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BOR - Papers in Press, published online ahead of print November 27, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.008573
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Dariusz J. Skarzynski
Mamadou M. Bah
Katarzyna M. Deptula
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Submitted June 22, 2002
Returned for revision July 8, 2002
Accepted November 25, 2002

Female Reproductive Tract


Administration of a Nitric Oxide Synthase Inhibitor Counteracts Prostaglandin F2{alpha}-Induced Luteolysis in Cattle

Dariusz J. Skarzynski 1*, Jerzy J. Jaroszewski 2, Mamadou M. Bah 3, Katarzyna M. Deptula 3, Beata Barszczewska 2, Barbara Gawronska 4, William Hansel 4
1 Institute of Animal Reproduction and Food Research
2 Dep. Pharmacology, Fac.Vet.Med.Univ.Warmia & Mazury
3 Institute of Animal Reproduction and Food Research
4 Pennington Biom.Res.Center, Louisiana St.Univ. Baton Rouge

* To whom correspondence should be addressed. E-mail: skadar{at}pan.olsztyn.pl.

Abstract

The objective of this study was to determine whether a) nitric oxide (NO) is produced locally in the bovine corpus luteum (CL) and b) NO mediates prostaglandin F2{alpha} (PGF2{alpha})-induced regression of the bovine CL in vivo. The local production of NO was determined in early-I, early-II, mid, late and regressed stages of CL by determination of NADPH-d activity and the presence of inducible and endothelial NO synthase (iNOS and eNOS) immunolabelling. To determined whether inhibition of NO production counteract the PGF2{alpha}-induced regression of the CL, saline (10 ml/h; n=10) or a non-selective NOS inhibitor (N{omega}-nitro-L-arginine methyl ester dihydrochloride, L-NAME; 400 mg/h; n=9) was infused for 2 h on Day 15 of the estrous cycle into the aorta abdominalis of Holstein/Polish Black and White heifers. After 30 min of infusion, saline or cloprostenol, an analogue of PGF2{alpha} (aPGF2{alpha}; 100 mg) was injected into the aorta abdominalis of animals infused with saline or with L-NAME. NADPH-diaphorase activity was present in bovine CL, with the highest activity at mid-and late luteal stages (P<0.05). iNOS and eNOS were observed with the strongest immunolabelling in the late CL (P<0.05). Injection of aPGF2{alpha} increased nitrite/nitrate concentrations (P<0.01) and inhibited P4 secretion (P<0.05) in heifers infused with saline. Infusion of L-NAME stimulated P4 secretion (P<0.05) and concomitantly inhibited plasma concentrations of nitrite/nitrate (P<0.05). Concentrations of P4 in heifers infused with L-NAME and injected with aPGF2{alpha}were higher (P<0.05) compared to animals injected only with aPGF2{alpha}. aPGF2{alpha} shortened cycle length compared to saline (17.5 ± 0.22 days vs. 21.5 ± 0.65 days P<0.05). L-NAME blocked the luteolytic action of the aPGF2{alpha} (22.6 ± 1.07 days vs. 17.5 ± 0.22 days, P<0.05). These results suggest that NO is produced in the bovine CL. NO inhibits luteal steroidogenesis and it may by one of components of an autocrine/paracrine luteolytic cascade induced by PGF2{alpha}.



Key words: Female Reproductive Tract • Corpus luteum function • Nitric oxide • Ovulatory cycle • Progesterone


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