Identification of nitric oxide synthase in human and bovine oviduct.
Nitric oxide synthase (NOS) is responsible for the biological production of nitric oxide (NO) in several organs. NOS activity has also been localized in the reproductive tract, although direct evidence for its presence in the human or bovine oviduct is still lacking. In the present study, four different techniques were used to identify the presence of NOS activity in human (n = 11) and bovine (n = 9) oviduct: (i) conversion of [3H]-L-arginine to [3H]-L-citrulline; (ii) production of nitrite/nitrate (NO2/NO3; stable NO metabolites); (iii) identification of NADPH-diaphorase activity; and (iv) immunostaining with antiserum to endothelial NOS. Cytosolic extracts from human ampullary segments of the Fallopian tube, obtained from post-partum patients (n = 4), converted [3H]-L-arginine to [3H]-L-citrulline (21.0 +/- 8.8 fmol/mg protein/min). This conversion rate was significantly (P < 0.05) reduced in the presence of either EDTA or N-monomethyl-L-arginine monoacetate (L-NMMA), an inhibitor of NOS activity. When bovine (n = 3) ampullary segments were incubated for 36 h in Hanks' balanced salt solution, the concentration of NO2/NO3 in the medium was increased (P < 0.05) if segments were pretreated with lipopolysaccharide (LPS; an inducer of inducible NOS), but not after treatment with LPS + L-NMMA. Additionally, epithelial cells cultured from ampullary segments showed positive staining both for NADPH-diaphorase activity and with antiserum to endothelial NOS. The results of the present study provide direct evidence for the presence of both the Ca(2+)-dependent constitutive form of NOS, as well as the inducible form of NOS activity in human and bovine oviduct. Since the oviduct plays a key role in the reproductive process, it is possible that the two forms of NOS may be involved in the physiological regulation of oviduct function.
Rosselli
M
M
Department of Obstetrics and Gynaecology, University Hospital Zurich, Switzerland.
Dubey
R K
RK
Rosselli
M A
MA
Macas
E
E
Fink
D
D
Lauper
U
U
Keller
P J
PJ
Imthurn
B
B
D000818:Animals; D001120:Arginine / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D002956:Citrulline / Q000096:biosynthesis; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000378:metabolism; D005187:Fallopian Tubes / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D009252:NADPH Dehydrogenase / Q000378:metabolism; D009566:Nitrates / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D009573:Nitrites / Q000378:metabolism
D009566:Nitrates; D009573:Nitrites; D002956:Citrulline; D001120:Arginine; D019001:Nitric Oxide Synthase; D009252:NADPH Dehydrogenase