The fiber type of the rabbits' striated external urethral sphincter: electrophoretic analysis of myosin

The fiber type of the rabbits' external urethral sphincter was investigated with O'Farrell's 2-dimensional electrophoresis of constituent proteins of glycerinated muscle, peptide mapping of myosin heavy chains and non-denaturing pyrophosphate gel electrophoresis of myosin. It was determined as fast type by all three methods. The relative proportions of muscle fibers which were estimated from myosin light chain contents were 88 per cent fast type and 12 per cent slow type. On the basis of the results presented, it was shown to be fast twitch muscle, but not identical to the psoas muscle.     

The proportions of fiber types in human external urethral sphincter: electrophoretic analysis of myosin

Summary The proportions of fast and slow myosin molecules in external urethral sphincter specimens from ten urodynamically normal male bladder carcinoma patients were estimated from the contents of fast and slow myosin light chains in two-dimensional electrophoretic gels. The percentages of fast and slow myosin molecules ranged from 5.0% to 61.4% with a mean of 35.5% and from 38.6% to 95.0% with a mean of 65.5% respectively. It is therefore concluded that the human external urethral sphincter is composed of both fast and slow muscle fibers as well as other voluntary muscles. The human external urethral sphincter is considered to be a highly fatigue-resistant muscle with a very high proportion of slow muscle fibers. In the cases studied so far, there is a great diversity in the proportions of fast and slow myosin molecules; the reason for this remains unknown.     

Biochemical analysis of the external urethral sphincter of female rabbits]

The proportion of fast and slow myosin molecules in external urethral sphincter (EUS) was studied. The contents of fast and slow myosin light chains in EUS from 6 female rabbits (Japanese White, 6-month-old) were estimated using two-dimensional electrophoresis. The percentages of fast and slow myosin molecules were 54.5 +/- 14.9 and 46.0 +/- 14.9, respectively. It was thus shown that female rabbits EUS is composed of both fast and slow muscle fibers. The ratio of fast to slow myosins in female EUS was different from that in male EUS (fast myosin 90%). These results implicate that female and male EUS would be functionally distinct.     

Electrophoretic and ultrastructural analysis of the rabbit's striated external urethral sphincter

The male rabbit's external urethral sphincter was investigated with O'Farrell's 2-dimensional electrophoretic analysis of myosin light chains and electron microscopy. Its pattern of myosin light chains was different from that of the soleus (predominantly slow twitch muscle) but was very similar to that of the psoas (predominantly fast twitch muscle). Ultrastructurally it was shown to be red muscle resembling the soleus. Therefore the fiber type of the rabbit's external urethral sphincter was determined to be the red (fast) type.     

Proportions of fiber types in the external urethral sphincter of young nulliparous and old multiparous rabbits

Summary The proportions of fast and slow myosin molecules in specimen's of the external urethral sphincter (EUS) from 6 young nulliparous (6-month-old) and 6 old multiparous (2-year-old) rabbits were studied using myosin heavy chain electrophoresis. The percentages of fast and slow myosin molecules were 80.4±4.5% and 19.6±4.5% in the EUS from nulliparas, and 68.7±6.3% and 31.3±6.3% in the EUS from multiparas. The difference between the two groups was significant (P<0.01). We suggest that a selective decrease in the volume of type 2 (fast) muscle fibers and/or conversion of type 2 to type 1 (slow) muscle fibers had taken place in the EUS of old multiparas. The proportional change in the constitutent muscle fibers of the EUS with aging may play a role in human genuine stress incontinence.     

A comparative study of the human external sphincter and periurethral levator ani muscles

Specimens from the human male and female external urethral sphincter and the periurethral levator ani muscle have been examined using histochemical and electron microscopic techniques. In both sexes the external sphincter consists of a single population of type I (slow twitch) fibres with a mean diameter of 17.47 +/- 0.7 micrometers in the absence of muscle spindles. In contrast, the periurethral levator ani possesses muscle spindles and the constituent fibres form a heterogeneous population of type I and type II (fast twitch) fibres, with mean diameters of 45.5 +/- 0.8 micrometer and 59.5 +/- 3.4 micrometers respectively. These findings indicate that the external urethral sphincter is functionally adapted to maintain tone over prolonged periods and may be of considerable importance in producing active urethral closure during continence. The anatomical location and fibre characteristics of the levator ani muscle suggest that these fibres actively assist in urethral closure, particularly during events which cause elevation of intra-abdominal pressure. In view of the differences in fibre characteristics between the external urethral sphincter and the levator ani, EMG activity recorded from a single site in the levator ani may not be representative of the functional status either of other levator ani muscle fibres or of the external urethral sphincter.     

Histochemical study of external urethral sphincter in the rabbit. Analysis with construction of histograms

Male rabbit's external urethral sphincter was examined by histochemical muscle fiber typing (myosin ATP-ase staining), and the analysis with construction of histograms regarding to muscle fiber types were performed. Rabbit's external urethral sphincter was predominantly composed of fast twitch (type 2) fibers (87.3%) as a whole. But the proportion of constituent fiber types varied according to the layers, i.e., the slow twitch (type 1) fibers constituted a relatively high percentage (33.4%) in the inner third layer, while few of the type 1 fibers were found in the outer third layer. The all histograms regarding to fiber type in different layers were normal bell-shaped distribution curves. The mena diameter of type 2 fibers (14.7 microns) was evidently larger than that of type 1 fibers (20.5 microns). All three kinds of muscle fibers equally tended to increase in size toward the outer direction, and in every three layers, the diameter of type 2 was larger than that of type 1 also. The definite differences in the proportion of fiber types and fiber sizes between layers may implicate that the inner and outer layers play different roles, i.e., continuous tonic constriction in the former and sporadic strong constriction of short duration in the latter, under different neural regulations. As far as rabbit's external urethral sphincter is concerned, sporadic strong constriction should be mainly dependent on the muscle fibers of large size composing the outer layer, especially the fast twitch fibers. It is possible that the rabbit is so adapted that it could interrupt urination promptly.     

Can simultaneous perineal sonography and urethrocystometry help explain urethral pressure variations?

Objective: To correlate urodynamic with perineal sonographic findings in pressure variations. Patients and methods: In 15 women presenting with urethral pressure variations a urodynamic evaluation with water filling cystometry, urethral pressure at rest and during coughing and uroflowmetry were performed. During water filling cystometry, there were simultaneous perineal video-sonography and urethrocystometry. Video ultrasound images and urodynamic curves were simultaneously monitored on a computer screen. Results: Simultaneous ultrasound and urodynamic evaluation in the 15 patients revealed movements in two areas leading to urethral pressure variations: activity of the pelvic floor muscles and of the urethral sphincter muscles. For the pelvic floor, we found either slow or fast contractions with, respectively, slow (15–30 cm H₂O for 3–10 sec) or fast (30–130 cm H₂O for 1–3 sec) urethral pressure changes. Urethral sphincter contractions were always fast, resulting in fast pressure changes of 30–170 cm H₂O for 1–3 sec. Conclusion: Evaluation of simultaneous perineal sonography and urethrocystometry shows the association of urethral pressure variations and muscle activity. Urethral pressure variations are caused by the activity of urethral sphincter or pelvic floor muscles. With ultrasound the activity of the urethral sphincter muscle can directly be seen whereas pelvic floor muscle activity is indirectly visible. Pelvic floor muscle contractions are either fast or slow, whereas the urethral sphincter muscle contractions are always fast contractions.     

Histochemical study of human external urethral sphincter]

Human external urethral sphincters (n = 13) were studied histochemically, using ATP-ase staining. The proportions of constituent muscle fiber types (slow twitch type 1 fibers: 35.6-97.7%, mean 65.7%, SD 16.6) were different among the individuals. There were no significant relationships between the percentage of type 1 fibers and the sex or the age. As for the mean muscle fiber diameter of 13 cases, the sizes of type 2 fibers (19.2-42.4 um, mean 27.8 um) were significantly (p less than 0.05, t-test) larger than those of type 1 fibers (15.7-30.3 um, mean 22.3 um). In analyses of individual cases, 10 male cases had significantly larger type 2 fibers than type 1. Two of 3 female cases had no significant difference in diameter between the two fiber types and the remaining 1 female case, in contrast with male cases, had significantly larger type 1 fibers than type 2. There was no significant relationship between the proportions and the diameters of constituent muscle fiber types. Our study thus showed extreme interindividual variation and implicated the presence of sexual difference in human external urethral sphincter muscle.     

An anatomical description of the male and female urethral sphincter complex

PURPOSE: We performed a detailed study of the lower urinary tract of the male and female human fetus to elucidate the anatomy of the urethral sphincter complex in both sexes and its relationship to the surrounding organs and tissues. MATERIALS AND METHODS: A total of 12 male and 14 female normal human pelvic specimens ranging from 17.5 to 38 weeks of gestation were studied by serial sections and immunohistochemical analysis. Three-dimensional reconstructions were created from serial sections to demonstrate the anatomy of the lower urogenital tract and urethral sphincter in both sexes. Specific attention was directed to the sphincteric muscle of the urethra. RESULTS: The urinary continence mechanism is formed by a combination of detrusor, trigone and urethral sphincter muscles with distinctive histological characteristics in both sexes. In males the external urethral sphincter covers the ventral surface of the prostate as a crescent shape above the verumontanum, horseshoe shape below the verumontanum and crescent shape along the proximal bulbar urethra. The levator ani muscles form an open circle around the external sphincter with a hiatus at the ventral aspect. In females the external urethral sphincter covers the ventral surface of the urethra in a horseshoe shape. Caudally the same horseshoe-shaped external sphincter increases in size to envelop the distal vagina. The levator ani muscles do not support the proximal urethra. The smooth and striated muscle components of the urethral sphincter complex are inseparable in both sexes. CONCLUSIONS: The developmental anatomy of the urethral sphincter complex is analogous in both sexes. The male and female urinary sphincter mechanism is composed of detrusor, trigone and urethral muscles, each of different muscular origins. The levator ani does not surround the ventral aspect of the urethra and may not have an active role in continence in both sexes. This new concept in the anatomy of male and female sphincter morphology may help to refine our reconstructive and ablative surgical techniques.     

THE ‘UROGENITAL DIAPHRAGM’, EXTERNAL URETHRAL SPHINCTER AND RADICAL PROSTATECTOMY

Background : The present study was performed to determine whether a ‘urogenital diaphram’ exists, to examine the true nature of the striated external urethral sphincter and to evaluate whether the standard technique for radical prostatectomy damages the external sphincter. Methods : Fifty radical prostatectomies were performed using optical magnification and the dorsal bunching technique, and the external sphincter was carefully examined. Ten human cadavers and one 5-year-old baboon were dissected with longitudinal (sagittal) and transverse sections being taken through the prostate apex, membranous and bulbar urethrae. During the standard technique for dorsal vein control during radical prostatectomy, the tissue incorporated within the ligature was examined for striated muscle. Results : No ‘urogenital diaphram’ could be demonstrated in any human or baboon tissue. The striated external urethral sphincter is a cylinder of muscle surrounding the membranous urethra, extending from the perineal membrane to the prostate and continuing over the prostate as part of the anterior fibromuscular stroma. Striated muscle was present in the ligated material from the dorsal venous complex. Conclusions : The ‘urogenital diaphram’ is a myth. The standard technique of radical prostatectomy significantly damages the external sphincter.     

Further characterization of the external urethral sphincter in spinal cord injury: study during spinal shock and evolution of responsiveness to alpha-adrenergic stimulation

To shed further insight into its ever evolving concepts, we studied the activity of the external urethral sphincter in patients with spinal cord injury. Study during the phase of acute spinal shock revealed persistent electromyographic activity in the external urethral sphincter and no activity in the external anal sphincter, suggesting the presence of functional dissociation between the 2 sphincters from the onset of acute spinal injury. The genesis of dissociated activity was discussed by reference to recent experiments on the individual differences in motor neuron and muscle subtypes. Only the external urethral sphincter of chronic paraplegics demonstrated a uniformly increased electromyographic activity after alpha-adrenergic stimulation, while the response in the external anal sphincter was variable. The mechanism of increased response to alpha-adrenergic stimulation in the denervated external urethral sphincter appears to reside either in direct postsynaptic stimulation or recruitment of motor neurons other than the somatic pudendal system, since pre-treatment with competitive muscle relaxant failed to block this increased response in the external urethral sphincter.     

The effect of external urethral sphincter contraction on the cavernosus muscles and its role in the sexual act

Objectives/Aim A study of the effect of external urethral sphincter contraction on ischio-/bulbo-cavernosus muscles could not be traced in the literature. We investigated the hypothesis that external urethral sphincter contraction induces cavernosus muscles’ contraction. Methods Twenty-one healthy volunteers (age 37.6 ± 9.7 SD years, 12 men, nine women) were studied. The electromyographic response of the ischio- and bulbo-cavernosus muscles to external urethral sphincter stimulation was recorded before and after anesthetization of the external urethral sphincter, and the ischio- and bulbo-cavernosus muscles; the response was also recorded using normal saline instead of lidocaine. Results Upon external urethral sphincter stimulation (five square pulses, 1 ms duration, 53.8 ± 10.2 mA threshold), both cavernosus muscles exhibited increased electromyographic activity with a mean amplitude of 386.2 ± 44.9 μV for the ischio-cavernosus and 318.4 ± 36.6 μV for the bulbo-cavernosus muscle. The mean latency read 16.8 ± 1.3 ms for the ischio-cavernosus muscle and 15.7 ± 1.2 ms for the bulbo-cavernosus muscle. Neither the ischio- nor the bulbo-cavernosus muscle responded to stimulation of the anesthetized external urethral sphincter, but both responded after saline administration. Likewise, the anesthetized ischio- and bulbo-cavernosus muscles showed no response to external urethral sphincter stimulation but responded after saline had been injected. Conclusions Increased electromyographic activity of the two cavernosus muscles on external urethral sphincter stimulation presumably denotes contraction of these two muscles and that this action is probably reflex, mediated through the ‘sphinctero-cavernosus-reflex’. Cavernosus muscles’ contraction assists in the erectile and ejaculatory mechanisms. It is suggested that this reflex be included as an investigative tool in the diagnosis of erectile and ejaculatory disorders, provided further studies are performed.     

A study of the continence mechanism of the external urethral sphincter with identification of the voluntary urinary inhibition reflex

PURPOSE: The role of the external urethral sphincter in the opposition and interruption of the act of voiding was investigated. MATERIALS AND METHODS: The study included 7 men and 5 women with a mean age plus or minus standard deviation of 38.6 +/- 11.2 years. The bladder was filled with saline up to the urge sensation. Detrusor and posterior urethral pressures were recorded before and upon resisting the reflex detrusor contraction, and upon interrupting voiding by voluntary external urethral sphincter contraction. The test was repeated by interrupting the urinary stream with external urethral sphincter electrostimulation. The electromyography response of the internal urethral sphincter to the suppression and interruption of voiding was documented before and after internal urethral sphincter anesthetization. RESULTS: Suppression of the reflex detrusor contraction as well as of urinary stream interruption by external urethral sphincter contraction voluntarily or by electrostimulation resulted in a significant detrusor pressure decrease (p <0.01) and urethral pressure increase (p <0.001). Internal urethral sphincter electromyography activity, which normally disappears during voiding, was still present. After internalurethral sphincter anesthetization subjects suppressed the reflex detrusor contraction by voluntary external urethral sphincter contraction for a mean of 62.6 +/- 9.6 seconds, after which involuntary voiding occurred. The internal urethral sphincter showed no electromyography activity. CONCLUSIONS: The external urethral sphincter induces continence by preventing internal urethral sphincter relaxation at the detrusor contraction, which is suggested to be reflex in nature and is called the voluntary urinary inhibition reflex, and by mechanically compressing the urethra. Contraction of the external urethral sphincter, which is a striated muscle, mechanically occludes the urethra for a few seconds, by which time the detrusor has relaxed as an effect of the voluntary inhibition reflex.     

The motor nerve supply of the external urethral sphincter muscles: An electrophysiologic study

The term “intrinsic external urethral sphincter” has recently been applied to the striated muscle immediately surrounding the membranous urethra, thus distinguishing and separating it from the periurethral striated muscle which is a component of the pelvic floor. The innervation of the intrinsic external urethral sphincter is still controversial. Six male patients with a sustained spinal cord lesion above D-4 underwent electrophysiological evaluation of the reflex and direct evoked responses to stimulation of the pudendal nerve branches in the perineal region. Recording of the motor unit potentials was performed using a catheter-mounted concentric needle introduced into the intrinsic external urethral sphincter transurethrally. The results of this study indicate that the pudendal nerve, i.e., somatic, plays an important role in the innervation of the intrinsic external urethral muscle. It does not, however, exclude the possibility that the autonomic nervous system also innervates this muscle.     

Adrenergic innervation of the striated muscle of the intrinsic external urethral sphincter from patients with lower motor spinal cord lesion

The adrenergic innervation of smooth and striated muscle components of the intrinsic external urethral sphincter from patients with suprasacral lesions and detrusor-sphincter dyssynergia has been described previously, when no adrenergic nerves were found associated with striated muscle fibers. In our study the intrinsic external urethral sphincter from patients with lower motor neuron lesions and detrusor areflexia was studied histochemically using the glyoxylic acid method to visualize catecholamines. Varicose adrenergic nerves were demonstrated in the smooth muscle. Adrenergic nerve fibers also were found along the edge of individual striated muscle fibers as well as around striated muscle bundles. Blood vessels in both regions of the urethral sphincter were innervated by adrenergic nerves. We conclude that in patients with lower motor neuron lesions and detrusor areflexia there is a substantial invasion by adrenergic nerve fibers in relation to smooth and striated muscle in the urethra, although the function of the nerve fibers is not known.     

Innervation of the female human urethral sphincter: 3D reconstruction of immunohistochemical studies in the fetus

OBJECTIVES: The precise location, origin and nature of nerve fibers innervating the urethral sphincter have not been clearly established. Classical anatomical studies based on cadaver dissections have provided conflicting results concerning the location of pudendal and autonomic nerve fibers. This study was designed to identify nerve fibers innervating the urethral sphincter and to provide a three-dimensional representation of their tissue relations in the female human fetus. MATERIALS AND METHODS: Histology and immunohistochemistry (Masson's Trichromic, Luxol Fast Blue, Protein S 100 immunostaining and smooth fiber actin immunostaining) were performed on the external urethral sphincter of ten female fetuses with a crown-rump length of 112 to 340mm. Three-dimensional reconstructions of the urethral structure and innervation were obtained from serial sections using Surf Driver 3.5.3 software (David Moody and Scott Lozanoff). RESULTS: Three-dimensional reconstructions of the same sections with different stains demonstrated the precise structure of the muscle layers (smooth and striated muscle fibers) and nerve fibers (myelinated and unmyelinated) and their relations with the urethra and vaginal wall. The proximal third consisted of a circular smooth muscle sphincter, the middle third consisted of two circular layers of smooth and striated muscle fibers and the distal third consisted of a circular layer of smooth muscle fibers surrounded by an omega-shaped layer of striated muscle fibers. In the proximal third of the urethral sphincter, myelinated fibers were identified running with unmyelinated fibers from the pelvic plexus. These fibers were closely related to the lateral and anterior aspects of the vagina. Unmyelinated fibers entered the smooth muscle part of the sphincter at 4 o'clock and at 8 o'clock. Most myelinated fibers entered the sphincter at 3 o'clock and at 9 o'clock. CONCLUSION: Histological and immunohistochemical three-dimensional reconstruction of the anatomical structures of the urethral sphincter provides a better understanding of the origin and nature of the Innervation participating in urinary continence. It provides a very informative view of the three-dimensional arrangement of sphincter muscle layers.     

The development of the external urethral sphincter in humans

OBJECTIVE: To assess the hypothesis that during fetal development, the external urethral sphincter changes from a concentric sphincter of undifferentiated muscle fibres to a transient ring of striated muscle which regresses caudo-cranially in the posterior urethra during the first year of life, when the sphincter assumes its omega-shaped configuration. MATERIALS AND METHODS: The anatomy and development of the external urinary sphincter was assessed in human males and females during fetal life. Plastic-embedded sections (transverse, sagittal and frontal planes; 300-700 microm) of the pelvis of 31 females and 31 males (9 weeks of gestation to newborn) were stained with azure II/methylene blue/basic fuchsin and viewed at x 4-80. The sections of interest were taken from the bladder neck to the perineum. The sections of the membranous urethra were reconstructed three-dimensionally using a computer program. RESULTS: In both male and female an omega-shaped external sphincter was apparent in all specimens at > 10 weeks of gestation. In the early fetal period (ninth week), there was undifferentiated mesenchyme; in this period the mesenchyme was more dense in the anterior part and loose in the posterior part of the urethra. In females, there was a close connection between the urethra and the anterior wall of the vagina. CONCLUSION: The omega-shaped configuration of the external urethral sphincter was recognisable from 10 weeks of gestation in both sexes. There was no suggestion of a change from a cylindrical to an omega-shaped sphincter in the fetal period to birth. Also, a transient 'tail' posterior to the sphincter was not apparent. The rectovesical septum was well developed in neonates. There is no reason to assume that the development of the septum leads to an apoptosis of muscle cells in the posterior part of the external sphincter in males after birth. The anatomical development of the external sphincter does not explain transient outlet obstruction during fetal life. The function of the muscle may change during development because of neuronal maturation.     

Urethral function in neurological disorders of the lower urinary tract

Simultaneous measurements of intravesical pressure, urethral pressure, and electromyographic activity of perianal muscles were performed during bladder filling and voiding in 56 patients with neurogenic lower urinary tract dysfunction. In 21 patients simultaneous measurements were taken of the EMG activity of the striated-muscle external urethral sphincter. During bladder filling the urethral pressure was found to correspond to the EMG activity of both muscles only in those patients with normal urethral pressure. During voiding the EMG activity of neither muscle corresponded well with changes in urethral pressure. The EMG activity of perianal muscles corresponded to the activity of the external urethral sphincter during filling of the bladder but not during voiding.     

Interference pattern in perineal muscles: A quantitative electromyographic study in patients before and after transurethral surgery of the prostate

The study aimed at assessing alterations in muscular activity in the external urethral sphincter when the internal sphincter located at the bladder neck was resected during TUR-P, and at determining whether activity in the external urethral sphincter increased to compensate for the loss of the internal sphincter. Perineal muscles were examined with quantitative EMG recordings, including interference pattern and fiber density before and after surgery. Fiber density increased in the external urethral sphincter after surgery. This indicates a reinnervation in the muscle, probably due to a peripheral nerve lesion that occurs during TUR-P, and may also explain the reduction in penile erectibility observed after surgery. The lack of compensatory activity in the external urethral sphincter expressed as unchanged number of turns may be explained as a disturbed feedback mechanism and a decreased central activation or to the lithotomy position at examination. The internal part of the external sphincter not available for measurement may compensate for the loss in bladder-neck sphincter function. Neurourol. Urodynam. 16:101–109, 1997. © 1997 Wiley-Liss, Inc.