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ABSTRACT: We investigated whether urothelium modulates isolated rat ureter contractions.
Segments of intact and urothelium-free ureters were placed in organ baths at 37C. The contractile effects of KCl and endogenous ureteral contractile agents were recorded in the absence and presence of the cyclooxygenase inhibitors indomethacin (1 microM) or ketoprofen (10 microM). The effect of the prostacyclin analogue iloprost was tested on the KCl and agonist induced responses obtained in the presence of ketoprofen.
Without stimulation ureters were quiescent but spontaneous contractions often developed in urothelium-free ureters. Sensitivity to KCl was greater in the absence of urothelium. In intact ureters neurokinin A and vasopressin induced rhythmic contractions, whereas carbachol, norepinephrine, bradykinin and angiotensin II were inactive. In urothelium-free ureters the response to neurokinin A and vasopressin was enhanced and the other agonists, except norepinephrine, promoted contractions. In the presence of cyclooxygenase inhibitors intact ureters responded to carbachol, bradykinin and angiotensin II, and the response to neurokinin A, vasopressin and KCl increased. Responses obtained in urothelium-free ureters were not affected by the presence of cyclooxygenase inhibitors. In the presence of ketoprofen iloprost antagonized the KCl and agonist induced contractile effects in intact but not in urothelium-free ureters.
Data suggest that the urothelium prevents spontaneous contractile activity and decreases the potential excitatory effects of endogenous contractile agents on ureteral motility. The mechanism underlying this inhibitory effect appears to involve the participation of a urothelial cyclooxygenase product such as prostacyclin, which could activate the release of urothelium derived relaxing factor(s) that are as yet unknown.
The Journal of Urology 09/2007; 178(2):702-9. · 3.75 Impact Factor
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ABSTRACT: Recent studies have suggested that nitric oxide (NO) synthase (NOS) may be localized in the urothelium of the proximal part of the mammalian ureter. We investigated endogenous NO production in the proximal half of the rat ureter, localized its cellular source, characterized the NOS isoforms involved and assessed the impact of NO on ureteral motility.
Direct detection of NO production was performed on primary cultures of living rat ureteral cells with the fluorescent indicator diaminofluorescein. Cultures were incubated with the NO precursor L-arginine or the NOS inhibitors L-NAME (N-nitro-L-arginine-methyl ester) and 1400W. NOS expression was determined by immunofluorescence and Western blot analysis. The functional effects of NO donors were assessed on isolated ureters.
Significant basal NO production was demonstrated by the high fluorescence level detected in diaminofluorescein treated cell cultures. NO production was strictly limited to urothelial cells since no fluorescence was seen in smooth muscle cells. Pretreatment with L-NAME or 1400W resulted in a significant decrease in fluorescence. Constitutive and inducible NOS isoforms were detected in urothelial cultured cells and in lysates of the urothelial layer. NO donors inhibited in a concentration dependent manner the agonist induced contractile activity of isolated ureters.
These results suggest that NO production stems from the urothelium and the NO pathway inhibits contractile activity in the proximal half of the rat ureter. Hence, the nitrergic pathway may be an important target for drugs producing relaxation of the mammalian ureter.
The Journal of Urology 11/2003; 170(4 Pt 1):1409-14. · 3.75 Impact Factor
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ABSTRACT: We have examined the role of mechanical tension in myofibroblast differentiation using two in vivo rat models. In the first model, granulation tissue was subjected to an increase in mechanical tension by splinting a full-thickness wound with a plastic frame. Myofibroblast features, such as stress fiber formation, expression of ED-A fibronectin and alpha-smooth muscle actin (alpha-SMA) appeared earlier in splinted than in unsplinted wounds. Myofibroblast marker expression decreased in control wounds starting at 10 days after wounding as expected, but persisted in splinted wounds. In the second model, granuloma pouches were induced by subcutaneous croton oil injection; pouches were either left intact or released from tension by evacuation of the exudate at 14 days. The expression of myofibroblast markers was reduced after tension release in the following sequence: F-actin (2 days), alpha-SMA (3 days), and ED-A fibronectin (5 days); cell density was not affected. In both models, isometric contraction of tissue strips was measured after stimulation with smooth muscle agonists. Contractility correlated always with the level of alpha-SMA expression, being high when granulation tissue had been subjected to tension and low when it had been relaxed. Our results support the assumption that mechanical tension is crucial for myofibroblast modulation and for the maintenance of their contractile activity.
American Journal Of Pathology 10/2001; 159(3):1009-20. · 4.89 Impact Factor
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ABSTRACT: Non-steroidal anti-inflammatory drugs (NSAIDs) are currently considered a first-line treatment of renal colic. Their action has been ascribed to the inhibition of renal prostaglandin synthesis, which decreases renal blood flow and diuresis, and consequently lowers the pressure in the renal pelvis and ureter. However, the effects of NSAIDs on induced contractions of ureteral smooth muscle have received little attention. Also, there is a lack of clinically relevant spasmolytic drugs for the ureter. Therefore, we studied the influence of the non-selective cyclooxygenase (COX) inhibitor diclofenac, a NSAID drug customarily used in the treatment of renal colic, and of NS-398, a selective COX-2 inhibitor, on induced contractions of the pig ureter. Serotonin (0.1-30 microM), norepinephrine (0.1-30 microM) and neurokinin A (0.03-10 microM) induced reproducible concentration-dependent contractions, which were inhibited by diclofenac and NS-398 (10-300 microM) in a concentration-dependent manner. The sensitivity of neurokinin A-induced contractions to diclofenac was 3-4 times greater than that of the amines. Depending on the concentration, inhibition ranged between 25 and 96% of the initially induced contractile activity. In the presence of inhibitors, supramaximal concentrations of agonists were unable to trigger recuperation of the initially induced contractions. Prostaglandin F2alpha did not reverse the effect of diclofenac on agonist-induced contractions. Removal of diclofenac or NS-398 from the organ baths showed that the inhibition was totally reversible. Thus, the non-selective COX inhibitor diclofenac and the selective COX-2 inhibitor NS-398 are almost equipotent in reducing agonist-induced contractions in the isolated porcine ureter. Although the clinical relevance of this spasmolytic effect remains to be demonstrated, the data suggest that patients suffering from renal colic may benefit not only from the anti-diuretic and analgesic effects of diclofenac, but also from its potential spasmolytic properties. Moreover, selective COX-2 inhibitors may have clinical potential, as they may cause fewer side effects.
Urological Research 01/2001; 28(6):376-82. · 1.23 Impact Factor
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ABSTRACT: The portal vein has spontaneous and agonist-induced contractile activities and whether sepsis alters these two types of contractile activities is unknown.
To study the effect of sepsis on the spontaneous contractile activity and the contractile responses to norepinephrine (NE), angiotensin II (AT(II)), and neurokinin B (NKB) in the rat portal vein (RPV), we performed a cecal ligature and puncture (CLP) 24 h before RPV isolation.
CLP decreased the spontaneous activity and induced hyporesponsiveness to AT(II) and NKB. The vascular failure was correlated to the severity of sepsis. In contrast, the reactivity to NE was not altered. Although inducible NO synthase was detected in RPV isolated from CLP rats, NO synthase inhibitors did not restore either the responsiveness to AT(II) and NKB or the spontaneous activity. Additionally, hyporesponsiveness to AT(II) and NKB was not modified by indomethacin.
CLP decreases the spontaneous activity of the RPV as well as the contractile responses to AT(II) and NKB. The vascular failure is correlated to the severity of sepsis. The reactivity to NE is not altered in this model. Neither NO nor prostaglandins are responsible for the vascular abnormalities observed during CLP.
Journal of Hepatology 01/2001; 33(6):933-40. · 9.26 Impact Factor
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ABSTRACT: To investigate the systemic and hepatic reactivity to norepinephrine (NE) during chronic endotoxemia, we measured the reactivity of the drug in pigs infused with endotoxin (End, 160 ng/kg/min) during an 18-h period. At the end of the experiments, the hepatic vessels were removed to test the hepatic vascular reactivity in vitro. The pressive response to NE did not change over time in control (Ctrl) pigs, but endotoxin infusion decreased the response at t = 11 and 17 h. The reactivity of the portal vein blood flow did not change in Ctrl pigs but was significantly increased in End pigs at t = 5 h. Finally, endotoxin decreased the contractile response to NE only in transversal strips of portal veins isolated from End pigs. Thus, in this model, the decreased pressive response to NE develops over time, but the modifications of the hepatic vascular reactivity remain minor.
Shock 11/1999; 12(4):294-9. · 2.85 Impact Factor
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ABSTRACT: In pig and humans, whose kidneys have a multi-calyceal collecting system, the initiation of ureteral peristalsis takes place in the renal calyces. In the pig and human ureter, recent evidence suggests that nitric oxide (NO) is an inhibitory mediator that may be involved in the regulation of peristalsis. This study was designed to assess whether the NO synthase/NO/cyclic GMP pathway modulates the motility of pig isolated calyceal smooth muscle. Immunohistochemistry revealed a moderate overall innervation of the smooth muscle layer, and no neuronal or inducible NO synthase (NOS) immunoreactivities. Endothelial NOS immunoreactivities were observed in the urothelium and vascular endothelium, and numerous cyclic GMP-immunoreactive (-IR) calyceal smooth muscle cells were found. As measured by monitoring the conversion of L-arginine to L-citrulline, Ca(2+)-dependent NOS activity was moderate. Assessment of functional effects was performed in tissue baths and showed that NO and SIN-1 decreased spontaneous and induced contractions of isolated preparations in a concentration-dependent manner. In strips exposed to NO, there was a 10-fold increase of the cyclic GMP levels compared with control preparations (P < 0.01). It is concluded that a non-neuronal NOS/NO/cyclic GMP pathway is present in pig calyces, where it may influence motility. The demonstration of cyclic GMP-IR smooth muscle cells suggests that NO acts directly on these cells. This NOS/NO/cyclic GMP pathway may be a target for drugs inhibiting peristalsis of mammalian upper urinary tract. Neurourol. Urodynam. 18:673-685, 1999.
Neurourology and Urodynamics 02/1999; 18(6):673-85. · 2.96 Impact Factor
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ABSTRACT: Acetylcholine promotes paradoxical sleep (PS), but the role of noradrenaline in this stimulation is controversial. The relationship between cholinergic and noradrenergic systems in the production of PS was investigated in the rat implanted on a continuous basis for sleep recordings. Stimulation of PS was obtained with microinjections of carbachol (1 microgram) into the pontine reticular formation. In the presence of the alpha 2-agonist clonidine (5 micrograms/kg, IP), the carbachol activation of PS was abolished. This stimulation also disappeared when the animals were pretreated with alpha-methyl-paratyrosine (150 mg/kg, IP), an inhibitor of catecholamine synthesis. Thus, carbachol stimulation appeared inefficient when brain noradrenergic activation was decreased. This observation supports the view that the realization of PS by the cholinergic system requires a certain level of noradrenergic activity.
Pharmacology Biochemistry and Behavior 06/1994; 48(1):93-100. · 2.53 Impact Factor
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ABSTRACT: Active neurokinin (NK) receptors were visualized with 5-nm colloidal gold-protein-substance P (GPSP) and -senktide (GPSenk) complexes on vascular tissues. Electron micrographs of pig coronary strips incubated with GPSP showed gold particles either bound to the plasmalemma or inside intracytoplasmic vesicles of endothelial cells. Preincubation with SP or the NK1 receptor antagonist L-703606 prevented GPSP marking. No gold particles were seen after incubation with GPSenk. On coronary strips in vitro, which had been precontracted with U46619, GPSP induced relaxations similar to those produced by equimolar concentrations of SP, both relaxations being inhibited by L-703606. Analogous to senktide, GPSenk was totally inactive on arterial strips. Incubation of rat portal veins with GPSenk showed gold particles bound to the plasmalemma or inside intracytoplasmic vesicles of smooth muscle cells. Preincubation with senktide or the NK3 receptor antagonist R-820 prevented GPSenk marking. On portal veins in vitro GPSenk induced contractions similar to those induced by equimolar concentrations of senktide or NKB; these effects were inhibited by R-820. Our results show that colloidal gold-protein complexes present biological activity and selectivity similar to those of their respective native ligand and detect the presence of active receptors; in addition, they suggest the presence of NK-receptor-mediated endocytosis in endothelial cells of coronary artery and in smooth muscle cells of the portal vein.
Journal of Vascular Research 36(1):59-67. · 2.65 Impact Factor
