Human Reproduction, Vol. 14, No. 10, 2537-2543,
October 1999
© 1999 European Society of Human Reproduction and Embryology
Effects of a nitric oxide donor and nitric oxide synthase inhibitors on luteinizing hormone-induced ovulation in the ex-vivo perfused rat ovary
Department of Obstetrics and Gynecology, Göteborg University, Sahlgrenska University Hospital, S-413 45 Göteborg, Sweden
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 µg/ml) alone or in combination with the phosphodiesterase inhibitor IBMX (200 µmol/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, NG-monomethyl-L-arginine (L-NMMA; 300 µmol/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 µmol/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 µmol/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 µmol/l NONOate increased progesterone production.
Key words: nitric oxide/NOS inhibitor/ovarian perfusion/ovulation/rat
1 To whom correspondence should be addressed
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