bromochloroacetic-acid and Hypospadias

Controversy persists regarding the formation of human penile urethra. The classic fusion theory for the development of the spongy urethra and ectodermal ingrowth or endodermal transformation theories for the development of the glanular urethra do not explain the wide spectrum of anomalies seen in patients with hypospadias. This histological study was made to clarify the mechanism of urethral development.. 15 human male embryos ranging from 6 to 14 weeks were studied. The phalluses were examined microscopically and photographed. Tissues were prepared as serial histological sections and stained with haematoxylin and eosin and with special immuno-histochemical stains.. 1) The penile urethra: At 6 weeks of gestation, the urethral plate which is solid distally and partially grooved proximally becomes grooved distally and has fused proximally by 8 weeks. At 14 weeks of gestation; the urethral opening migrates only to the middle of the shaft. 2) The glanular urethra: At the 6th week of gestation, a solid epithelial plate reached the tip of the genital tubercle, and a glans cannot be identified. At the 7th week, a central vacuolation appears and the penile urethral groove does not reach the tip of the phallus. At the 8th week; coronal sulcus starts to appear, and a well defined blind central canal was evident in the 13th week. During the 14th week, the floor of the glanular canal degenerated to form a glanular groove.. Our observations suggest that the spongy urethra passes through 3 stages of development: a solid epithelial plate, deep urethral groove, and fused urethra. The glanular urethra passes through 4 developmental stages: a solid epithelial plate, a blind central canal, a deep glanular groove, and the floor from the preputial lamella. There was no evidence of ectodermal ingrowth. These observations raise serious questions to the current theories for human urethra development. Further studies on fresh human embryos are needed.

Topics: Fetal Development; Gestational Age; Humans; Hypospadias; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Urethra; Vimentin

To study the etiology of hypospadias, we propose the use of a mouse model, the embryonic mouse genital tubercle. In this study, we define the development of the mouse genital tubercle with special emphasis on urethral formation demonstrating anatomical similarities to human development.. Serial sections of genital tubercles from embryonic male and female mice ages 14 to 21 days gestation from timed pregnant animals, newborn and adult mice were immunohistochemical stained with antibodies to E-cadherin, cytokeratins 7, 10, and 14. Patency of the urethral was assessed by india ink injection via the bladder. Urethral lumen morphology was determined by the creation of plastic resin cast. Surface morphology of the genital tubercle was defined by scanning electron microscopy.. India Ink injection into the bladder showed that the urethral lumen was patent from 14 days gestation. Plastic resin casts revealed that the male urethra was characterized by a S shaped curve, the presence of the bulbar urethral gland and a longer length than age matched females. The ontogeny of the genital tubercle development revealed two epithelial edges that subsequently touched and fused into the completed urethra. During development cytokeratin immunohistochemical staining demonstrated that the epithelial cells of the urethral lumen are of bladder origin and the surface cells of skin origin.. The functional and developmental anatomy of the mouse genital tubercle provides a useful model to study normal and abnormal human urethral development.

Topics: Animals; Animals, Newborn; Cadherins; Embryonic and Fetal Development; Female; Gestational Age; Humans; Hypospadias; Infant, Newborn; Keratins; Male; Mice; Microscopy, Electron, Scanning; Models, Animal; Urethra; Urinary Bladder

Congenital genital abnormalities have a diverse spectrum from hypospadias to cloacal anomalies. The molecular events in the normal and abnormal development of the genital tubercle (GT) are still obscure. Genetically engineered mice with specific gene deletions that affect genital anatomy are a useful tool to better understand the etiology of genital abnormalities. In this study, we compared the genital tubercle anatomy of the androgen receptor (AR) deficient, fibroblastic growth factor (FGF)-10 deficient and Sonic HedgeHog (Shh) deficient mutant male mice to that of the wild type male and female mouse.. The lower pelvis of the androgen receptor deficient, FGF-10 deficient, Shh deficient mutant male and wild type male and female mouse at different gestational days (E13-21) and post natal ages (1 day-1 week) were studied. GTs were imaged, serially sectioned and stained immunohistochemically with antibodies raised against E-Cadherin, Cytokeratin 7, 10 and 14. Serial sections of the GTs were selected and three-dimensional computerized images were created to better elucidate the anatomy.. AR deficient mutant male mouse revealed a distinctive GT anatomy, different from both sexes. The corporal bodies and glans remained hypoplastic whereas the urethral spongiosa was more developed than the wild type female counterpart. This finding is consistent with the AR mutant mouse being a unique morphologic phenotype distinct from the normal male and female. FGF-10 deficient mutant male mouse revealed normal corporal bodies with failure of the urethral plate to fuse ventrally consistent with hypospadias. The Shh deficient mutant mouse demonstrated complete agenesis of GT outgrowth and a persistent cloaca.. Animal models bred by gene knockout technology or natural occurring mutants contribute to the basic understanding of normal and abnormal GT development. The anatomy of the these three mutant mice confirms the importance of the androgen receptor, FGF-10 and Shh in genital development.

Topics: Aging; Animals; Animals, Newborn; Female; Fibroblast Growth Factor 10; Fibroblast Growth Factors; Gestational Age; Hedgehog Proteins; Humans; Hypospadias; Keratins; Male; Mice; Mice, Knockout; Models, Animal; Mutation; Receptors, Androgen; Trans-Activators

To describe the detailed anatomy of the urethral plate in relation to its controversial role in hypospadias surgery.. A newborn penis with proximal penile hypospadias and two fetal penises with distal shaft hypospadias were included in the study; 30 normal fetal penises served as the control. Specimens were embedded in paraffin and serially sectioned (6 microm) after formalin fixation. Every 10th section was stained with haematoxylin and eosin. Immunohistochemical staining for nerves (S100), smooth muscles (alpha-actin), blood vessels (factor VIII) and epithelium (cytokeratins 7, 14 and 18) were used on selected sections, with particular attention to the urethral plate. Masson's trichrome and Sirius Red stains were used to localize collagen.. There were extensive blood vessels, glands and smooth muscle under the urethral plate in the hypospadias specimens. These relatively well organized tissues corresponded to an abnormally formed corpus spongiosum. The glands underneath the urethral plate and adjacent to the normal urethra showed positive staining for cytokeratins 7 and 18, respectively (markers of endodermal origin) but were negative for cytokeratin 14 (a marker of ectodermal origin). Penile skin and urethral plate epithelium stained positively for cytokeratin 14 but not for cytokeratin 7 and 18. The urethral plate has a rich nerve supply, as determined by S100 staining. Collagen intensity under the urethral plate was no different from that in normal areas. Tunica albuginea stained intensely for type I and III collagen.. These results show that the urethral plate is well vascularized, has a rich nerve supply and an extensive muscular and connective tissue backing. These features may explain the lower complication rate with onlay flaps than with tube flaps. Therefore, from these anatomical findings, we continue to advocate preservation of the urethral plate and the onlay island flap for hypospadias reconstruction.

Topics: Case-Control Studies; Collagen; Embryonic and Fetal Development; Epithelium; Female; Humans; Hypospadias; Immunohistochemistry; Infant, Newborn; Keratins; Male; Muscle, Smooth; Penis; Pregnancy; Pregnancy Trimester, Second; S100 Proteins; Urethra