Paul Zaszczurynski

Striated Muscle and Nerve Fascicle Distribution in the Female Rat Urethral Sphincter

The anatomical basis for urinary continence depends on a thorough understanding of the tissues in the urethra. The objective of this study was to evaluate the morphology and neuroanatomy of urethral striated muscle, called the rhabdosphincter or external urethral sphincter, in normal female rats. Urethras from 12 female rats were dissected from the bladder, fixed, embedded in paraffin or epon, and sectioned every 1 mm. Striated muscle content was taken as the ratio of the striated muscle area to net urethral area. Nerve fascicles containing myelinated axons near the rhabdosphincter were counted and mapped. Both striated muscle content and number of nerve fascicles peak in the proximal third of the urethra, with a secondary peak at the distal end of the urethra. This secondary peak may correspond to an analog of the combined compressor urethrae/urethrovaginal sphincter located in the distal urethra in human. The rhabdosphincter has a variable distribution along the length of the urethra. In the middle and distal thirds of the urethra, the dorsal striated muscle fibers between the urethra and vagina become more sparse. The majority of nerve fascicles are contained in the lateral quadrants of the urethra, similar to the lateral distribution of somatic nerves in humans. In conclusion, this study demonstrates the normal distribution of the striated musculature and neuroanatomy in the urethra, with similarities to the human. It thus supports and extends the usefulness of the rat as an experimental model for studying urinary incontinence. Anat Rec 290:145–154, 2007.

Dual simulated childbirth injury delays anatomic recovery

A dual childbirth injury model, including vaginal distension (VD) and pudendal nerve crush (PNC), may best represent the injuries seen clinically. The objective of this study was to investigate urethral function, anatomy, and neurotrophin expression after several simulated childbirth injuries. Groups of 140 rats underwent PNC, VD, PNC+VD, or neither (C). Four days after injury, all injury groups had significantly decreased leak-point pressure (LPP) compared with C rats. Ten days after injury, LPP in PNC and PNC+VD rats remained significantly lower than C rats. Three weeks after injury, LPP in all injury groups had recovered to C values. Histological evidence of injury was still evident in the external urethral sphincter (EUS) after VD and PNC+VD 10 days after injury. Three weeks after injury, the EUS of PNC+VD rats remained disrupted. One day after VD, brain-derived neurotrophic factor (BDNF) expression in the EUS was reduced, while neurotrophin-4 (NT-4) and nerve growth factor (NGF) expression was unchanged. BDNF, NT-4, and NGF expression was dramatically upregulated in the EUS after PNC. After PNC+VD, NGF expression was upregulated, and BDNF and NT-4 expression was upregulated somewhat but not to the same extent as after PNC. Ten days after injury, PNC+VD had the least number of normal nerve fascicles near the EUS, followed by PNC and VD. Twenty-one days after injury, all injury groups had fewer normal nerve fascicles, but without significant differences compared with C rats. PNC+VD therefore provides a more severe injury than PNC or VD alone.

Effects of sphincterotomy and pudendal nerve transection on the anal sphincter in a rat model

PURPOSE: Our objective was to define anal resting pressure and electromyography of the normal rat anal sphincter and investigate the short-term effects of both mechanical trauma to the anal sphincter muscles and pudendal nerve transection. METHODS: Forty-five virgin female Sprague-Dawley rats were randomly allotted to three groups: controls (n = 21), sphincterotomy (n = 12), and pudendal nerve transection (n = 12). Anal pressure was monitored using a saline-filled balloon connected to a pressure transducer. Anal pressure and electromyography of the anal sphincter with use of a needle electrode were recorded both before and after injury or succinylcholine administration. RESULTS: Anal pressure data were consistent with rhythmic pressure contractions. Succinylcholine significantly reduced both pressure and electromyography signals. Electromyography amplitude and frequency decreased after nerve transection but not after sphincterotomy. The histology showed that the rat anal anatomy has muscular components that compare with human anatomy. The sphincterotomy group showed injury to the anal sphincters and the sphincter anatomy of the nerve transection group appeared similar to the control group. The anal pressure wave appears to be created by synergistic activity of both striated and smooth muscle of the anal sphincter. CONCLUSION: The female rat is a suitable and reliable model for studying effect of direct and indirect injury to the anal sphincters.

Quantitative evaluation of electrodes for external urethral sphincter electromyography during bladder-to-urethral guarding reflex

PurposeAccuracy in the recording of external urethral sphincter (EUS) electromyography (EMG) is an important goal in the quantitative evaluation of urethral function. The aim of this study was to quantitatively compare electrode recordings taken during tonic activity and leak point pressure (LPP) testing.MethodsSeveral electrodes, including the surface electrode (SE), concentric electrode (CE), and wire electrode (WE), were placed on the EUS singly and simultaneously in six female Sprague-Dawley rats under urethane anesthesia. The bladder was filled via a retropubic catheter while LPP testing and EUS EMG recording were done. Quantitative baseline correction of the EUS EMG signal was performed to reduce baseline variation. Amplitude and frequency of 1-s samples of the EUS EMG signal were measured before LPP (tonic activity) and during peak LPP activity.ResultsThe SE, CE, and WE signals demonstrated tonic activity before LPP and an increase in activity during LPP, suggesting that the electrodes accurately recorded EUS activity during tonic activity and during the bladder-to-EUS guarding reflex, regardless of the size or location of detection areas. SE recordings required significantly less baseline correction than both CE and WE recordings. The activity in CE-recorded EMG was significantly higher than that of the SE and WE both in single and simultaneous recordings.ConclusionsThese electrodes may be suitable for testing EUS EMG activity. The SE signal had significantly less baseline variation and the CE detected local activity more sensitively than the other electrodes, which may provide insight into choosing an appropriate electrode for EUS EMG recording.

Somatomotor and sensory urethral control of micturition in female rats

In rats, axons of external urethral sphincter (EUS) motoneurons travel through the anastomotic branch of the pudendal nerve (ABPD) and anastomotic branch of the lumbosacral trunk (ABLT) and converge in the motor branch of the sacral plexus (MBSP). The aim of the present study was to determine in female rats the contribution of these somatomotor pathways and urethral sensory innervation from the dorsal nerve of the clitoris on urinary continence and voiding. EUS electromyographic (EMG) activity during cystometry, leak point pressure (LPP), and voiding efficiency (VE) were assessed in anesthetized virgin Sprague-Dawley female rats before and after transection of the above nerve branches. Transection of the MBSP eliminated EUS EMG, decreased LPP by 50%, and significantly reduced bladder contraction duration, peak pressure, intercontraction interval, and VE. Transection of the ABPD or ABLT decreased EUS EMG discharge and LPP by 25% but did not affect VE. Transection of the dorsal nerve of the clitoris did not affect LPP but reduced contraction duration, peak pressure, intercontraction interval, and VE. We conclude that somatomotor control of micturition is provided by the MBSP with axons travelling through the ABPD and ABLT. Partial somatomotor urethral denervation induces mild urinary incontinence, whereas partial afferent denervation induces voiding dysfunction. ABPD and ABLT pathways could represent a safeguard ensuring innervation to the EUS in case of upper nerve damage. Detailed knowledge of neuroanatomy and functional innervation of the urethra will enable more accurate animal models of neural development, disease, and dysfunction in the future.

Pudendal Nerve Stretch Reduces External Urethral Sphincter Activity in Rats

PURPOSE Most animal models of stress urinary incontinence simulate maternal injuries of childbirth since delivery is a major risk factor but they do not reproduce the nerve stretch known to occur during human childbirth. We hypothesized that pudendal nerve stretch produces reversible dysfunction of the external urethral sphincter. MATERIALS AND METHODS Female virgin Sprague-Dawley® rats were anesthetized with urethane. Bilateral pudendal nerve stretch or sham injury was performed for 5 minutes. External urethral sphincter electromyography and leak point pressure were recorded immediately before and after, and 10, 30, 60 and 120 minutes after pudendal nerve stretch. Post-pudendal nerve stretch results were compared to prestretch values and to values in sham injured animals. The pudendal nerves underwent qualitative histological assessment. The nucleus of Onuf was evaluated by immunohistochemistry and polymerase chain reaction for β-APP and c-Fos expression as markers of neuronal activity and injury. RESULTS A total of 14 rats underwent bilateral pudendal nerve stretch (9) or sham injury (5). Each nerve was stretched a mean ± SEM of 74% ± 18% on the left side and 63% ± 13% on the right side. Electromyography amplitude decreased significantly immediately after stretch compared to before stretch and after sham injury (p = 0.003) but it recovered by 30 minutes after stretch. There was no significant change in leak point pressure at any time. Two hours after injury histology showed occasional neuronal degeneration. β-APP and c-Fos expression was similar in the 2 groups. CONCLUSIONS Acute pudendal nerve stretch produces reversible electrophysiological dysfunction but without leak point pressure impairment. Pudendal nerve stretch shows promise in modeling injury. It should be tested as part of a multi-injury, chronic, physiological model of human childbirth injury.

Conversion of urodynamic pressures measured simultaneously by air-charged and water-filled catheter systems.

The objective of this study was to compare the simultaneous responses of water‐filled (WFC) and air‐charged (ACC) catheters during simulated urodynamic pressures and develop an algorithm to convert peak pressures measured using an ACC to those measured by a WFC.

Development Of An Area Equivalent Factor (aef) For Urethral Function

We developed a conductance formula for flow through rigid tubes that may be considered in the range for urethral opening important for connolling Fl. Tubes were tested by putting them at the bottom of a water column and letring the water run out of the coumn and recording Pr and Fl. Curve fitting of Pr and Fl data resulted in an exponential formula. A Propaionality constant was further determined to relate tubes of differing cross sectional area to an average curve fit formula. The final conductance formula, therefore, can be described as a cross sectional area equivalent factor (AEF) mm2 = 3.5 (FWPr0.58). that relates Fl and Pr to a rigid tube area. hliminary clinical observations suggest that the formula may have diagnostic potential for micturition dysfunction. Both the rate of urethral opening and maximal urethral opening may be useful information. Furthermore, a urethral compliance value, a ratio of opening area divided by the bladder Pr, may be useful in comparing patients with different Pr.

Direct bladder stimulation with percutaneous electrodes and impedance monitoring of volume in an SCI animal model.

Bladder responses to percutaneous electrodes were investigated with stimulation in three male spinal cats. The animals had been spinalized (T1 level lesion) 10 weeks prior to these studies and had been instrumented with chronic bladder had been spinalized (T1 level lesion) 10 weeks prior to these studies and had been instrumented with chronic bladder wall electrodes and suprapubic bladder catheters for filling and pressure recording. percutaneous stimulation in tethered animals was conducted wit hook electrodes inserted with a needle in the abdomen bilaterally adjacent to the bladder trigone. Stimulation was conducted with 40 Hz pulse trains of 10 to 30 mA for three seconds. Stimulation with both percutaneous and chronic electrodes induced high bladder pressures and voiding. In addition, with chronically implanted electrodes, impedance monitoring of bladder volume was found to be an effective recording technique.

Electrical stimulation of anal sphincter or pudendal nerve improves anal sphincter pressure.

OBJECTIVE: Stimulation of the pudendal nerve or the anal sphincter could provide therapeutic options for fecal incontinence with little involvement of other organs. The goal of this project was to assess the effects of pudendal nerve and anal sphincter stimulation on bladder and anal pressures. DESIGN: Ten virgin female Sprague Dawley rats were randomly allocated to control (n = 2), perianal stimulation (n = 4), and pudendal nerve stimulation (n = 4) groups. A monopolar electrode was hooked to the pudendal nerve or placed on the anal sphincter. Aballoon catheter was inserted into the anus to measure anal pressure, and a catheter was inserted into the bladder via the urethra to measure bladder pressure. Bladder and anal pressures were measured with different electrical stimulation parameters and different timing of electrical stimulation relative to spontaneous anal sphincter contractions. RESULTS: Increasing stimulation current had the most dramatic effect on both anal and bladder pressures. An immediate increase in anal pressure was observed when stimulating either the anal sphincter or the pudendal nerve at stimulation values of 1 mA or 2 mA. No increase in anal pressure was observed for lower current values. Bladder pressure increased at high current during anal sphincter stimulation, but not as much as during pudendal nerve stimulation. Increased bladder pressure during anal sphincter stimulation was due to contraction of the abdominal muscles. CONCLUSION: Electrical stimulation caused an increase in anal pressures with bladder involvement only at high current. These initial results suggest that electrical stimulation can increase anal sphincter pressure, enhancing continence control.