Interactive three-dimensional teaching models of the female and male pelvic floor.

Controversies regarding structure and function of the pelvic floor persist because of its poor accessibility and complex anatomical architecture. Most data are based on dissection. This "surgical" approach requires profound prior knowledge, because applying the scalpel precludes a "second look." The "sectional" approach does not entail these limitations, but requires segmentation of structures and three-dimensional reconstruction. This approach has produced several "Visible Human Projects." We dealt with limited spatial resolution and difficult-to-segment structures by proceeding from clear-cut to more fuzzy boundaries and comparing segmentation between investigators. We observed that the bicipital levator ani muscle consisted of pubovisceral and puborectal portions; that the pubovisceral muscle formed, together with rectococcygeal and rectoperineal muscles, a rectal diaphragm; that the external anal sphincter consisted of its subcutaneous portion and the puborectal muscle only; that the striated urethral sphincter had three parts, of which the middle (urethral compressor) was best developed in females and the circular lower ("membranous") best in males; that the rectourethral muscle, an anterior extension of the rectal longitudinal smooth muscle, developed a fibrous node in its center (perineal body); that the perineal body was much better developed in females than males, so that the rectourethral subdivision into posterior rectoperineal and anterior deep perineal muscles was more obvious in females; that the superficial transverse perineal muscle attached to the fibrous septa of the ischioanal fat; and that the uterosacral ligaments and mesorectal fascia colocalized. To facilitate comprehension of the modified topography we provide interactive 3D-PDFs that are freely available for teaching purposes. Clin. Anat. 33:275-285, 2020. © 2019 Wiley Periodicals, Inc.

Architecture of structures in the urogenital triangle of young adult males; comparison with females.

The fibro-muscular architecture of the urogenital triangle remains contentious. Reasons are small size of the constituting structures and poor visibility with most imaging methods. We reinvestigated the area in serial sections of three males (21-38 years old) of the American and Chinese Visible Human Projects and two 26-week-old male fetuses, and compared the findings with earlier observations in females. The mass of the levator ani muscle was approximately twofold smaller and its funnel shape steeper in males than females. In the levator hiatus, a strand of the smooth longitudinal muscle layer of the rectum, the 'rectourethral (RU) muscle', extended anteriorly from the anorectal bend to the penile bulb. Fibrous tissue that formed in the inferior reach of the fetal RU muscle identified the location of the developing perineal body (PB) and divided the muscle into posterior 'rectoperineal' and anterior 'deep perineal' portions. In males, the PB remained small and bipartite, so that the RU muscle presented as an undivided midline structure. The well-developed female PB, instead, intertwined with the deep perineal muscle and both structures passed the vagina bilaterally to form the perineal membrane in the posterior portion of the urogenital triangle. The urethral rhabdosphincter extended in the anterior portion of the urogenital triangle between the penile bulb inferiorly and the bladder neck superiorly, and consisted of a well-developed circular 'membranous' portion with bilateral posteroinferior 'wings' and a thinner 'prostatic' portion on the prostate anterior side. In men, muscles occupy the urogenital triangle, but additional tightening of the locally fibrous adipose tissue by the superficial transverse perineal muscle appears necessary to generate functional support in women. An interactive 3D pdf file with these anatomical details (available online) should allow more accurate interpretation of ultrasound, computed tomography and magnetic resonance images.

Architectural differences in the anterior and middle compartments of the pelvic floor of young-adult and postmenopausal females.

The pelvic floor guards the passage of the pelvic organs to the exterior. The near-epidemic prevalence of incontinence in women continues to generate interest in the functional anatomy of the pelvic floor. However, due to its complex architecture and poor accessibility, the classical 'dissectional' approach has been unable to come up with a satisfactory description, so that many aspects of its anatomy continue to raise debate. For this reason, we opted for a 'sectional' approach, using the Chinese Visible Human project (four females, 21-35 years) and the Visible Human Project (USA; one female, 59 years) datasets to investigate age-related changes in the architecture of the anterior and middle compartments of the pelvic floor. The puborectal component of the levator ani muscle defined the levator hiatus boundary. The urethral sphincter complex consisted of a circular proximal portion (urethral sphincter proper), a sling that passed on the vaginal wall laterally to attach to the puborectal muscle (urethral compressor), and a circular portion that surrounded the distal urethra and vagina (urethrovaginal sphincter). The exclusive attachment of the urethral sphincter to soft tissues implies dependence on pelvic-floor integrity for optimal function. The vagina was circular at the introitus and gradually flattened between bladder and rectum. Well-developed fibrous tissue connected the inferior vaginal wall with urethra, rectum and pelvic floor. With eight-muscle insertions, the perineal body was a strong, irregular fibrous node that guarded the levator hiatus. Only loose areolar tissue comprising a remarkably well developed venous plexus connecting the middle and superior parts of the vagina with the lateral pelvic wall. The posterolateral boundary of the putative cardinal and sacrouterine ligaments coincided with the adventitia surrounding the mesorectum. The major difference between the young-adult and postmenopausal pelvic floor was the expansion of fat in between the components of the pelvic floor. We hypothesize that accumulation of pelvic fat compromises pelvic-floor cohesion, because the pre-pubertal pelvis contains very little fibrous and adipose tissue, and fat is an excellent lubricant.

3D Topography of the Young Adult Anal Sphincter Complex Reconstructed from Undeformed Serial Anatomical Sections.

BACKGROUND: Pelvic-floor anatomy is usually studied by artifact-prone dissection or imaging, which requires prior anatomical knowledge. We used the serial-section approach to settle contentious issues and an interactive 3D-pdf to make the results widely accessible. METHOD: 3D reconstructions of undeformed thin serial anatomical sections of 4 females and 2 males (21-35y) of the Chinese Visible Human database. FINDINGS: Based on tendinous septa and muscle-fiber orientation as segmentation guides, the anal-sphincter complex (ASC) comprised the subcutaneous external anal sphincter (EAS) and the U-shaped puborectal muscle, a part of the levator ani muscle (LAM). The anococcygeal ligament fixed the EAS to the coccygeal bone. The puborectal-muscle loops, which define the levator hiatus, passed around the anorectal junction and inserted anteriorly on the perineal body and pubic bone. The LAM had a common anterior attachment to the pubic bone, but separated posteriorly into puborectal and "pubovisceral" muscles. This pubovisceral muscle was bilayered: its internal layer attached to the conjoint longitudinal muscle of the rectum and the rectococcygeal fascia, while its outer, patchy layer reinforced the inner layer. ASC contraction makes the ano-rectal bend more acute and lifts the pelvic floor. Extensions of the rectal longitudinal smooth muscle to the coccygeal bone (rectococcygeal muscle), perineal body (rectoperineal muscle), and endopelvic fascia (conjoint longitudinal and pubovisceral muscles) formed a "diaphragm" at the inferior boundary of the mesorectum that suspended the anorectal junction. Its contraction should straighten the anorectal bend. CONCLUSION: The serial-section approach settled contentious topographic issues of the pelvic floor. We propose that the ASC is involved in continence and the rectal diaphragm in defecation.

Anatomy and histology of the lower urinary tract.

The function of the lower urinary tract is basically storage of urine in the bladder and the at-will periodic evacuation of the stored urine. Urinary incontinence is one of the most common lower urinary tract disorders in adults, but especially in the elderly female. The urethra, its sphincters, and the pelvic floor are key structures in the achievement of continence, but their basic anatomy is little known and, to some extent, still incompletely understood. Because questions with respect to continence arise from human morbidity, but are often investigated in rodent animal models, we present findings in human and rodent anatomy and histology. Differences between males and females in the role that the pelvic floor plays in the maintenance of continence are described. Furthermore, we briefly describe the embryologic origin of ureters, bladder, and urethra, because the developmental origin of structures such as the vesicoureteral junction, the bladder trigone, and the penile urethra are often invoked to explain (clinical) observations. As the human pelvic floor has acquired features in evolution that are typical for a species with bipedal movement, we also compare the pelvic floor of humans with that of rodents to better understand the rodent (or any other quadruped, for that matter) as an experimental model species. The general conclusion is that the "Bauplan" is well conserved, even though its common features are sometimes difficult to discern.

Lack of specificity of commercially available antisera against muscarinergic and adrenergic receptors.

Commercially available antisera against five subtypes of muscarinic receptors and nine subtypes of adrenoceptors showed highly distinct immunohistochemical staining patterns in rat ureter and stomach. However, using the M(1-4) muscarinic receptor subtypes and alpha(2B)-, beta(2)-, and beta(3)-adrenoceptors as examples, Western blots with membranes prepared from cell lines stably expressing various subtypes of muscarinic receptors or adrenoceptors revealed that each of the antisera recognized a set of proteins that differed between the cell lines used but lacked specificity for the claimed target receptor. We propose that receptor antibodies need better validation before they can reliably be used.

The anatomical components of urinary continence.

BACKGROUND: The levator ani muscle (LAM) plays an important role in urinary continence, but the anatomical relationship between this pelvic floor muscle and the external urethral sphincter (EUS) remains incompletely understood. OBJECTIVE: To investigate the topographical relationship between the EUS and the LAM. DESIGN, SETTING, AND PARTICIPANTS: Serially sectioned and histochemically stained foetal pelves from eleven females and nine males (10-27 wk of gestation) were studied. Three foetal pelves (two female, 12 and 18 wk of gestation; one male, 12 wk of gestation) and three adult pelves (two females, 54 and 85 yr; one male, 75 yr) were stained immunohistochemically for the presence of striated and smooth muscle tissue. Three-dimensional reconstructions were prepared. MEASUREMENTS: Anatomy of the LAM and urethral sphincter components was evaluated qualitatively. RESULTS AND LIMITATIONS: The EUS has no direct bony attachment. In female foetuses, the inferior part of the EUS is firmly attached to the LAM by a tendinous connection. Contraction of this part of the EUS produces a force on the urethra in a posteroinferior direction. Contraction of the LAM compresses the rectum and moves the rectovaginal complex anteriorly and superiorly towards the urethra in a plane that lies parallel to, but superior of, that of the EUS. Simultaneous contraction of the LAM and EUS causes an anteriorly convex bend in the midurethra, which closes the midurethral lumen. A similar attachment of the EUS to the LAM is absent in the male. Our study is limited due to the absence of young adult study specimens. CONCLUSIONS: The EUS in females is anchored to the levator ani muscle via a tendinous connection. Because of this attachment to the LAM, proper function of the EUS is dependent on the integrity of the LAM and its attachment to the pelvic wall.

Lack of specificity of commercially available antisera: better specifications needed.

The ideal antiserum for immunohistochemical (IHC) applications contains monospecific high-affinity antibodies with little nonspecific adherence to sections. Many commercially available antibodies are "affinity" purified, but it is unknown if they meet "hard" specificity criteria, such as absence of staining in tissues genetically deficient for the antigen or a staining pattern that is identical to that of an antibody raised against a different epitope on the same protein. Reviewers, therefore, often require additional characterization. Although the affinity-purified antibodies used in our study on the distribution of muscarinic receptors produced selective staining patterns on sections, few passed the preabsorption test, and none produced bands of the anticipated size on Western blots. More importantly, none showed a difference in staining pattern on sections or Western blots between wild-type and knockout mice. Because these antibodies were used in most studies published thus far, our findings cast doubts on the validity of the extant body of morphological knowledge of the whole family of muscarinic receptors. We formulate requirements that antibody-specification data sheets should meet and propose that journals for which IHC is a core technique facilitate consumer rating of antibodies. "Certified" antibodies could avoid fruitless and costly validation assays and should become the standard of commercial suppliers.

Causes of fecal and urinary incontinence after total mesorectal excision for rectal cancer based on cadaveric surgery: a study from the Cooperative Clinical Investigators of the Dutch total mesorectal excision trial.

PURPOSE: Total mesorectal excision (TME) for rectal cancer may result in anorectal and urogenital dysfunction. We aimed to study possible nerve disruption during TME and its consequences for functional outcome. Because the levator ani muscle plays an important role in both urinary and fecal continence, an explanation could be peroperative damage of the nerve supply to the levator ani muscle. METHODS: TME was performed on cadaver pelves. Subsequently, the anatomy of the pelvic floor innervation and its relation to the pelvic autonomic innervation and the mesorectum were studied. Additionally, data from the Dutch TME trial were analyzed to relate anorectal and urinary dysfunction to possible nerve damage during TME procedure. RESULTS: Cadaver TME surgery demonstrated that, especially in low tumors, the pelvic floor innervation can be damaged. Furthermore, the origin of the levator ani nerve was located in close proximity of the origin of the pelvic splanchnic nerves. Analysis of the TME trial data showed that newly developed urinary and fecal incontinence was present in 33.7% and 38.8% of patients, respectively. Both types of incontinence were significantly associated with each other (P = .027). Low anastomosis was significantly associated with urinary incontinence (P = .049). One third of the patients with newly developed urinary and fecal incontinence also reported difficulty in bladder emptying, for which excessive perioperative blood loss was a significant risk factor. CONCLUSION: Perioperative damage to the pelvic floor innervation could contribute to fecal and urinary incontinence after TME, especially in case of a low anastomosis or damage to the pelvic splanchnic nerves.

The contribution of the levator ani nerve and the pudendal nerve to the innervation of the levator ani muscles; a study in human fetuses.

OBJECTIVES: The contributions of the pudendal and levator ani nerves to the innervation of the levator ani muscle (LAM) are disputed. Because of the relatively large size of the nerves in early life, we investigated this issue in human fetuses. METHODS: (Immuno)histochemically stained serial sections of nine human fetuses (9-22 wk of gestation) were investigated. Both the left and right sides of the fetal pelves were studied individually and 3D reconstructions were prepared. RESULTS: The levator ani nerve innervated the LAM in every pelvis, whereas a contribution of the pudendal nerve to the innervation of the LAM could be demonstrated in only 10 pelvic halves (56%). In 10 halves, we observed a communicating nerve branch between the pudendal and levator ani nerves that pierced the pelvic floor between the LAM and the coccygeus muscle. No sex differences were observed, but the innervation pattern did differ between the left and right side of a pelvis. CONCLUSIONS: The LAM often has a dual somatic innervation with the levator ani nerve as its constant and main neuronal supply.

Innervation of the pelvic floor muscles: a reappraisal for the levator ani nerve.

OBJECTIVE: We investigated the clinical anatomy of the levator ani nerve and its topographical relationship with the pudendal nerve. METHODS: Ten female pelves were dissected and a pudendal nerve blockade was simulated. The course of the levator ani nerve and pudendal nerve was described quantitatively. The anatomical data were verified using (immuno-)histochemically stained sections of human fetal pelves. RESULTS: The levator ani nerve approaches the pelvic-floor muscles on their visceral side. Near the ischial spine, the levator ani nerve and the pudendal nerve lie above and below the levator ani muscle, respectively, at a distance of approximately 6 mm from each other. The median distance between the levator ani nerve and the point of entry of the pudendal blockade needle into the levator ani muscle was only 5 mm. CONCLUSION: The levator ani nerve and the pudendal nerve are so close at the level of the ischial spine that a transvaginal "pudendal nerve blockade" would, in all probability, block both nerves simultaneously. The clinical anatomy of the levator ani nerve is such that it is prone to damage during complicated vaginal childbirth and surgical interventions.