PDF
ePub
Citation
Print
Abstract
Purpose
Nitric oxide synthase (NOS) is an important enzyme in the production of nitric oxide (NO). The constitutive type (cNOS) is expressed in the normal physiologic state, and the inducible type (iNOS) in expressed in the active immune state. cNOS is divided into an endothelial type (eNOS) and a neuronal type (nNOS). eNOS affects blood vessels, while nNOS affects nerve fibers. In the present study, we evaluated the expression of eNOS and nNOS in rat bladders with short-term partial outlet obstructions. We presupposed that NO is responsible for prolonged micturition problems after partial outlet obstruction.
Materials and Methods
Specific pathogen-free Sprague-Dawley rats weighing 250-300g were used for the study. Individual bladders were obtained from sham-operated control rats (n=5) and from experimental rats at 12 hours and 1, 2, 3, and 7 days after partial urethral obstruction (n=25). eNOS and nNOS were detected using immunochemical staining and analyzed with confocal microscopy and an image analyzer.
Results
eNOS and nNOS expression were detected in both the control group and in the group with partial outlet obstruction. The expression of eNOS showed a sharp increase at 3 days after obstruction and returned to normal at 7 days. The expression of nNOS was not significantly different between the two groups.
REFERENCES
1.Abrams P. Describing bladder storage function: overactive bladder syndrome and detrusor overactivity. Urology. 2003. 62(5 Suppl 2):28–37.
2.Ho MH., Bhatia NN., Khorram O. Physiologic role of nitric oxide and nitric oxide synthase in female lower urinary tract. Curr Opin Obstet Gynecol. 2004. 16:423–9.
3.Bennett BC., Kruse MN., Roppolo JR., Flood HD., Fraser M., de Groat WC. Neural control of urethral outlet activity in vivo: role of nitric oxide. J Urol. 1995. 153:2004–9.
4.Hallen K., Gustafsson LE., Wiklund NP. Nerve-induced release of nitric oxide from the rabbit corpus cavernosum is modulated by cyclic guanosine 3',5'-monophosphate. Neuroscience. 2005. 133:169–74.
5.Lemack GE., Burkhard F., Zimmern PE., McConnell JD., Lin VK. Physiologic sequelae of partial infravesical obstruction in the mouse: role of inducible nitric oxide synthase. J Urol. 1999. 161:1015–22.
6.Persson K., Poljakovic M., Johansson K., Larsson B. Morphological and biochemical investigation of nitric oxide synthase and related enzymes in the rat and pig urothelium. J Histochem Cytochem. 1999. 47:739–50.
7.Furchgott RF., Zawadzki JV. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980. 288:373–6.
8.Forstermann U., Pollock JS., Tracey WR., Nakane M. Isoforms of nitric-oxide synthase: purification and regulation. Methods Enzymol. 1994. 233:258–64.
9.Förstermann U., Closs EI., Pollock JS., Nakane M., Schwarz P., Gath I, et al. Nitric oxide synthase isozymes. Characterization, purification, molecular cloning, and functions. Hypertension. 1994. 23:1121–31.
10.Huang A., Palmer LS., Hom D., Valderrama E., Trachtman H. The role of nitric oxide in obstructive nephropathy. J Urol. 2000. 163:1276–81.
11.Whittle BJ. Nitric oxide in physiology and pathology. Histo-chem J. 1995. 27:727–37.
12.Moncada S., Palmer RM., Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991. 43:109–42.
13.Smet PJ., Jonavicius J., Marshall VR., de Vente J. Distribution of nitric oxide synthase-immunoreactive nerves and identification of the cellular targets of nitric oxide in guinea-pig and human urinary bladder by cGMP immunohistochemistry. Neuroscience. 1996. 71:337–48.
14.Saito M., Miyagawa I. N (G)-nitro-L-arginine methylester, a nitric oxide synthase inhibitor, diminishes apoptosis induced by ischemia-reperfusion in the rat bladder. Neurourol Urodyn. 2002. 21:566–71.
15.Theobald RJ Jr. Differing effects of N (G)-monomethyl L-arginine and 7-nitroindazole on detrusor activity. Neurourol Urodyn. 2003. 22:62–9.
16.Masuda H., Okuno T., Suzuki M., Kihara K., Goto M., Azuma H. Different distribution of nitric oxide synthase and soluble guanylyl cyclase activities in the detrusor and proximal urethra of the rabbit. J Urol. 2002. 168:2286–90.
17.Fathian-Sabet B., Bloch W., Klotz T., Niggemann S., Jacobs G., Addicks K, et al. Localization of constitutive nitric oxide synthase isoforms and the nitric oxide target enzyme soluble guanylyl cyclase in the human bladder. J Urol. 2001. 165:1724–9.
18.Nitsch DD., Ghilardi N., Mühl H., Nitsch C., Brune B., Pfeil-schifter J. Apoptosis and expression of inducible nitric oxide synthase are mutually exclusive in renal mesangial cells. Am J Pathol. 1997. 150:889–900.
Fig. 1.
Expression of endothelial nitric oxide synthase (eNOS) (red, Texas Red goat anti-rabbit antibody, 1:200) and neuronal NOS (nNOS) (green, anti-mouse IgG FITC, 1:200) (x400). (A) Control, (B) 12 hours after partial bladder outlet obstruction, (C) 3 days after partial bladder outlet obstruction, (D) 7 days after partial bladder outlet obstruction.
Table 1.
Changes in bladder weights
|
Bladder weights (mg) |
||
|---|---|---|
| Control | Partial bladder outlet obstruction | |
| 12 hours | 112.4±4.8 | 115.6±3.6 |
| 1 day | 117.7±5.5 | 118.2±4.3 |
| 2 days | 116.4±4.8 | 134.6±2.6 |
| 3 days | 115.3±4.6 | 158.7±11.4 |
| 7 days | 115.7±3.2 | 477.2±23.6* |
Table 2.
Changes in endothelial nitric oxide synthase expression
|
Gray scale of endothelial nitric oxide synthase |
||
|---|---|---|
| Control | Partial bladder outlet obstruction | |
| 12 hours | 66.40±2.82 | 45.80±2.35 |
| 1 day | 64.32±1.44 | 67.60±2.35 |
| 2 days | 54.36±1.65 | 69.56±5.88 |
| 3 days | 55.08±2.36 | 82.44±4.95* |
| 7 days | 55.87±2.07 | 65.67±5.46 |
XML Download