Development of the human prepuce and its innervation.

Development of the human prepuce was studied over the course of 9-17 weeks of gestation in 30 specimens. Scanning electron microscopy revealed subtle surface features that were associated with preputial development, namely the appearance of epidermal aggregates that appeared to be associated with formation of the preputial fold. Transverse and sagittal sections revealed that the epidermis of the glans is considerably thicker than that of the penile shaft. We described a novel morphogenetic mechanism of formation of the preputial lamina, namely the splitting of the thick epidermis of the glans into the preputial lamina and the epidermis via the intrusion of mesenchyme containing red blood cells and CD31-positive blood vessels. This process begins at 10-11 weeks of gestation in the proximal aspect of the glans and extends distally. The process is likely to be androgen-dependent and mediated via androgen receptors strategically localized to the morphogenetic process, but signaling through estrogen receptor may play a role. Estrogen receptor alpha (ESR1) has a very limited expression in the developing human glans and prepuce, while estrogen receptor beta (ESR2) is expressed more broadly in the developing preputial lamina, epidermis and urethra. Examination of the ontogeny of innervation of the glans penis and prepuce reveals the presence of the dorsal nerve of the penis as early as 9 weeks of gestation. Nerve fibers enter the glans penis proximally and extend distally over several weeks to eventually reach the distal aspect of the glans and prepuce by 14-16 weeks of gestation.

Development of the human penis and clitoris.

The human penis and clitoris develop from the ambisexual genital tubercle. To compare and contrast the development of human penis and clitoris, we used macroscopic photography, optical projection tomography, light sheet microscopy, scanning electron microscopy, histology and immunohistochemistry. The human genital tubercle differentiates into a penis under the influence of androgens forming a tubular urethra that develops by canalization of the urethral plate to form a wide diamond-shaped urethral groove (opening zipper) whose edges (urethral folds) fuse in the midline (closing zipper). In contrast, in females, without the influence of androgens, the vestibular plate (homologue of the urethral plate) undergoes canalization to form a wide vestibular groove whose edges (vestibular folds) remain unfused, ultimately forming the labia minora defining the vaginal vestibule. The neurovascular anatomy is similar in both the developing human penis and clitoris and is the key to successful surgical reconstructions.

Human glans and preputial development.

The urethra within the human penile shaft develops via (1) an "Opening Zipper" that facilitates distal canalization of the solid urethral plate to form a wide urethral groove and (2) a "Closing Zipper" that facilitates fusion of the epithelial surfaces of the urethral folds. Herein, we extend our knowledge by describing formation of the human urethra within the glans penis as well as development of the prepuce. Forty-eight normal human fetal penile specimens were examined using scanning electron microscopy and optical projection tomography. Serial histologic sections were evaluated for morphology and immunohistochemical localization for epithelial differentiation markers: Cytokeratins 6, 7, 10, FoxA1, uroplakin and the androgen receptor. As the closing zipper completes fusion of the urethral folds within the penile shaft to form a tubular urethra (~ 13 weeks), canalization of the urethral plate continues in proximal to distal fashion into the glans penis to directly form the urethra within the glans without forming an open urethral groove. Initially, the urethral plate is attached ventrally to the epidermis via an epithelial seam, which is remodeled and eliminated, thus establishing mesenchymal confluence ventral to the glanular urethra. The morphogenetic remodeling involves the strategic expression of cytokeratin 7, FoxA1 and uroplakin in endodermal epithelial cells as the tubular glanular urethra forms. The most ventral epithelial cells of the urethral plate are pinched off from the glanular urethra and are reabsorbed into the epidermis ultimately losing expression of their markers, a process undoubtedly regulated by androgens. The prepuce initially forms on the dorsal aspect of the glans at approximately 12 weeks of gestation. After sequential proximal to distal remodeling of the ventral urethral plate along the ventral aspect of glans, the prepuce of epidermal origin fuses in the ventral midline.

Epithelial-mesenchymal transformation and apoptosis in rat urethra development.

BackgroundTo examine the mechanism of urethral seam formation during embryonal development of rat urethra.MethodsTime-mated Sprague-Dawley rats were killed and the genital tubercles of male pups harvested on embryonic day (ED) 15, 16, 18, and 19. External morphology was observed under scanning electron microscope. Serial transverse sections were prepared to examine dynamic changes in the urethral seam morphology with hematoxylin-eosin staining, immunohistochemistry, transmission electron microscopy, and double immunofluorescence.ResultsBilateral outgrowth of urethral swelling followed by urethral plate fusion in the midline to form urethral seam was observed from ED 16 onwards. Coexpression of epithelial and mesenchymal markers was observed in several cells at the urethral seam; a few cells with coexpression of epithelial and apoptotic markers were also observed. Mesenchymal transformation of epithelial cells and apoptotic epithelial cells was observed under transmission electron microscope.ConclusionUrethral formation occurs by tubulogenesis, which initiates proximally and progresses distally. This is the first study to demonstrate epithelial-mesenchymal transformation and epithelial cell apoptosis in the urethral seam cells of fetal rats. These findings provide new insights into the mechanisms involved in embryonal development of the urethra.

Structural Reorganization of the Vaginal Mucosa in Stress Urinary Incontinence under Conditions of Er:YAG Laser Treatment.

Structural characteristics of the vaginal mucosa in stress incontinence and its correction by IncontiLase technology were studied. Studies of vaginal biopsy specimens before the exposure showed degenerative and atrophic changes in the stratified squamous epithelium, disorganization of fibrillar structures of the intercellular matrix, and microcirculatory disorders. Studies after Er:YAG laser exposure showed signs of neocollagenogenesis and elastogenesis, foci of neoangiogenesis, reduction of epithelial degeneration and atrophy, and an increase of the fibroblast population. Morphometry showed that the volume density of blood capillaries and the thickness of the epithelial layer increased by 61.1 and 64.5%, respectively. The use of IncontiLase technology in stress incontinence led to structural reorganization of the vaginal mucosa, improving its morphology and function and alleviating the symptoms of incontinence.

Flutamide-induced hypospadias in rats: A critical assessment.

This paper provides the first detailed description of flutamide-induced hypospadias in the rat based upon wholemount, histologic, three-dimensional reconstruction, scanning electron microscopic, and immunocytochemical analysis. The penile malformations elicited by this potent anti-androgen include a substantial proximal shift in the urethral meatus that clearly conforms to the definition of hypospadias based upon specific morphological criteria for this malformation. Through examination of the normal penile development and flutamide-induced abnormal penile development observed in prenatally oil- and flutamide-treated rats, our analysis provides insights into the morphogenetic mechanism of development of hypospadias. In this regard, a common theme in normal penile development is midline fusion of epithelia followed by removal of the epithelial seam and establishment of midline mesenchymal confluence during development of the penile urethra and prepuce, processes which are impaired as a result of prenatal flutamide treatment. The developmental processes occurring in normal penile development, through comparison with development of female external genitalia and those impaired due to prenatal flutamide treatment, are consistent with critical role of androgen receptors in normal penile development in the rat, and the specific penile abnormalities embodied in flutamide-induced rat hypospadias.

Topographic distribution of serotonin-immunoreactive urethral endocrine cells and their relationship with calcitonin gene-related peptide-immunoreactive nerves in male rats.

We investigated the topographic distribution and morphology of serotonin (5-HT)-immunoreactive endocrine cells in the urethra of male rats, and focused on their relationship with peptidergic nerve fibers immunoreactive for calcitonin gene-related peptide (CGRP). Urethral endocrine cells immunoreactive for 5-HT were densely distributed in the epithelial layers of the prostatic part, but were sparsely distributed in the membranous and spongy parts of urethra. Distribution of urethral endocrine cells with 5-HT immunoreactivity in the prostatic part was restricted from the internal urethral orifice to the region of seminal colliculus. 5-HT-immunoreactive endocrine cells were also observed in the ductal epithelial layers of coagulating glands, prostatic glands, and seminal vesicles. 5-HT-immunoreactive endocrine cells were triangular or flask in shape and possessed an apical projection extending toward the urethral lumen, and basal or lateral protrusions intruding between other epithelial cells were also detected in some cells. Double immunolabeling for 5-HT and CGRP revealed that CGRP-immunoreactive nerve fibers attached to urethral endocrine cells with 5-HT immunoreactivity in the prostatic part. These results suggest that urethral endocrine cells may release 5-HT in response to luminal stimuli, and that these cells and CGRP-immunoreactive nerves may regulate each other by an axon reflex mechanism.

Purified Human Skeletal Muscle-Derived Stem Cells Enhance the Repair and Regeneration in the Damaged Urethra.

BACKGROUND: Postoperative damage of the urethral rhabdosphincter and nerve-vascular networks is a major complication of radical prostatectomy and generally causes incontinence and/or erectile dysfunction. The human skeletal muscle-derived stem cells, which have a synchronized reconstitution capacity of muscle-nerve-blood vessel units, were applied to this damage. METHODS: Cells were enzymatically extracted from the human skeletal muscle, sorted using flow cytometry as CD34/45 (Sk-34) and CD29/34/45 (Sk-DN/29) fractions, and separately cultured/expanded in appropriate conditions within 2 weeks. Urethral damage was induced by manually removing one third of the wall of the muscle layer in nude rats. A mixture of expanded Sk-34 and Sk-DN/29 cells was applied on the damaged portion for the cell transplantation (CT) group. The same amount of media was used for the non-CT (NT) group. Urethral pressure profile was evaluated via electrical stimulation to assess functional recovery. Cell engraftments and differentiations were detected using immunohistochemistry and immunoelectron microscopy. Expression of angiogenic cytokines was also analyzed using reverse transcriptase-polymerase chain reaction and protein array. RESULTS: At 6 weeks after transplantation, the CT group showed a significantly higher functional recovery than the NT group (70.2% and 39.1%, respectively; P < 0.05). Histological analysis revealed that the transplanted human cells differentiated into skeletal muscle fibers, nerve-related Schwann cells, perineuriums, and vascular pericytes. Active paracrine angiogenic cytokines in the mixed cells were also detected with enhanced vascular formation in vivo. CONCLUSIONS: The transplantation of Sk-34 and Sk-DN/29 cells is potentially useful for the reconstitution of postoperative damage of the urethral rhabdosphincter and nerve-vascular networks.

Complex epithelial remodeling underlie the fusion event in early fetal development of the human penile urethra.

We recently described a two-step process of urethral plate canalization and urethral fold fusion to form the human penile urethra. Canalization ("opening zipper") opens the solid urethral plate into a groove, and fusion ("closing zipper") closes the urethral groove to form the penile urethra. We hypothesize that failure of canalization and/or fusion during human urethral formation can lead to hypospadias. Herein, we use scanning electron microscopy (SEM) and analysis of transverse serial sections to better characterize development of the human fetal penile urethra as contrasted to the development of the human fetal clitoris. Eighteen 7-13 week human fetal external genitalia specimens were analyzed by SEM, and fifteen additional human fetal specimens were sectioned for histologic analysis. SEM images demonstrate canalization of the urethral/vestibular plate in the developing male and female external genitalia, respectively, followed by proximal to distal fusion of the urethral folds in males only. The fusion process during penile development occurs sequentially in multiple layers and through the interlacing of epidermal "cords". Complex epithelial organization is also noted at the site of active canalization. The demarcation between the epidermis of the shaft and the glans becomes distinct during development, and the epithelial tag at the distal tip of the penile and clitoral glans regresses as development progresses. In summary, SEM analysis of human fetal specimens supports the two-zipper hypothesis of formation of the penile urethra. The opening zipper progresses from proximal to distal along the shaft of the penis and clitoris into the glans in identical fashion in both sexes. The closing zipper mechanism is active only in males and is not a single process but rather a series of layered fusion events, uniquely different from the simple fusion of two epithelial surfaces as occurs in formation of the palate and neural tube.

Rat animal model for preclinical testing of microparticle urethral bulking agents.

OBJECTIVES: To develop an economic, practical and readily available animal model for preclinical testing of urethral bulking therapies, as well as to establish feasible experimental methods that allow for complete analysis of hard microparticle bulking agents. METHODS: Alumina ceramic beads suspended in hyaluronic acid were injected into the proximal urethra of 15 female rats under an operating microscope. We assessed overall lower urinary tract function, bulking material intraurethral integrity and local host tissue response over time. Microphotographs were taken during injection and again 6 months postoperatively, before urethral harvest. Urinary flow rate and voiding frequency were assessed before and after injection. At 6 months, the urethra was removed and embedded in resin. Hard tissue sections were cut using a sawing microtome, and processed for histological analysis using scanning electron microscopy, light microscopy and immunohistochemistry. RESULTS: Microphotographs of the urethra showed complete volume retention of the bulking agent at 6 months. There was no significant difference between average urinary frequency and mean urinary flow rate at 1 and 3 months postinjection as compared with baseline. Scanning electron microscopy proved suitable for evaluation of microparticle size and integrity, as well as local tissue remodeling. Light microscopy and immunohistochemistry allowed for evaluation of an inflammatory host tissue reaction to the bulking agent. CONCLUSIONS: The microsurgical injection technique, in vivo physiology and novel hard tissue processing for histology, described in the present study, will allow for future comprehensive preclinical testing of urethral bulking therapy agents containing microparticles made of a hard material.

Anatomical and immunohistochemical characteristics of the prostate gland in the greater cane rat (Thryonomys swinderianus).

This study examined the morphology and immunohistochemical features of the prostate gland in 15 captive-reared male greater cane rat of known reproductive and medical history. Samples of the glands were taken after gross examination and routinely prepared for both histological and ultrastructural analysis. Immunohistochemistry was also carried out on paraffin-embedded sections of the glands using rabbit polyclonal antibodies against oestrogen receptors (ERα and ERß) and mouse monoclonal antibody for the progesterone receptor (PR). The prostate, which constitutes 0.04% of the body weight, was a paired, lobulated, brownish gland having three left and four right lobes that partly cover the pelvic urethra. Based on the amount and arrangement of the secretory epithelial folding and relative to their distances to the urethra, two histological zones, the central and peripheral, were identified. However, the epithelium of both zones was lined by predominantly simple cuboidal cells with occasional basal cells. The main ultrastructural features of these cuboidal cells were the presence of several nuclear pores on the nucleus, moderately well-developed, short microvilli and bleb-like apical projections, as well as inter-cellular lacunae seen between these cells and the basal cells. The cuboidal epithelial cells also showed positive nuclear staining for ERα and ERß but not for PR. It is however interesting that the ERα-positive staining was more at the epithelial cells, which is uncommon. These findings highlight the peculiarities in the structure and ultrastructure as well as the unique expression of the oestrogen receptors in the prostate gland of the greater cane rat.

Structure, histochemistry, ultrastructure and seasonal variations of the male prostatic complex in the black Myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae).

Chiroptera are one of the most diverse orders of mammals and a unique group within Mammalia that posses a wide geographic distribution and considerable variability in reproductive strategies. The aims of the present study were to characterise the male prostatic complex of the bat Myotis nigricans (Vespertilionidae) and evaluate seasonal variations in the prostatic complex of M. nigricans specifically. Twenty-three sexually mature specimens (four sample groups: winter, spring, summer and autumn) were subjected to macroscopic, microscopic, morphometric and ultrastructural analyses. The reproductive accessory glands of M. nigricans were found to be composed of a multilobed complex associated with the urethra and a pair of inguinal bulbourethral glands. The complex was composed of three bilobed prostatic regions (ventral, dorsolateral and dorsal) with no ampullary gland and seminal vesicles. This pattern of lobulation is very similar to that described for the prostate of rodents; however, it differs from that of other mammals and even other families of bats (e.g. Phyllostomidae and Molossidae). Each prostatic region in M. nigricans has unique and distinctive characteristics, which synchronise to establish the main reproductive peak of the species in summer. The data also indicated an asynchrony in the activity of primary and secondary reproductive organs in the annual reproductive cycle of M. nigricans in São Paulo State, Brazil.

Most of the patients with suburethral sling failure have tapes located outside the high-pressure zone of the urethra.

OBJECTIVES: The high-pressure zone of the urethra (HPZ), which is crucial for the continence mechanism, extends between the point of the maximum urethral closure pressure and the urethral knee, and has been calculated to lie between 53% and 72% of the functional urethral length. According to recent studies the best results of suburethral slings are achieved when tapes are positioned under this zone. The aim of the study was to determine the location of tapes relative to the urethral length in patients seeking help due to recurrent stress urinary incontinence (SUI) following sling procedures. MATERIAL AND METHODS: The study group comprised 61 patients suffering from recurrent SUI following suburethral slings performed from 6 months to 5 years earlier Forty-nine (80.3%) women were initially treated with a transobturator sling and 12 (19.7%) with a retropubic procedure. Twenty patients had the original sling performed at our department whereas, the other 41 in other institutions. The position of the tapes was determined at the sagittal plane by 3-D transvaginal ultrasound using a linear transducer The length of the urethra was measured from the bladder neck to the external urethral meatus following the urethral lumen, taking into account its curve. The position of the tapes relative to the percentage of the urethral length was calculated assuming the bladder neck as the proximal end of the urethra. The reference point was set at the midpoint on the tape. RESULTS: Only 13 (21.3%) patients had tapes positioned at 50%-75% of the urethral length. In 45 (73.8%) of women examined the tapes were found under proximal half of the urethra and in 3 (4.9%) distally to the 75% of the urethral length. CONCLUSIONS: In most patients in whom slings procedures proved unsuccessful the tapes are located under the proximal half of the urethra, that is outside the HPZ The position of a.tape outside the HPZ may be considered as a cause of suburethral sling failure.

Three-dimensional structure of a parameatal urethral cyst by scanning electron microscopy.

We report scanning electron microscopy (SEM) for a case of parameatal urethral cyst. A 6-year-old Japanese boy presented with a cyst on the right lateral side of the urethral meatus. Histological examination revealed a cyst lined with columnar epithelium. An immunohistochemical study showed positive staining for CK7, CK13, and CEA, and negative for CK20 in luminal cells. On SEM examination, the inner surface of the cyst showed ridges arranged in a gyrus-like manner at lower magnification. Higher magnification revealed luminal cells with short microvillus projections. Some cells showed apocrine, merocrine, and possibly holocrine-type secretions.

Urethral reconstruction using allogenic frozen-thawed bladder mucosa: an experimental study.

OBJECTIVE: To determine the feasibility and effectivity of allogenic frozen-thawed bladder mucosa for urethroplasty. METHODS: Bladder mucosa was harvested from 6 New Zealand rabbits. Changes in the bladder mucosa as seen by histological and electron microscope examination were compared between the frozen-thawed and fresh groups. Twelve urethral stricture models were established and randomly divided into two groups. In the test group, we performed urethroplasty with allogenic frozen-thawed bladder mucosa, and the same operation was done in the control group, but using fresh bladder mucosa. The result of retrograde urethrography and histological changes of the urethral sample were compared postoperatively. RESULTS: No obvious changes on histological and electron microscope examination were observed in the frozen-thawed bladder mucosa. Inflammation reaction of the surgical site in the test group was milder than that of the controls 2 weeks after surgery. The urethral epithelial cells grew well 2 weeks after surgery, but lots of epithelia were necrotic in the control group. The urethra of all rabbits in the test group had good continuity and the urethral lumen was large in the test group 2 months after surgery. There was a layer of urethral epithelium in the test group 2 months after surgery, whereas scar tissue was found in the control group. CONCLUSIONS: The freeze-thaw technique can maintain bladder mucosa structure and biological function. Frozen-thawed allogenic bladder mucosa may be a potential material for urethroplasty.

Regional distribution of neuroendocrine cells in the urogenital duct system of the male rat.

BACKGROUND: Neuroendocrine (NE) cells are frequently present in the human prostate and urethra, whereas they are lacking in the other urogenital organs. This study was undertaken as there are only few detailed studies available on the distribution, form and function of NE cells and the structure of excretory ducts of the accessory sex organs in the male rat. METHODS: Systematic gross anatomical dissections were combined with immunohistochemical and electron microscopic studies of the excretory ducts of the urogenital glands in male rats, with particular focus on the distribution and ultrastructure of the NE cells. RESULTS: The topography and structure of the excretory ducts of the different glands were characterized in detail and analyzed for the distribution of NE cells. These are present (in falling frequencies) in the ducts of seminal vesicles and ventral and lateral prostate and are rare in ducts of coagulating gland, dorsal prostate, urethral epithelium, and excretory ducts of the (bulbo) urethral glands. They are absent in the respective glands proper, the deferent duct and ejaculatory ampulla. Approximately 40% of the NE cells of the ventral prostate ducts are of the "open" type, whereas these are less frequent (14%) in the seminal vesicle ducts, where the "closed" type prevails. CONCLUSIONS: NE cells are present in unequal quantities in the excretory ducts of the accessory sex glands, but they are absent in the glands proper and the deferent ducts. This distribution pattern points to a strictly localized function and differentiation potency of NE precursor cells.

Identification of telocytes in the upper lamina propria of the human urinary tract.

The upper lamina propria (ULP) area of interstitial cells (IC) has been studied extensively in bladder, but is rather unexplored in the rest of the urinary tract. This cell layer is intriguing because of the localization directly underneath the urothelium, the intercellular contacts and the close relationship with nerve endings and capillaries. In this study, we examine the ULP layer of IC in human renal pelvis, ureter and urethra, and we make a comparison with ULP IC in bladder. Tissue was obtained from normal areas in nephrectomy, cystectomy and prostatectomy specimens, and processed for morphology, immunohistochemistry and electron microscopy. A morphological and immunohistochemical phenotype for the ULP IC was assessed and region-dependent differences were looked for. The ULP IC in renal pelvis, ureter and urethra had a similar ultrastructural phenotype, which differed somehow from that of bladder IC, that is, thinner and longer cytoplasmic processes, no peripheral actin filaments and presence of dense core granules and microtubules. Together with their immunohistochemical profile, these features are most compatible with the phenotype of telocytes, a recently discovered group of stromal cells. Based on their global ultrastructural and immunohistochemical phenotype, ULP IC in human bladder should also be classified as telocytes. The most striking immunohistochemical finding was the variable expression of oestrogen receptor (ER) and progesterone receptor (PR). The functional relevance of ULP telocytes in the urinary tract remains to be elucidated, and ER and PR might therefore be promising pharmacological research targets.

Fibroelastic components of the vesicourethral junction of rats in the aging process / Componentes fibroelásticos de la unión vesicouretral de ratas en el proceso de envejecimiento

The vesicourethral junction comprising the vesical trigone, is relevant in setting and positioning of the urinary bladder, along with the vesical neck, fixed by lateral ligaments of the bladder and tendinous arch of the pelvis fascia. Namely,the puboprostatic ligament (men) and the pubovesical (women). The circular set elastic fibers in this junction are important and valuable in the elasticity and plasticity of the area, allowing quick expansion and withdrawal with the flow of urine, and associated to smooth muscle tissue and nerve control form an important collective to maintain urinary continence. The objective of the present study is to describe the elastic system in the vesicouretral junction in relation to aging and its involvement in the states of urinary continence and incontinence, as well as the study of the vesicouretral junction in various age groups while evaluating with electron transmission microscopy. To carry out the study, 12 Wistar rats were used, divided into groups: neonate (4 animals), adult group (4 animals) and aged group (4 animals). Electron transmission microscopy with use of tanic acid technique associated to glutaraldehyde fixation, satisfactorily showed the extreme structural differences between mature elaunin and oxytalan fibers present between intercelular spaces and bundles of collagen fibers. The phases of elastogenesis in neonate animals and degradation of the elastic system of older animals were also evaluated.

La unión vesico-uretral, componente del trígono vesical, posee una relevante importancia en la fijación y posicionamiento de la vejiga urinaria conjuntamente con el cuello vesical, fijado por los ligamentos laterales de la vejiga y arco tendinoso de la fascia de la pelvis. Principalmente, sus componentes anteriores son: el ligamento puboprostático en los hombres y el ligamento pubovesical en las mujeres. Las fibras elásticas dispuestas circularmente en esta unión, son de valiosa importancia en la elasticidad y plasticidad de la región, permitiendo expansión y retiro rápido con el flujo de la orina, y asociado a musculatura lisa y control nervioso forman un conjunto importante para el mantenimiento de la continencia urinaria. Debido a existencia de puntos no esclarecidos en esta región en relación al sistema elástico y su participación en los estados de continencia/incontinencia urinaria, el presente trabajo tiene por objetivo el estudio de la unión vesico-uretral evaluándola en diferentes grupos etarios, a través de la microscopía electrónica de transmisión. Fueron utilizados 12 ratones Wistar, divididos en grupo de neonatos (4 animales), grupo adulto (4 animales) y grupo de ratones viejos (4 animales). La microscopía electrónica de transmisión, con uso de la técnica del ácido tánico asociado al fijador glutaraldeído, mostró satisfactoriamente las diferencias ultraestructurales entre las fibras elásticas maduras, elaunínicas y oxitalánicas, presentes entre los espacios intercelulares de las células musculares y haces de fibras colágenas, y también fases de elastogénesis en animales neonatos y envejecimiento y degradación del sistema elástico en los animales mayores.

Fkbp52 regulates androgen receptor transactivation activity and male urethra morphogenesis.

Hypospadias is a common birth defect in humans, yet its etiology and pattern of onset are largely unknown. Recent studies have shown that male mice with targeted ablation of FK506-binding protein-52 (Fkbp52) develop hypospadias, most likely due to actions of Fkbp52 as a molecular co-chaperone of the androgen receptor (AR). Here, we further dissect the developmental and molecular mechanisms that underlie hypospadias in Fkbp52-deficient mice. Scanning electron microscopy revealed a defect in the elevation of prepucial swelling that led to the onset of the ventral penile cleft. Interestingly, expression of Fkbp52 was highest in the ventral aspect of the developing penis that undergoes fusion of the urethral epithelium. Although in situ hybridization and immunohistochemical analyses suggested that Fkbp52 mutants had a normal urethral epithelium signaling center and epithelial differentiation, a reduced apoptotic cell index at ventral epithelial cells at the site of fusion and a defect of genital mesenchymal cell migration were observed. Supplementation of gestating females with excess testosterone partially rescued the hypospadic phenotype in Fkbp52 mutant males, showing that loss of Fkbp52 desensitizes AR to hormonal activation. Direct measurement of AR activity was performed in mouse embryonic fibroblast cells treated with dihydrotestosterone or synthetic agonist R1881. Reduced AR activity at genes controlling sexual dimorphism and cell growth was found in Fkbp52-deficient mouse embryonic fibroblast cells. However, chromatin immunoprecipitation analysis revealed normal occupancy of AR at gene promoters, suggesting that Fkbp52 exerts downstream effects on the transactivation function of AR. Taken together, our data show Fkbp52 to be an important molecular regulator in the androgen-mediated pathway of urethra morphogenesis.

Effects of birth trauma and estrogen on urethral elastic fibers and elastin expression.

OBJECTIVES: To investigate the effects of birth trauma and estrogen on urethral elastic fibers and elastin expression. METHODS: Pregnant rats were subjected to sham operation (Delivery-only), DVDO (delivery, vaginal distension and ovariectomy), or DVDO + E2 (estrogen). At 2, 4, 8, or 12 weeks, their urethras were harvested for elastic fiber staining and reverse transcription-polymerase chain reaction analysis. Urethral cells were treated with transforming growth factor- ß1 (TGFß1) and/or estrogen and analyzed for elastin mRNA expression. Urethral cells were also examined for the activities of Smad1- and Smad3/4-responsive elements in response to TGFß1 and estrogen. RESULTS: At 8 weeks post-treatment, the urethras of DVDO rats had fewer and shorter elastic fibers when compared with Delivery-only rats, and those of DVDO + E2 rats had fewer and shorter elastic fibers when compared with DVDO rats. Elastin mRNA was expressed at low levels in Delivery-only rats and at increasingly higher levels in DVDO rats at 2, 4, and 8 weeks but at sharply lower levels in DVDO + E2 rats when compared with DVDO rats at 8 weeks. Urethral cells expressed increasingly higher levels of elastin mRNA in response to increasing concentrations of TGFß1 up to 1 ng/mL. At this TGFß1 concentration, urethral cells expressed significantly lower levels of elastin mRNA when treated with estrogen before or after TGFß1 treatment. Both Smad1- and Smad3/4-responsive elements were activated by TGFß1 and such activation was suppressed by estrogen. CONCLUSIONS: Birth trauma appears to activate urethral elastin expression via TGFß1 signaling. Estrogen interferes with this signaling, resulting in improper assembly of elastic fibers.