Walter F. Thon

Immunolocalization of Multidrug Resistance Protein 5 in the Human Genitourinary System

PURPOSE The intracellular messenger cyclic guanosine monophosphate (cGMP) has an important role in regulating smooth muscle tone. An increase in intracellular cGMP levels is a prerequisite for penile erection. Inhibition of cGMP degradation by cGMP specific phosphodiesterase 5 has been used for treating erectile dysfunction. In addition to degradation by phosphodiesterase, cGMP is exported from cells by multidrug resistance protein 5 (MRP5), also called ABCC5, which we recently identified as an adenosine triphosphate dependent export pump for cGMP. MRP5 is potently inhibited by substances known as phosphodiesterase inhibitors, including sildenafil and trequinsin. Therefore, we analyzed whether MRP5 is expressed in tissues of the human genitourinary system and whether MRP5 and phosphodiesterase 5 proteins are localized in the same cell types. MATERIALS AND METHODS Localization of MRP5 and phosphodiesterase 5 was analyzed by immunofluorescence microscopy in cryosections of various tissues of the human genitourinary system. RESULTS MRP5 and phosphodiesterase 5 were co-expressed in smooth muscle cells of the corpus cavernosum, ureter, urethra and bladder. In addition, MRP5 and phosphodiesterase 5 were localized in epithelial cells of the mucosa in the ureter and urethra, and in blood vessels of the lamina propria. CONCLUSIONS The co-expression of MRP5 and phosphodiesterase 5 in smooth muscle cells of the genitourinary system indicates 2 distinct pathways for cGMP removal. Thus, MRP5 inhibition represents a new approach for enhancing cGMP levels in smooth muscle cells and developing drugs for erectile dysfunction.

Prognostic Factors for the Postoperative Outcome of Penile Venous Surgery for Venogenic Erectile Dysfunction

Since penile venous surgery is usually associated with a poor postoperative outcome, a study was done to evaluate possible prognostic factors for this procedure. A total of 77 patients with erectile dysfunction underwent ligation of all dorsal penile veins and resection of the deep dorsal penile vein for venous incompetence. In all patients a comprehensive evaluation was done preoperatively. All patients did not respond to pharmacotherapy and had a venous leak. After a followup of 6 months, patients were classified as having full spontaneous erections, failure and response to pharmacotherapy. Of the 77 patients 31 (40.3%) had full spontaneous erections, 8 (10.3%) were currently responding to pharmacotherapy and 38 (49.4%) failed. The maintenance flow was 75 +/- 45 ml. per minute in the group with spontaneous erections and 103 +/- 60 ml. per minute in the failure group (p = 0.20). Mean patient age was 49.8 +/- 11.7 and 49.1 +/- 10.2 years, respectively (p = 0.23). Of the 41 patients wih normal single potential analysis of cavernous electrical activity 28 had full erections postoperatively, 5 responded to pharmacotherapy and 8 failed, compared to 3, 3 and 30, respectively, of the 36 patients with abnormal single potential analysis of cavernous electrical activity. After a mean followup of 21 months (range 6 to 47 months), 4 patients with full erections at 6 months postoperatively currently require intracavernous pharmacotherapy. Our results indicate that single potential analysis of cavernous electrical activity seems to be an important prognostic factor for the postoperative outcome of penile venous surgery for venogenic impotence.

Arterial anatomy and arteriographic diagnosis of arteriogenic impotence

One hundred twenty-six bilateral selective arteriographic examinations of the iliopudendal vascular tree were performed after comprehensive multidisciplinary evaluation in patients with chronic erectile dysfunction. Best imaging results were obtained by performing the arteriography under epidural anesthesia after intracavernous injection of a vasoactive drug combination. The arteriography is mandatory prior to revascularization procedures. It is further indicated in primary erectile dysfunction and posttraumatic erectile failure. The importance of cavernosography and selective arteriography in primary erectile dysfunction is stressed. Increasing knowledge about the influence of vasoactive drugs on penile hemodynamics has led to its application in diagnosis and therapy of erectile dysfunction. Pharmacocovernosography, Doppler-ultrasound of penile arteries after intracavernous injection of a vasoactive drug combination, and pharmacoarteriography are refined techniques to prove a vascular etiology of erectile dysfunction. The results of the morphologic studies of the vascular system are correlated with functional testing of erectile capacity by intracavernous application of a papaverinephentolamine drug combination.

Transcutaneous Registration of Cavernous Smooth Muscle Electrical Activity: Noninvasive Diagnosis of Neurogenic Autonomic Impotence

Registration of cavernous electrical activity was shown to be a possible method for the evaluation of cavernous autonomic innervation. Recent studies in patients with normal erectile function showed that cavernous electrical activity is synchronous throughout the entire cavernous bodies. Therefore, we examined the feasibility of transcutaneous registration of cavernous electrical activity in 8 normal and 62 impotent patients. In the sitting patient cavernous electrical activity was recorded with a 2-channel electrophysiological unit. Recording was done with a coaxial needle electrode in the proximal left cavernous body and with surface electrodes bilaterally on the penile shaft. In 7 of 8 normal patients swelling of the penile shaft after circumcision resulted in a dramatically decreased amplitude of the potentials. In 41 of 62 impotent patients recordings were similar. In 10 of 62 patients no recording or markedly decreased amplitudes were noted with the surface electrodes and in these patients a small penis or penile retraction with consecutive electrode displacement was found. Careful repositioning of the surface electrodes with the patient in the supine position resulted in similar recordings in 9 (inconsistently in 4). In 11 of the 62 patients more information was obtained with the surface than with the needle electrode. Our results show that recording of cavernous electrical activity can be done in a completely noninvasive manner using surface electrodes with similar or even better information obtained than with needle electrodes.

Diagnostic accuracy of Doppler ultrasound technique of the penile arteries in correlation to selective arteriography

In 63% of 265 patients with erectile dysfunction a relevant arterial inflow disturbance was found by Doppler ultrasound examination. Correlation between Doppler and arteriography in 58 patients showed an accuracy of 95% in detecting penile arteries and an accuracy of 91% in discovering a pathological arterial pattern (arterial anomaly or arteriosclerotic obstruction). In 15 patients the arterial inflow was measured additionally by Doppler ultrasound technique after intracavernosal injection of vasoactive drugs (IIVD) (7.5 mg papaverine and 0.25 mg phentolamine). This technique proved to be more reliable than in the flaccid state and markedly facilitated localization and assessment of pathological changes of the cavernosal arteries.

Exposure of human seminal vesicle tissue to phosphodiesterase (PDE) inhibitors antagonizes the contraction induced by norepinephrine and increases production of cyclic nucleotides.

OBJECTIVES To investigate further the role of phosphodiesterase (PDE) isoenzymes in the control of human seminal vesicle (SV) smooth muscle contractility, we examined the functional responses of isolated SV tissue to various PDE inhibitors. It has been suggested that the application of inhibitors of the PDE type 5 may facilitate SV smooth muscle relaxation and, subsequently, retard ejaculatory response. METHODS Using the organ bath technique, strip preparations of human SV were exposed for 5 minutes to 1 μM of the PDE inhibitors milrinone (PDE3 inhibitor), rolipram, Ro 20-1724 (PDE4 inhibitors), and sildenafil (PDE5 inhibitor). Norepinephrine (NE, alpha agonist) was then added (0,1 μM, 1 μM, and 10 μM) and isometric responses were recorded. A contraction-response curve to NE in the absence of PDE inhibitors was also generated. Drug effects on the production of cyclic adenosine monophosphate (AMP) and cyclic guanosine monophosphate (GMP) were measured by means of radioimmunometric assays. RESULTS The contraction induced by NE was effectively antagonized by 1 μM of rolipram (83.3% inhibition), Ro 20-1724 (72.3% inhibition), sildenafil (41.6% inhibition), and milrinone (37.5% inhibition). The inhibition of force generation was paralleled by a 1.6-fold to 2.8-fold increase in tissue cyclic AMP (induced by milrinone, rolipram, Ro 20-1724), and a 12-fold rise in cyclic GMP (induced by sildenafil). CONCLUSION The findings demonstrate that PDE inhibitors can counteract the contraction of human SV mediated by alpha-adrenergic receptors and enhance levels of cyclic nucleotides. This might be of importance with regard to the identification of new options for the pharmacological treatment of premature ejaculation.

Benign testicular tumors: a case for testis preservation?

Between June 1980 and June 1986, 345 inguinal explorations for intrascrotal space-occupying lesions were performed. In 47 patients benign testicular tumors were diagnosed (13.5%). In 32 of the 47 tumors (68%), the testis could be preserved by excision or enucleation of the tumor. In the remaining 15 patients a high semicastration was carried out: in 12 cases macroscopic examination or frozen section revealed doubtful benignity (6 Leydig cells tumors, 5 dermoid cysts and 1 epidermoid cyst), in 3 patients because of extensive involvement of the testis. In a follow-up period of 1-7 years none of the patients has shown evidence of tumor recurrence or metastasis. Changing the concept of not incising the parietal tunica of the testis and regarding clear-cut macroscopic and histological criteria of benignity on frozen section will safely lower the testicular loss rate from up to 80% to about 30% when benign tumors are encountered.

Characterization of the effects of various drugs likely to affect smooth muscle tension on isolated human seminal vesicle tissue.

OBJECTIVES To investigate the effects of different classes of drugs on the isometric tension of isolated human seminal vesicle (SV) tissue. The contractility of human SV contributes to the process of seminal emission during ejaculation. Different endogenous compounds, such as serotonin (5-HT), adenosine triphosphate (ATP), and nitric oxide, have been suggested to be involved in the control of contraction and relaxation of human SV smooth muscle. However, only limited data are available regarding the effects of compounds known to affect smooth musculature on SV contractile activity. METHODS Using the organ bath technique, the effects of increasing concentrations (10 nm-1 microm/10 microm) of norepinephrine (NE), phenylephrine, endothelin 1, ATP, and 5-HT on human SV tissue at basal tension were studied. In another set-up, SV strip preparations were preincubated with prazosin (alpha-adrenergic blocker), nifedipine and verapamil (Ca(2+)-channel blockers), 2-aminoethoxydiphenyl borate [inositol 1,4,5-trisphosphate (IP(3)) antagonist], cromakalim (K(+)-channel opener), or Y-27632 (ROK inhibitor) (1 microm each, for 10 minutes), followed by the application of NE (0.1 microM, 1 microM, and 10 microm). RESULTS SV smooth muscle was most effectively contracted by NE (mean = 75% of calibrated scale), phenylephrine (mean = 82% of calibrated scale), and endothelin 1 (mean = 70% calibrated scale), whereas only minor responses to ATP (mean = 10.65% calibrated scale) and 5-HT (mean = 6.3% calibrated scale) were observed. The contraction induced by NE was significantly inhibited after pre-exposure of the tissue to prazosin (-92.4%), cromakalim (-83.7%), 2-aminoethoxydiphenyl borate (-43.1%), Y-27632 (-42.8%), and nifedipine (-32.7%). CONCLUSIONS alpha-adrenoceptor antagonism, activation of potassium channels, and inhibition of Rho-kinase decrease the sympathetic contraction of SV smooth muscle. This might be of significance with regard to the identification of new pharmacologic avenues to affect the male ejaculatory system.

THE INFLUENCE OF ANTERIOR ROOT STIMULATION (S2) IN DEAFFERENTED SPINAL CORD INJURY MEN ON CAVERNOUS ELECTRICAL ACTIVITY

Registration of cavernous electrical activity (single potential analysis of cavernous electrical activity) was recently suggested for the diagnosis of autonomic cavernous dysfunction. For validation of this method the effect of sacral neurostimulation on cavernous electrical activity was examined. In 3 male patients with a complete spinal cord lesion (T3, T4 and T12, respectively), deafferentation was done at the S2 to S5 levels. Around the anterior roots of S2 to S5 electrodes for neurostimulation were placed. Cavernous electrical activity was recorded with an intracavernous needle electrode and with surface electrodes in the flaccid as well as in the erect states induced by neurostimulation (at 7, 8, 12, 18, 20, 30 and 45 Hz., 30 v. and 0.4 msec). In all patients similar potentials compared to the normal values, as well as additional pathological potentials were recorded during flaccidity. During neurostimulation all patients achieved full erection with no or markedly decreased cavernous electrical activity to frequencies of 12, 18, 20 and 30 Hz., while to 7, 8 and, more pronounced, 45 Hz. only partial erection with ongoing cavernous electrical activity was found. Our study strongly suggests that cavernous electrical activity and, subsequently, the cavernous smooth muscle tone are dependent on autonomic input. This finding supports the hypothesis that single potential analysis of cavernous electrical activity may be valid in the diagnosis of cavernous autonomic dysfunction. Furthermore, our results suggest a possible role for single potential analysis of cavernous electrical activity in the fine tuning of erection inducing neurostimulators.

Single potential analysis of cavernous electrical activity

SummaryRecording of cavernous electric activity was performed in 178 patients with erectile dysfunction and in 37 normal patients. In 34/37 normal patients, potentials of a uniform shape were recorded during flaccidity: At cut-off frequences of 0.5–500 Hz, the length was 8–18 (mean 12.8, SD 2.8), the amplitude 250–750 (mean 444, SD 109) μV, and the polyphasity 8–22 (mean 13.8, SD 3.3). In impotent patients with upper motor neuron lesions or peripheral lesions, specific types of potentials were observed. In 11/14 impotent patients with insulin-dependent diabetes for over 20 years and clinical findings of cavernous myopathy, potentials showed low amplitude, irregular shape, and slow depolarizations. In 51% of the consecutive impotent patients, abnormal findings of cavernous electric activity were recorded. Our clinical study suggests that single potential analysis of cavernous electric activity (SPACE) may be useful in the diagnosis of cavernous autonomic neuropathy and cavernous smooth muscle myopathy.