Prostaglandin F(2alpha) regulates the nitric oxide generating system in bovine luteal endothelial cells.
The objective of the present study was to elucidate whether luteolytic prostaglandin F(2alpha) (PGF) plays roles in regulating the nitric oxide (NO) generating system in luteal endothelial cells (LECs). Reverse transcriptase PCR, immunoblotting and immunostaining revealed the presence of PGF receptor mRNA (521 bp) and protein (64 kDa) in cultured LECs obtained from the mid-stage corpus luteum. When cultured LECs were exposed to 0.1 microM-10 microM PGF, NO production was significantly stimulated by PGF at 24 h. When LECs were exposed to 1 microM PGF for 2, 6 and 24 h, PGF did not affect the expressions of endothelial NO synthase (eNOS) mRNA and protein. On the other hand, PGF stimulated the expression of inducible NOS (iNOS) mRNA (P<0.05) and protein (P<0.05) at 2 h, but not at 6 and 24 h. By observing the conversion of [(3)C](L)-arginine to [(3)C](L)-citrulline, we found that PGF stimulated NOS activity in cultured LECs at 2 h (P<0.05). The overall findings indicate that bovine LECs are a target for PGF and that PGF stimulates iNOS expression and NOS activity in bovine LECs. Stimulation of the NO generating system and NOS activity by PGF may result in increasing local NO production followed by luteolysis.
Lee
Seung-Hyung
SH
Laboratory of Reproductive Endocrinology, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
Acosta
Tomas J
TJ
Yoshioka
Shin
S
Okuda
Kiyoshi
K
D000818:Animals; D002417:Cattle; D015237:Dinoprost / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology*; D030762:Estrous Cycle; D005260:Female; D008184:Luteal Cells / Q000166:cytology*; D008954:Models, Biological; D009566:Nitrates / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D009566:Nitrates; D009573:Nitrites; D009569:Nitric Oxide; D015237:Dinoprost; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III