Pelvic Nerve Stimulation Research Articles

Spinal neurons activated in response to pudendal or pelvic nerve stimulation in female rats

Spinal neurons activated in response to pudendal or pelvic nerve stimulation in female rats

ORIGINAL RESEARCH–BASIC SCIENCE: Expression of Aquaporin Water Channels in Rat Vagina: Potential Role in Vaginal Lubrication

Aquaporins (AQPs) are membrane proteins that facilitate water movement across biological membranes. There has been little research on the role of AQPs in the female sexual arousal response. The purposes of this study were to investigate the localization and functional roles of AQP1, AQP2, and AQP3 in rat vagina. Female Sprague-Dawley rats (230-240 g, N = 20) were anesthetized. The vaginal branch of the pelvic nerve was stimulated for 60 seconds (10 V, 16 Hz, 0.8 ms), and the animals were sacrificed either immediately or 5 minutes later. The expression and cellular localization of AQP1, 2, and 3 were determined by Western blot and immunohistochemistry of the vagina. The intracellular membrane and plasma membrane fractions of the proteins in vaginal tissue were studied by immunoblot analysis with the differential centrifugation. The expression and cellular localization of AQPs, and pelvic nerve stimulation induced translocation of AQPs in rat vaginal tissue. Immunolabeling showed that AQP1 was mainly expressed in the capillaries and venules of the vagina. AQP2 was expressed in the cytoplasm of the epithelium, and AQP3 was mainly associated with the plasma membrane of the vaginal epithelium. AQPs were found to be present primarily in the cytosolic fraction of untreated tissues. The translocation of AQP1 and 2 isoforms from the cytosolic compartment to the membrane compartment was observed immediately after nerve stimulation and had declined at 5 minutes after nerve stimulation, while the subcellular localization of AQP3 was not changed by nerve stimulation. These results showed a distinct localization of AQPs in the rat vagina. Pelvic nerve stimulation modulated short-term translocation of AQP1 and 2. These results imply that AQPs may play an important role in vaginal lubrication.

Pharmacological effects of propiverine and its active metabolite, M‐1, on isolated human urinary bladder smooth muscle, and on bladder contraction in rats

To investigate the effects of M-1, a major active metabolite of propiverine on the bladder. We have evaluated the effects of M-1 on the contractions induced by carbachol, KCl, CaCl(2), and electrical field stimulation (EFS) in human detrusor smooth muscles, and pelvic nerve stimulation-induced bladder contractions in rats. The effects of M-1 were also compared with the effects of propiverine and tolterodine. Pretreatment with propiverine and tolterodine caused parallel shifts to the right of the concentration-response curves to carbachol. M-1 caused concentration-dependent reduction in the maximum contractile responses induced by carbachol. Although tolterodine did not inhibit the KCl- and CaCl(2)-induced contractions, M-1 and propiverine significantly inhibited these contractions. In the presence of atropine, M-1 and propiverine significantly inhibited the atropine resistant part of the contraction induced by EFS. On the other hand, tolterodine did not have significant inhibitory effects on atropine resistant contractions. Pelvic nerve stimulation induced bimodal phasic and tonic contractions in the rat bladder. M-1 mainly inhibited the phasic contraction. Tolterodine caused a significant inhibition in the tonic contraction, and propiverine had inhibitory effects on both contractions. The present results suggest that M-1 has inhibitory effects on the bladder smooth muscles through calcium antagonistic action. It is possible that the clinical effects of propiverine on the human bladder are based not only on the action of propiverine itself but also on one of its active metabolites, M-1.

Increased α1-Adrenergic Activity in the Rat Bladder by Depletion of Ovarian Hormones

It is possible that postmenopausal loss of ovarian hormones leads to sensitization and/or over expression of alpha1-adrenergic receptors. The objective of this study was to determine whether ovariectomy enhances efferent transmission and, hence, increases contractility in the rat bladder by increasing alpha1-adrenergic receptors. This hypothesis was tested in 4 urethane anesthetized groups of female rats, including regularly cycling rats in metestrus and proestrus, and rats 4 months after ovariectomy with and without estradiol replacement, respectively. The bladder was catheterized through the urethra to measure intravesical pressure. The left femoral artery was also catheterized for local delivery of the nonspecific alpha1-adrenergic agonist phenylephrine, the alpha1-adrenergic antagonist prazosin, the specific alpha1A antagonist 5-methylurapidil or dimethyl sulfoxide (Sigma) in saline vehicle. In some rats bladder contractility was assessed by cystometry, followed by treatment with phenylephrine injected via the femoral artery. In other rats the bladder was denervated and a platinum electrode was wrapped around branches of the left pelvic nerve for electrical stimulation. The amplitude of micturition contractions and contractions evoked by phenylephrine or pelvic nerve stimulation was measured. Micturition and phenylephrine evoked bladder contractions were significantly increased in ovariectomized rats without estradiol replacement. Prazosin significantly decreased contractions evoked by pelvic stimulation in a dose dependent manner. This decrease was significantly larger in ovariectomized rats without estradiol replacement compared to that in the other groups. The inhibition of stimulation evoked contractions by 5-methylurapidil was also greater in ovariectomized rats without estradiol replacement than in rats with estradiol replacement. Results suggest that the depletion of ovarian hormones after ovariectomy enhances alpha1-adrenergic activity.

The role of free radicals and nitric oxide in the ischemia–reperfusion injury mediated by acute bladder outlet obstruction

Free-radical generation and nitric oxide (NO) generation were detected in the rat bladder following acute bladder outlet obstruction (BOO), and the results were compared with those for vascular ischemia and reperfusion (I-R). Forty male Sprague Dawley rats were used. In the acute BOO plus I-R group (group 1), rats were catheterized with a 3-Fr catheter and an inflated balloon was positioned at the bladder neck. The bladder was overdistended after administration of Ringer solution and furosemide (12 mg/kg, each) for 60 min, and was then drained to allow reperfusion for 30 min. In the acute BOO plus nerve stimulation group (group 2), the pelvic nerve was stimulated in the distended bladder for 60 min (5 s every 5 min, 10 V/0.1 ms, 20 Hz). Pelvic nerve stimulation was performed in nonobstructed animals in group 3. In the I-R group (group 4), the distal aorta was occluded for 60 min followed by reperfusion for 30 min. Sham-operated animals served as the control group (group 5). At the end of the protocols, the levels of hydroxyl and superoxide radicals and NO levels were measured in the bladder tissues with luminol- and lucigenin-enhanced chemiluminescence methods. The results were compared by a one-way analysis of variance test. The levels of hydroxyl radicals were not significantly different between the study groups. In contrast, superoxide radicals and NO levels were significantly increased in both group 1 and group 4 compared with those in control animals (P<0.05 for all comparisons). Superoxide radical generation in group 2 was comparable to the levels in group 1 (P>0.05), whereas NO levels were substantially lower than in group 1 (P=0.06). In summary, vascular I-R causes significant oxidative damage to the bladder. Acute BOO with overdistension of the bladder mimics the effects of true vascular I-R injury. The NO pathway has possibly a major role in I-R-induced bladder damage. Prolonged BOO may therefore significantly enhance the oxidative damage to the bladder and further accentuate the effects of generalized atherosclerotic processes in the elderly adult.

Spinal glutamatergic NMDA-dependent pelvic nerve-to-external urethra sphincter reflex potentiation caused by a mechanical stimulation in anesthetized rats

The current study investigates whether the spinal pelvic nerve-to-external urethra sphincter (EUS) reflex potentiation can be induced by a mechanical stimulation and whether the glutamatergic mechanism is involved in yielding such a reflex potentiation. The external urethra sphincter electromyogram (EUSE) activity, evoked by a single or by repetitive pelvic nerve stimulation, in 30 anesthetized rats was recorded with/without bladder saline distension. Without saline distension (0 cmH(2)O), a single pulse nerve stimulation evoked a single action potential in the reflex activity, whereas repetitive pelvic stimulation and saline distension (6 approximately 20 cmH(2)O) both elicited a long-lasting reflex potentiation (20.05 +/- 3.21 and 75.01 +/- 9.87 spikes/stimulation, respectively). The saline distension-induced pelvic nerve-to-EUS reflex potentiation was abolished by D-2-amino-5-phosphonovalerate [APV; a glutamatergic N -methyl-D-aspartic acid (NMDA) receptor antagonist; 100 microM, 10 microl, 1.72 +/- 0.31 spikes/stimulation] and attenuated by 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F) quinoxaline [NBQX; a glutamatergic alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) receptor antagonist; 100 microM, 10 microl, 26.16 +/- 7.27 spikes/stimulation], but was not affected by bicuculline (a GABAergic antagonist; 100 microM, 10 microl, 53.62 +/- 15.54 spikes/stimulation). Intrathecal administration of glutamate (31.12 +/- 8.25 spikes/stimulation, 100 microM, 10 microl) and NMDA (26.25 +/- 4.12 spikes/stimulation, 100 microM, 10 microl) both induced a long-lasting pelvic nerve-to-EUS reflex potentiation without saline distension, which was similar to the findings observed from saline distension only. The duration of the contraction wave of the urethra was elongated by the saline distension-induced pelvic nerve-to-EUS reflex potentiation, whereas the peak pressure of the contraction wave was not affected. Our findings suggest that saline distension in the bladder elicits a pelvic nerve-to-EUS reflex potentiation and the glutamatergic mechanism contributes to the presence of such a reflex potentiation.

Long-term urinary dysfunction after mesorectal excision: A prospective study with intraoperative electrophysiological confirmation of nerve preservation

Background Bladder dysfunctions are well-recognized complications after nerve-sparing mesorectal excision for rectal cancer. This study sought to symptomatically analyze the extent of recovery from major and minor urinary symptoms in patients with signs of bladder denervation. Methods Sixty-two patients with mesorectal excision for rectal cancer were investigated prospectively. Pelvic autonomic nerve preservation (PANP) was assessed macroscopically and with the aid of intraoperative electrical stimulation of pelvic autonomic nerves (INS). Bladder function was evaluated with the International Prostate Symptom Score (IPSS) and the Quality of life index (Qol). Median follow-up was 20 months (range 3–40 months). Results Forty-six patients with INS-confirmed preservation of parasympathetic nerves remained unchanged in early and long-term urinary function (IPSS: median 1; range 0–24 and Qol 0; range 0–5). In 15 patients without confirmation of PANP (unilaterally or bilaterally) on INS, voiding function was significantly more impaired postoperatively (IPSS: median 10; range 0–25 and Qol 3; range 0–6) and at long-term follow-up (IPSS: median 9; range 0–25 and Qol 3; range 0–6) ( p < 0.001). Voiding function was improved in 4 of 10 patients with major and minor symptoms. In 5 of 6 patients with long-term bladder dysfunction INS assessed parasympathetic nerve damage unilaterally (3/5) and bilaterally (2/5). Conclusion Long-term voiding disturbance after mesorectal excision was found to be a serious complication. INS while monitoring intravesical pressure is a valuable aid in predicting long-term bladder function after TME. The device may serve a secondary preventive function in enabling the initiation of early urologic therapy.

Testosterone Increases Blood Flow and Expression of Androgen and Estrogen Receptors in the Rat Vagina

The mechanisms by which testosterone modulates female genital sexual arousal responses are poorly understood. To investigate the effects of testosterone on vaginal blood flow and the expression of estrogen and androgen receptor proteins in the rat vagina. Mature female Sprague-Dawley rats were sham-operated (intact) or ovariectomized. Fourteen days after ovariectomy, animals were continuously infused with vehicle or varying doses of testosterone (5.5-55 microg/day). After 2 weeks of treatment, vaginal blood flow in response to pelvic nerve stimulation was measured by laser Doppler flowmetry. Plasma levels of testosterone and estradiol were determined by radioimmunoassay and epithelial thickness was examined in fixed vaginal tissue sections. Androgen and estrogen receptor levels were assessed by equilibrium radioligand binding and by Western blot analyses. Vaginal blood flow responses were significantly reduced in ovariectomized rats and normalized in animals infused with testosterone. Ovariectomy increased the expression of estrogen receptors and reduced the expression of androgen receptors with no change in receptor-ligand affinity. Testosterone increased the expression of both androgen and estrogen receptors in the vagina. While physiological (11 microg/day) and supraphysiological (55 microg/day) concentrations of testosterone normalized vaginal tissue weight, uterine tissue and whole body weights were not significantly different from ovariectomized rats infused with vehicle. Testosterone infusion, even at supraphysiological concentrations, did not change plasma estradiol levels when compared to vehicle-infused, ovariectomized rats. Likewise, the vaginal epithelium of testosterone-infused rats remained atrophic, similar to vehicle-infused, ovariectomized rats, indicating that testosterone is not aromatized to estrogens at significant levels in the vagina. Our data suggest that testosterone regulates androgen and estrogen receptor protein expression in the vagina and enhances vaginal perfusion by an androgen-dependent mechanism. We conclude that testosterone plays an important role in modulating the physiology of the vagina and contributes to improvement of genital sexual arousal responses.

Intraoperative electrostimulation objectifies the assessment of functional nerve preservation after mesorectal excision

To improve nerve-sparing surgery, intraoperative electrical stimulation of pelvic autonomic nerves (INS) has been proposed in urology, gynecology, and visceral surgery. The aim of this study was to assess the impact of INS while monitoring intravesical pressure on the accurate evaluation of pelvic autonomic nerve preservation (PANP) after mesorectal excision. It was sought to determine whether this confirmation is useful in the prediction of postoperative urinary function. Sixty-two patients with mesorectal exzision for rectal cancer were examined prospectively. PANP was assessed visually by the surgeon and with INS. Bladder function was evaluated by post voiding residual volume measurement, rate of recatheterization, rate of long-term urinary catheterisation, and the international prostatic symptom score with quality of life index. INS confirmed bilateral preservation of parasympathetic nerves in 46 patients (74%), and in 10 patients (16%) in at least one side. In six patients (10%), INS failed to confirm PANP. Eleven patients (18%) developed urinary symptoms postoperatively. INS results had a higher sensitivity than visual assessment by the surgeon (82 vs 46%). Values for specificity ranged at 90 and 92%, respectively. Accuracy of INS in predicting PANP was higher (88 vs 83%). The correlation between urinary function and the findings on INS was good (kappa-value: 0.65), correlation between urinary function and visual assessment by the surgeon was fair (kappa-value: 0.40). INS, while monitoring intravesical pressure, accurately predicts bladder function after mesorectal excision. It may provide further insight into pelvic autonomic nerve sparing techniques.

Effects of tamoxifen on vaginal blood flow and epithelial morphology in the rat

BackgroundTamoxifen, a selective estrogen receptor modulator with both estrogenic and anti-estrogenic activity, is widely used as adjuvant therapy in breast cancer patients. Treatment with tamoxifen is associated with sexual side effects, such as increased vaginal dryness and pain/discomfort during sexual activity. There have been limited investigations of the effect of tamoxifen on estrogen-dependent peripheral genital arousal responses. The objective of this study was to investigate the effects of tamoxifen on vaginal physiology in the rat.MethodsFemale Sprague-Dawley rats were subjected to sham surgery or bilateral ovariectomy. After 2 weeks, sham-operated rats were implanted with subcutaneous osmotic infusion pumps containing vehicle (control) or tamoxifen (150 μg/day). Ovariectomized rats were similarly infused with vehicle. After an additional 2 weeks, vaginal blood flow responses to pelvic nerve stimulation were measured by laser Doppler flowmetry and vaginal tissue was collected for histological and biochemical assay.ResultsTamoxifen treatment did not change plasma estradiol concentrations relative to control animals, while ovariectomized rats exhibited a 60% decrease in plasma estradiol. Tamoxifen treatment caused a significant decrease in mean uterine weight, but did not alter mean vaginal weight. Vaginal blood flow was significantly decreased in tamoxifen-infused rats compared to controls. Similar to ovariectomized animals, estrogen receptor binding was increased and arginase enzyme activity was decreased in tamoxifen-infused rats. However, different from control and ovariectomized animals, the vaginal epithelium in tamoxifen-infused rats appeared highly mucified. Periodic acid-Schiff staining confirmed a greater production of carbohydrate-rich compounds (e.g. mucin, glycogen) by the vaginal epithelium of tamoxifen-infused rats.ConclusionThe observations suggest that tamoxifen exerts both anti-estrogenic and pro-estrogenic effects in the vagina. These physiological alterations may eventually lead to vaginal atrophy and compromise sexual function.

Recurrent inhibition of the bladder C fibre reflex in the cat and its response to naloxone

Recurrent inhibition of the bladder C fibre reflex was studied in adult female cats anaesthetized with alpha-chloralose. Test reflexes were evoked by electrical stimulation of bladder Adelta and C afferents in the right pelvic nerve and were recorded from the proximal end of a small ipsilateral pelvic nerve branch, transected close to the bladder. Such test reflexes were consistently depressed by repetitive electrical stimulation of the contralateral bladder pelvic nerve (20 Hz, 20 s) at intensities sufficient to recruit axons of bladder preganglionic neurones. The inhibition could be evoked after transection of the left dorsal roots S1-S4 and the sympathetic supply to the bladder but was abolished by transection of the pelvic nerve central to the site of stimulation. Hence, it most likely involved central recurrent collaterals of antidromically activated bladder preganglionic neurones. The reflex suppression was quite considerable - maximal C fibre reflexes were reduced to a group mean of 25% (+/- 9% confidence interval) of their control size. The effect had a slow onset, requiring a few seconds of conditioning stimulation to be revealed, and was very long lasting (minutes). Naloxone (0.01-0.5 mg kg(-1) i.v.) abolished the recurrent inhibition of both the C fibre and Adelta bladder reflexes, while inhibition from afferents in the dorsal clitoris nerve remained unchanged. It is concluded that the segmental bladder C fibre reflex and the spino-ponto-spinal Adelta micturition reflex are both targets of recurrent inhibition from bladder parasympathetic preganglionic neurones and that the effect involves an enkephalinergic mechanism.

Ano‐rectal motility responses to pelvic, hypogastric and pudendal nerve stimulation in the Göttingen minipig

We investigated the effect of efferent stimulation of the pelvic (PN), hypogastric (HGN) and pudendal (PuN) nerves on ano-rectal motility in Göttingen minipigs using an impedance planimetry probe. Changes in the rectal cross-sectional area (CSA) at five axial positions and pressures in the rectum and anal canal were investigated simultaneously. Pelvic nerve stimulation elicited a CSA decrease in the proximal part of the rectum and a simultaneous CSA increase in its distal part. Anal pressure also decreased. Hypogastric nerve and PuN stimulation elicited an increase in anal pressure, but no rectal response. Severing the HGN produced a persistent reduction in resting anal pressure, but no change was observed when the PN and the PuN were severed. Stimulation of the distal part of all three nerves produced a persistent response. Administration of phentolamine and pancouronium eliminated the response to stimulation of the HGN and the PuN, respectively. Rectal responses to PN stimulation vary more than previously suggested. The HGN has an excitatory effect on the internal anal sphincter, and the PuN on the external anal sphincter. However, the PuN plays no major role in maintaining basal anal pressure.

Effects of Korean Red Ginseng on the Vaginal Blood Flow and Structure in Female Castrated Rats

Purpose: Ginseng is a traditional Asian remedy for sexual dysfunction. The purposes of this study were to investigate the effects of Korean red ginseng (KRG) on the vaginal blood flow and tissue structure in female castrated rats. Materials and Methods: Female Spague-Dawley rats (200-210gm) were divided into 4 groups: the control (n=20), castration (n=30), and castration plus oral administration of KRG extracts (50 and 100mg/kg/day, n=15 and n=15, respectively). After 1 month of treatment, the serum estrogen and total cholesterol levels were measured. The vaginal blood flow was measured using laser Doppler flowmeter before and after pelvic nerve stimulation (PNS). The vaginal tissue was processed for Masson's trichrome stain, immunohistochemistry and Western blotting. Results: The serum estrogen level was significantly decreased in the castration group (0.8±1.9ng/ml); however, it increased up to the control level (2.2±1.3ng/ml) in both the KRG administration groups (p<0.05). The PNS-induced vaginal blood flow tended to improve in the KRG treatment groups. On the histology, the vaginal epithelial layer and submucosal microvasculature showed improvement in the KRG treatment groups. The expression of estrogen receptor increased in the KRG treatment groups compared to the castration group. Conclusions: These results suggested that KRG extracts seem to have an estrogenic effect on castrated female rats. This implies that the KRG extracts may have an ameliorating effect on sexual function in menopausal woman. (Korean J Urol 2006;47:888-894) ꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏ

Streptozotocin-induced diabetes in the rat is associated with changes in vaginal hemodynamics, morphology and biochemical markers

BackgroundDiabetes is associated with declining sexual function in women. However, the effects of diabetes on genital tissue structure, innervation and function remains poorly characterized. In control and streptozotocin-treated female rats, we investigated the effects of diabetes on vaginal blood flow, tissue morphology, and expression of arginase I, endothelial nitric oxide synthase (eNOS) and cGMP-dependent protein kinase (PKG), key enzymes that regulate smooth muscle relaxation. We further related these changes with estrogen receptor alpha (ERα) and androgen receptor (AR) expression.ResultsIn addition to significantly elevated blood glucose levels, diabetic rats had decreased mean body weight, lower levels of plasma estradiol, and higher plasma testosterone concentration, compared to age-matched controls. Eight weeks after administration of buffer (control) or 65 mg/kg of streptozotocin (diabetic), the vaginal blood flow response to pelvic nerve stimulation was significantly reduced in diabetic rats. Histological examination of vaginal tissue from diabetic animals showed reduced epithelial thickness and atrophy of the muscularis layer. Diabetic animals also had reduced vaginal levels of eNOS and arginase I, but elevated levels of PKG, as assessed by Western blot analyses. These alterations were accompanied by a reduction in both ERα and AR in nuclear extracts of vaginal tissue from diabetic animals.ConclusionIn ovariectomized (estrogen deficient) animals, previous reports from our lab and others have documented changes in blood flow, tissue structure, ERα, arginase I and eNOS that parallel those observed in diabetic rats. We hypothesize that diabetes may lead to multiple disruptions in sex steroid hormone synthesis, metabolism and action. These pathological events may cause dramatic changes in tissue structure and key enzymes that regulate cell growth and smooth muscle contractility, ultimately affecting the genital response during sexual arousal.

Intraoperative electrical stimulation of the pelvic splanchnic nerves during nerve-sparing radical hysterectomy

Objectives. This study sought to determine whether intraoperative electrical stimulation (IES) of pelvic splanchnic nerves (PSNs) while monitoring bladder contraction was useful to predict postoperative bladder function during conventional nerve-sparing radical hysterectomy. Methods. Seventeen patients with stage Ib or IIa cervical cancer underwent conventional radical hysterectomy. IES was performed in all cases, stimulating the roots of PSN, the posterior sheath of the vesicouterine ligament (PVL) and the dorsal area of the ligament. After resection of the uterus, the PSN roots were stimulated again. Bladder function was evaluated by urodynamic study (UDS) preoperatively and 3 months after surgery. Results. The results of IES were consistent with bladder function evaluated by postoperative UDS. In 13 of 17 cases, an increased intravesical pressure was observed with IES of the PSN roots after uterus resection. Nine of 13 cases showed marked detrusor contraction with UDS 3 months after surgery and were able to void without using abdominal pressure except in one case. In the remaining 4 of 17 cases, no response could be detected to IES on either side. Three cases voided using abdominal pressure and one used clean intermittent self-catheterization without spontaneous voiding. Conclusions. IES while monitoring intravesical pressure during radical hysterectomy represents a technically simple and useful procedure for the prediction of postoperative bladder function.

Adipocyte Accumulation in Penile Corpus Cavernosum of the Orchiectomized Rabbit: A Potential Mechanism for Veno‐occlusive Dysfunction in Androgen Deficiency

Androgens are deemed to be critical for the development, growth, and maintenance of penile tissue as well as for erectile function. Androgens are also reported to inhibit differentiation of stroma progenitor cells into adipocytes and promote differentiation into smooth muscle. The objective of this study was to investigate whether androgen deprivation results in accumulation of adipocytes in the corpus cavernosum. Mature, New Zealand white male rabbits were subjected to sham surgery (control) or orchiectomy. Two weeks after surgery, erectile function was assessed by monitoring changes in intracavernosal blood pressure (ICP) in response to pelvic nerve stimulation. All ICP measurements were normalized to the mean systemic arterial blood pressure. In parallel studies, penile cross sections from control and orchiectomized rabbits were fixed and stained with either Masson's trichrome or hematoxylin and eosin to assess smooth muscle and connective tissue content. Alternatively, tissue sections were stained with Toluidine blue to assess accumulation of fat-containing cells. Orchiectomy resulted in loss of erectile function and penile atrophy, associated with reduced trabecular smooth muscle and increased connective tissue content. Most strikingly, tissue from orchiectomized animals exhibited accumulation of fat-containing cells (adipocytes) in the subtunical region of the corpus cavernosum. We hypothesize that androgen deprivation promotes differentiation of progenitor stroma cells into an adipogenic lineage producing fat-containing cells, thus altering erectile function.

Validity of Pelvic Autonomic Nerve Stimulation With Intraoperative Monitoring of Bladder Function Following Total Mesorectal Excision for Rectal Cancer

This prospective study was designed to clarify whether the results of the intraoperative stimulation of parasympathetic pelvic nerves performed in 31 patients after mesorectal excision for rectal carcinoma allowed predictions in terms of the postoperative bladder function of the patients. After monopolar stimulation of the splanchnic pelvic nerves using a constant voltage stimulator (Screener 3625), intravesical pressure increase was measured manometrically. The results were related to the postoperative residual urine volume, requirement of recatheterization and long-term catheterization, just as to the results of the validated International Prostatic Symptom Scores and the Quality of Life Index caused by urinary symptoms. The median follow-up period was nine (range, 2-14) months. Parasympathetic nerve stimulation was performed at 61 sites and results in intravesical pressure increase up to 6 cm water column in median. In 11 patients (33.3 percent), a negative test result was achieved: 5 with unilateral and 6 with bilateral pressure increases of < or = 2 cm water column. Recatheterization was necessary in four patients, and all of them showed negative neuromonitoring results. Two of these patients were discharged with an in situ urinary bladder catheter. Postoperative increased residual urine volumes (> or =100 ml) resulted more frequently in the group with negative test results (63.6 vs. 21.1 percent; P = 0.047), and the International Prostatic Symptom Score and Quality of Life Index showed the worst results (9.9 +/- 6.7 vs. 3 +/- 4.9, P = 0.021; 2.4 +/- 1.7 vs. 0.7 +/- 1.3, P = 0.021). Intraoperative neurostimulation and manometric measurement of bladder pressure may contribute to the identification of parasympathetic pelvic nerves during total mesorectal excision. This method is suitable for intraoperative recording of nerve preservation and therefore associated with postoperative bladder function.

MODULATION OF RAT VAGINAL BLOOD FLOW AND ESTROGEN RECEPTOR BY ESTRADIOL

The effects of subphysiological and physiological levels of estradiol on vaginal blood flow and estrogen receptor were investigated. Intact or ovariectomized female Sprague-Dawley rats were used. Two weeks after surgery rats were infused with vehicle (polyethyleneglycol), or estradiol at subphysiological (5 microg daily) or physiological (15 microg daily) concentrations for 14 days using osmotic pumps. Changes in vaginal blood flow elicited by pelvic nerve stimulation were assessed by laser Doppler flowmetry. Total levels of functional estrogen receptor were determined by radioligand binding and Western blot analyses were used to assess estrogen receptor (ER) alpha protein. Mean plasma estradiol concentration +/- SEM decreased by 63% in the vehicle group (intact 36.5 +/- 10.3 pg/ml). The subphysiological and physiological estradiol groups had plasma levels that were 55% and 83% of the intact group, respectively. Uterine and vaginal wet weight, and vaginal blood flow were significantly decreased in the vehicle group and normalized by physiological levels of estradiol. However, vaginal blood flow was significantly greater in the subphysiological estradiol group compared to intact animals. Specific binding of [H]estradiol in vaginal tissue extracts from intact rats was 0.51 fmol/mg protein and it was increased 30-fold in the vehicle group. ER binding in vaginal tissue in the physiological estradiol group decreased to levels that were comparable to those in intact animals, whereas estrogen receptor binding remained elevated in the subphysiological estradiol group. These changes were paralleled by ERalpha protein levels. Estradiol is crucial for maintaining optimal vaginal blood flow in the rat. Lower levels of plasma estradiol trigger compensatory ERalpha up-regulation.

Role of adenosine triphosphate-dependent potassium channels in canine penile erection

Objectives To define the physiologic role and hemodynamic features of nitric oxide (NO) and the adenosine triphosphate (ATP)-dependent K + (K ATP) channel in canine penile erection. Methods Mongrel dogs were anesthetized, and penile erection was induced by electrical stimulation of the pelvic nerve. Changes in the intracavernous pressure (ICP) were measured with a transducer. Results The basal ICP was 12.8 ± 5.0 mm Hg. Pelvic nerve stimulation (5 to 20 V, 5 to 15 Hz, for 1-minute intervals) significantly increased the ICP to 86.2 ± 11.4 mm Hg (n = 5, P <0.05). Treatment with the NO synthase inhibitor N ω-nitro- l-arginine methyl ester (10 mg/kg intravenously) abolished this increase (15.4 ± 5.0 mm Hg, n = 5). Intracavernous injection of the K ATP channel opener cromakalim (3 and 10 μg) increased the ICP (103 ± 14.4 mm Hg and 106 ± 12.1 mm Hg, respectively; n = 4). This response was abolished by the prior intracavernous injection of the selective K ATP channel-specific blocker glibenclamide (10 mg). Glibenclamide did not affect the increase in ICP induced by electric stimulation of the pelvic nerve (88 ± 24.2 mm Hg). Conclusions Our results indicate that relaxation of canine cavernous smooth muscle and penile tumescence are mediated by NO. The failure of glibenclamide to affect the increase in ICP induced by pelvic nerve stimulation suggests that ATP-dependent K + channels probably do not play a physiologic role in canine penile erection.

Brain stem convergence of pelvic viscerosomatic inputs via spinal and vagal afferents.

Single medullary reticular formation (MRF) neurons receive ascending spinal inputs from multiple somatic and pelvic visceral territories. MRF neurons were examined for responses to both pelvic (PN) and vagus (abdominal branches: VAG-abd) nerve stimulation, which dually innervate certain pelvic viscera. Recordings in 12 urethane-anesthetized male rats were performed. Of 121 PN-responsive MRF neurons, 50% responded to VAG-abd. Twenty-seven (22%) responded to colonic distention. All 121 neurons responded to noxious stimulation of somatic territories, including many areas outside the perigenital region (including the hindpaws, ears, face). These data demonstrate input originating from different spinal and cranial nerves. The functional significance of this viscerosomatic convergence to MRF is unknown, but could relate to sensory/autonomic integration for coordinating multiple bodily functions, including reproductive and eliminative events.