title: Effects of a nitric oxide donor and nitric oxide synthase inhibitors on luteinizing hormone-induced ovulation in the ex-vivo perfused rat ovary.
abstract: The aim of this study was to investigate the role of nitric oxide (NO) in ovulation and ovarian steroidogenesis by the use of NO synthase (NOS) inhibitors and an NO donor administrated to the luteinizing hormone (LH)-stimulated ex-vivo perfused pre-ovulatory rat ovary. The ovaries were stimulated with LH (0.2 microgram/ml) alone or in combination with the phosphodiesterase inhibitor IBMX (200 micromol/l). The presence of both endothelial NOS (eNOS) and inducible NOS (iNOS) in the perfused rat ovary were detected by immunoblotting and a clear increase in amount of iNOS protein was seen after LH+IBMX stimulation. The addition of a non-selective NOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA; 300 micromol/l), to the perfusate significantly decreased ovulation numbers (median = 4. 0, range = 1-14) as compared with LH + IBMX stimulated control (12.0, 6-17). In contrast, an inhibitor with relative selectivity towards iNOS, aminoguanidine bicarbonate (AG, 300 micromol/l and 1 mmol/l), did not change the ovulation rate (11.5, 6-18 and 11.0, 7-15 respectively). In perfusions with only LH, a lower ovulation rate was seen but with similar effects (0.0, 0-8 for L-NMMA; 7.5, 3-12 for control and 7.0, 1-15 for AG 300 micromol/l). The administration of an NO donor, spermine NONOate, resulted in similar ovulation numbers as in LH-stimulated controls. The NO inhibitors did not affect steroid concentrations in the perfusion media, while 100 micromol/l NONOate increased progesterone production.
authors.affiliation: Department of Obstetrics and Gynecology, Göteborg University, Sahlgrenska University Hospital, S-413 45 Göteborg, Sweden.
authors.forename: K
authors.initials: K
authors.lastname: Mitsube
authors.forename: M
authors.initials: M
authors.lastname: Mikuni
authors.forename: M
authors.initials: M
authors.lastname: Matousek
authors.forename: M
authors.initials: M
authors.lastname: Brännström
mesh_terms: D000818:Animals; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D007986:Luteinizing Hormone / Q000627:therapeutic use*; D009569:Nitric Oxide / Q000037:antagonists & inhibitors*; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D010053:Ovary / Q000187:drug effects*; D010062:Ovulation Induction / Q000379:methods*; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D020030:Nitric Oxide Donors; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D007986:Luteinizing Hormone; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496318:Nos2 protein, rat; C496316:Nos3 protein, rat