title: Dependence on prolactin of the luteolytic effect of prostaglandin F2alpha in rat luteal cell cultures.
abstract: Luteal regression is a multistep, prolonged process, and long-term luteal cultures are required for studying it in vitro. Cell suspensions from ovaries of superovulated rats were enriched with steroidogenic cells, seeded on laminin or fibronectin, and maintained in defined medium for up to 10 days. Progesterone secretion was much lower than that of 20alpha-dihydroprogesterone, a product of 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD). Prolactin added throughout the incubation period gradually increased the percent progesterone out of total progestins to fourfold, while reducing 20alpha-HSD mRNA by 73%. Luteinizing hormone accelerated the establishment of higher percent progesterone by prolactin but by itself had no effect. Prolactin did not increase total progestin production or cytochrome P450 side-chain cleavage (P450(scc)) mRNA. Cell viability was unaffected by prolactin and/or LH. Prostaglandin F2alpha (PGF2alpha) was added 7-8 days after seeding. In prolactin-treated cells, PGF2alpha reduced steroidogenesis after 4-45 h, and at 45 h total progestins and P450(scc) mRNA were reduced by 45%. At 8-45 h PGF2alpha reduced the percent progesterone out of total progestins, and at 45 h 20alpha-HSD mRNA was doubled. In contrast, in prolactin-deprived cultures, PGF2alpha had little effect on total progestins or 20alpha-HSD mRNA but doubled P450(scc) mRNA. Phospholipase C activity was stimulated by PGF2alpha regardless of prolactin. Thus, when prolactin-treated, our cultures are a good model for mature corpora lutea challenged with PGF2alpha; the finding that without prolactin PGF2alpha has an alternative set of actions could help in identifying the signaling pathways of PGF2alpha responsible for its luteolytic effects.
authors.affiliation: Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology, Haifa 31096, Israel.
authors.forename: A
authors.initials: A
authors.lastname: Zetser
authors.forename: T
authors.initials: T
authors.lastname: Kisliouk
authors.forename: E
authors.initials: E
authors.lastname: Ivakin
authors.forename: M
authors.initials: M
authors.lastname: Lahav
mesh_terms: D015089:20-Hydroxysteroid Dehydrogenases / Q000235:genetics; D004092:20-alpha-Dihydroprogesterone / Q000096:biosynthesis; D043607:20-alpha-Hydroxysteroid Dehydrogenase; D000818:Animals; D002478:Cells, Cultured; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation / Q000187:drug effects; D015237:Dinoprost / Q000494:pharmacology*; D005260:Female; D005353:Fibronectins; D015870:Gene Expression / Q000187:drug effects; D008184:Luteal Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D007986:Luteinizing Hormone / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis; D011372:Progestins / Q000096:biosynthesis; D011388:Prolactin / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D003470:Culture Media; D005353:Fibronectins; D010716:Phosphatidylinositols; D011372:Progestins; D012333:RNA, Messenger; D004092:20-alpha-Dihydroprogesterone; D011374:Progesterone; D011388:Prolactin; D007986:Luteinizing Hormone; D004247:DNA; D015237:Dinoprost; D000242:Cyclic AMP; D015089:20-Hydroxysteroid Dehydrogenases; D043607:20-alpha-Hydroxysteroid Dehydrogenase; D002786:Cholesterol Side-Chain Cleavage Enzyme