title: Effect of endothelin-1 on bovine luteal cell function: role in prostaglandin F2alpha-induced antisteroidogenic action.
abstract: Endothelin-1 (ET-1) a vasoactive peptide, is synthesized and secreted by endothelial cells. In the bovine corpus luteum (CL), endothelial cells constitute a major proportion (53.5%) of total CL cells. This study was designed to examine the effects of ET-1 on bovine luteal cell functions and its involvement in the action of PGF2alpha. To better define the cells implicated in this process, we used CL slices, whole CL-derived cells, and steroidogenic large (LLC) and small (SLC) luteal-like cells. High affinity binding sites for ET-1 (K(d), approximately 0.3 x 10(-9)) were present in both steroidogenic luteal cells. The binding affinity of ET-1 was 3 orders of magnitude higher than that of ET-3, and a selective ETA receptor antagonist (BQ123) competed similarly to ET-1, suggesting the presence of ETA receptors. The lack of effect of ET-3 on CL-derived cells further supported this conclusion. Both basal progesterone secretion and bovine LH (5 ng/ml)-stimulated progesterone secretion from CL-derived cells were significantly inhibited by ET-1 in a dose-dependent manner, whereas preincubation of these cells with ETA receptor antagonist prevented the inhibitory effect of added ET-1. Incubation of LLC with 10(-8) M ET-1 inhibited their progesterone secretion (114.8 vs. 176.7 ng/10(5) cells-20 h; P < 0.05). On the other hand, ET-1 did not affect progesterone production from SLC despite the presence of ET-binding sites. PGF2alpha only inhibited LH-stimulated progesterone secretion by luteal slices. This antisteroidogenic effect of PGF2alpha could be prevented by the addition of a selective ETA receptor antagonist. Luteal tissue and microvascular endothelial cells isolated from bovine CL produced ET-1; in contrast, the peptide was undetectable in the culture medium or in cell extracts of either LLC or SLC. These data support the concept that ET-1 may play a paracrine regulatory role in bovine luteal function and propose a novel role for this peptide in mediating PGF2alpha-induced luteal regression.
authors.affiliation: Department of Animal Science, Faculty of Agriculture, Hebrew University of Jerusalem, Israel.
authors.forename: E
authors.initials: E
authors.lastname: Girsh
authors.forename: R A
authors.initials: RA
authors.lastname: Milvae
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: R
authors.initials: R
authors.lastname: Meidan
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003338:Corpus Luteum / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D015237:Dinoprost / Q000494:pharmacology*; D016232:Endothelins / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D011374:Progesterone / Q000378:metabolism; D013256:Steroids / Q000037:antagonists & inhibitors*
chemical_list: D016232:Endothelins; D013256:Steroids; D011374:Progesterone; D015237:Dinoprost