in 0.1% collagenase, 0.002% DNase for 1 h. The resulting luteal cell suspension was plated onto 96-well plates coated with fibronectin (1 x 10(5) cells/well) and cultured for 1-3 days. Medium was changed daily. The cells were treated with various concentrations and combinations of mPL-I, mPL-II, mouse PRL, androstenedione, dihydrotestosterone, 17beta-estradiol (E2), testosterone, hydroxyflutamide, cycloheximide, actinomycin D, and fadrozole to study the effects of these different treatments on progesterone (P4) production. The three lactogens (mPL-I, mPL-II, and mouse PRL) all stimulated the release of P4 from the luteal cells. The potency of the lactogens was similar and did not depend on the stage of pregnancy at which the luteal tissue was obtained. However, the responsiveness of the cells to all hormone-stimulated P4 release was gradually reduced the later in pregnancy the tissue was collected. Androgens also stimulated the release of P4 from the luteal cells, and when administered together, the lactogens and the androgens acted synergistically to stimulate P4 release. The androgens acted directly but not through conversion to E2, as determined by the findings that 1) the effects of the androgens could not be reproduced by E2 administration, 2) nonaromatizable androgen dihydrotestosterone was as effective as aromatizable androgens, and 3) aromatase inhibitor did not prevent the action of the androgens to stimulate the P4 release. The effect of the androgens on the P4 release was rapid, occurring within 15 min of hormone administration. It was not prevented by inhibitors of protein and RNA synthesis, and the intracellular androgen receptor antagonist hydroxyflutamide did not affect the androgen action. Therefore, the androgen effects were not mediated through the intracellular androgen receptor and de novo protein synthesis was not needed for androgen-stimulated P4 release.
Thordarson
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G
Department of Biology, Sinsheimer Laboratories, University of California, Santa Cruz 95064, USA.
Galosy
S
S
Gudmundsson
G O
GO
Newcomer
B
B
Sridaran
R
R
Talamantes
F
F
D059002:Androgen Receptor Antagonists; D000728:Androgens / Q000494:pharmacology* / Q000502:physiology; D000735:Androstenedione / Q000494:pharmacology; D000818:Animals; D047072:Aromatase Inhibitors; D002417:Cattle; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D013196:Dihydrotestosterone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D017316:Fadrozole / Q000494:pharmacology; D005260:Female; D005485:Flutamide / Q000031:analogs & derivatives / Q000494:pharmacology; D008184:Luteal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D010928:Placental Lactogen / Q000494:pharmacology* / Q000502:physiology; D011247:Pregnancy; D011374:Progesterone / Q000378:metabolism*; D011388:Prolactin / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011817:Rabbits; D013739:Testosterone / Q000494:pharmacology
D059002:Androgen Receptor Antagonists; D000728:Androgens; D047072:Aromatase Inhibitors; D004791:Enzyme Inhibitors; D004965:Estrogen Antagonists; D011500:Protein Synthesis Inhibitors; D013196:Dihydrotestosterone; C014290:hydroxyflutamide; D013739:Testosterone; D000735:Androstenedione; D011374:Progesterone; D004958:Estradiol; D005485:Flutamide; D011388:Prolactin; D010928:Placental Lactogen; D003513:Cycloheximide; D017316:Fadrozole